WO2025166181A1 - Selective delivery of agents to myeloid-derived suppressor cells with a dendrimer-based formulation - Google Patents

Selective delivery of agents to myeloid-derived suppressor cells with a dendrimer-based formulation

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Publication number
WO2025166181A1
WO2025166181A1 PCT/US2025/014062 US2025014062W WO2025166181A1 WO 2025166181 A1 WO2025166181 A1 WO 2025166181A1 US 2025014062 W US2025014062 W US 2025014062W WO 2025166181 A1 WO2025166181 A1 WO 2025166181A1
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optionally substituted
compound
instance
pharmaceutically acceptable
acceptable salt
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French (fr)
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Fan Zhang
Jeffrey Harrison
Chenikkayala Siva SANKARA
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University of Florida
University of Florida Research Foundation Inc
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University of Florida
University of Florida Research Foundation Inc
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
    • A61K47/59—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes
    • A61K47/595—Polyamides, e.g. nylon
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
    • A61K47/6921—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere
    • A61K47/6927—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores
    • A61K47/6929—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores the form being a nanoparticle, e.g. an immuno-nanoparticle
    • A61K47/6931—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores the form being a nanoparticle, e.g. an immuno-nanoparticle the material constituting the nanoparticle being a polymer
    • A61K47/6935—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores the form being a nanoparticle, e.g. an immuno-nanoparticle the material constituting the nanoparticle being a polymer the polymer being obtained otherwise than by reactions involving carbon to carbon unsaturated bonds, e.g. polyesters, polyamides or polyglycerol

Definitions

  • Nanoparticles have been used to deliver therapeutic payload for disease treatment. While most in vivo biodistribution studies of nanoparticles have been focused on their tissue-level accumulation and clearance 1, 2 , recent progress in the nanomedicine field suggested that targeting nanoparticles to immune cells can be used to modulate the immune response and to enhance therapeutic delivery to the disease region 3-5 .
  • M-MDSCs monocytic-myeloid-derived suppressor cells
  • M-MDSCs are important cellular targets in cancer 6 .
  • M-MDSCs are pathologically activated monocytes with potent immunosuppressive activities. Clinically, a high burden of M-MDSCs is associated with poor prognosis of many solid tumors 7 .
  • M-MDSCs help create and maintain an immunosuppressive tumor microenvironment (TME) 8, 9 . These cells can suppress anti-tumor T cells and promote regulatory T cells and anti-inflammatory myeloid cells 8, 9 . As such, there is an urgent need to develop delivery strategies to target M-MDSCs for cancer treatment. M-MDSCs are also associated with sepsis. [0004] In addition to their immunosuppressive features, M-MDSCs, as phagocytes, could also significantly affect the in vivo fate of nanoparticles. Historic studies have established the roles of myeloid cells in clearing nanoparticles 10, 11 , while emerging evidence has shown that myeloid cells in circulation can take up nanoparticles and actively transport them to the inflamed tissue 4, 12, 13 .
  • M- MDSCs are significantly elevated in the peripheral blood of high-grade glioblastoma patients, accounting for as much as 10% of total cells in the peripheral blood and 30% of total peripheral blood mononuclear cells 14, 15 .
  • the bone marrow accelerates monopoiesis and enhances the egress of M-MDSCs to the systemic circulation, leading to significant expansion of their population in the peripheral blood and in the spleen 16 .
  • Tumor constantly recruits M-MDSCs in large amounts through CCR2-mediated chemotaxis to replenish the tumor-associated macrophages (TAMs) 17, 18 .
  • TAMs tumor-associated macrophages
  • M-MDSCs Because of the abundance of M-MDSCs and their constant infiltration to the tumor sites, M-MDSCs have potential in mediating nanoparticle deposition at the tumors. [0005] Given the roles of M-MDSCs in establishing TME and mediating nanoparticle tumor- targeting, many efforts have tried to establish the correlation between nanoparticle physiochemical properties, such as size, surface charge, and targeting ligand to their targeting of M-MDSCs 19-22 .
  • Dendrimers represent a class of ultra-small nanoparticles with sub-10nm size and carrying drug payload on their surfaces. About 26 dendrimer-based therapeutics with various types of payloads are currently under Phase I-III clinical trials 25 . Systemically administrated hydroxyl-terminated PAMAM dendrimer has been shown to selectively target TAMs in murine glioblastoma (glioma) models 26, 27 . However, given the heterogeneous nature of TAMs, it is unclear what subset(s) are being targeted and what mechanism mediates the selective cell-targeting.
  • STAT3 Signal transducer and activator of transcription 3
  • LLL12 LLL12
  • STAT3 small molecule inhibitor of STAT3
  • the present disclosure provides compounds of Formula (I): and pharmaceutically acceptable salts thereof, wherein L, X, Y, Z, n, p, and q are as defined herein.
  • the present disclosure provides a compound of Formula (I′): and pharmaceutically acceptable salts thereof, wherein L, X, Y, Z, n, p, and q are as defined herein.
  • the present disclosure provides compounds of Formula (II): and pharmaceutically acceptable salts thereof, wherein L, Y, Z n , n, p, and q are as defined herein.
  • the present disclosure provides compounds of Formula (III): and pharmaceutically acceptable salts thereof, wherein L, Y, Z n , n, p, and q are as defined herein.
  • the present disclosure provides compounds of Formula (IV): and pharmaceutically acceptable salts thereof, wherein L, Y, Z n , n, p, and q are as defined herein.
  • the present disclosure provides compounds of Formula (V): and pharmaceutically acceptable salts thereof, wherein L, Y, Z n , n, p, and q are as defined herein. [0015] In another aspect, the present disclosure provides compounds of Formula (VI): and pharmaceutically acceptable salts thereof, wherein L, Y, Z n , n, p, and q are as defined herein.
  • the present disclosure provides compounds of Formula (VII): and pharmaceutically acceptable salts thereof, wherein L, Y, Z n , n, p, and q are as defined herein.
  • the present disclosure provides compounds of Formula (VIII): and pharmaceutically acceptable salts thereof, wherein L, Y, Z n , n, p, and q are as defined herein.
  • the present disclosure provides compounds of Formula (IX): and pharmaceutically acceptable salts thereof, wherein L, Y, Z n , n, p, and q are as defined herein.
  • the present disclosure provides compounds of Formula (X): and pharmaceutically acceptable salts thereof, wherein L, Y, Z n , n, p, and q are as defined herein.
  • the present disclosure provides compounds of Formula (XI): and pharmaceutically acceptable salts thereof, wherein L, Y, Z n , n, p, and q are as defined herein.
  • the present disclosure provides a method of treating or preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formulae (I), (I′), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), or (XI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the present disclosure provides a method of modulating a target protein, the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formulae (I), (I′), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), or (XI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the present disclosure provides kits comprising a provided compound or provided composition and instructions for its use.
  • FIGs.1A-1D show that CCR2 RFP/WT CX3CR1 GFP/WT transgenic mice enable direct surveillance of M-MDSCs in mouse glioma model.
  • FIG.1A shows that in mice with established GL261 tumors, the pie graph shows the flow cytometry measurement of the average percentage of M-MDSCs that reside in the reservoir tissues such as bone marrow, spleen, and peripheral blood leukocytes. At 3-4 weeks after tumor initiation, tissues from 6 tumor-bearing mice were analyzed.
  • FIG.1B shows a gating strategy for the M-MDSCs cells in each tissue: the M-MDSCs are defined as the CCR2+/CX3CR1+ population.
  • FIG.1C shows the gating strategy and the average percentage of major cell subsets in the GL261 tumor stroma.
  • CCR2+/CX3CR1+ cells M-MDSCs; CCR2- /CX3CR1+ cells: likely representing the CNS tissue-resident microglia; CCR2-/CX3CR1 meidum cells: CX3CR1 int , likely represent immune cells infiltrate brain tumor from external sources; CCR2+/CX3CR1- cells: CCR2+, likely represent other infiltrate myeloid cells originated outside of the CNS. CCR2-/CX3CR1- cells: other cells, a collection of tumor cells and other tumor stroma cells. At 3-4 weeks after tumor initiation, tissues from 6 tumor-bearing mice were analyzed.
  • FIG.1D shows a confocal microscopy image of tumor (KR158).
  • FIGs.2A-2H show that tumor M-MDSCs efficiently endocytose dendrimers with high capacity.
  • Cy5-labeled OH dendrimers 50mg/kg were systemically injected into GL261 and KR158 tumor bearing transgenic mice (CCR2 WT/RFP CX3CR1 WT/GFP ) at 3-4 weeks post-implantation; at 24 hours post injection, OH dendrimers uptake in myeloid subsets (indicated by Cy5 Median Fluorescence Intensity (MFI)) and the composition of dendrimer-positive cells (indicated by percentage of dendrimer-positive subset) within the tumor were analyzed through flow cytometry.
  • MFI Cy5 Median Fluorescence Intensity
  • FIG.2A shows the capacity of each cell subsets to uptake OH dendrimers within the GL261 gliomas at 24 hours post-injection. This is indicated by the MFI, which representatively measures the median number of dendrimers deposited per single cell.
  • FIG.2B shows the statistical analysis of FIG.2A.
  • FIG.2C shows a representative density plot of GL261 tumor stromal cells (left panel) and in the same tumor, the density plot of OH dendrimer-positive cells (right panel). M-MDSCs are shown in the box.
  • FIG.2D shows the statistical analysis of the density plot of dendrimer-positive cells in FIG.
  • FIG.2E shows the comparison of the mean, upper and lower quartiles of OH dendrimer deposition in tumor as measured by MFI between GL261 and KR158 gliomas.
  • FIG.2F shows the comparison of OH dendrimer uptake capacity (MFI) in each cell subsets between GL261 and KR158 tumors.
  • 2H shows the correlation analysis between the abundance of cell subset (indicated by the percentage of cell subset within all tumor stroma cells) and the dendrimer deposition within the tumor (indicated by Cy5 MFI of all tumor cells) for both GL261 and KR158 tumors.
  • the Pearson correlation coefficient (R) is based on 95% confidence interval. Weak correlation, R>0.2 or R ⁇ -0.2; strong
  • FIGs.3A-3E show the trafficking kinetics of M-MDSC contributes to the dendrimer accumulation in the tumor.
  • FIG.3A shows a graphical illustration shows the trafficking kinetics of M-MDSCs. M-MDSCs are recruited to the brain tumor from bone marrow (hematopoiesis organ) or spleen (temporary reservoir) through systemic circulation (blood).
  • FIG.3B shows a heatmap that shows the OH dendrimer uptake capacity (indicated by Cy5 MFI) for M-MDSCs in bone marrow, spleen, blood, and tumor in GL261 glioma bearing mice (average MFI from 6 mice).
  • FIG.3C shows a histogram that shows the overall OH dendrimer uptake capacity (indicated by Cy5 MFI) for white blood cells isolated from the peripheral blood of GL261 tumor-bearing mice. Control cells (no dendrimer injection). Cell subsets (lymphocytes, monocytes, granulocytes) were gated based on the scattered plot FSC vs. SSC.
  • FIG.3D shows a comparison of the percentage of dendrimer-positive M- MDSCs between 24 hours vs.72 hours after dendrimer injection. Data was obtained from GL261 tumor-bearing mice.
  • flow cytometry analyses were based on 6 GL261 tumor- bearing mice that received systemic injection of 50 mg/kg OH dendrimers. *p ⁇ 0.05, **p ⁇ 0.01.
  • FIGs.4A-4J show that dendrimer surface chemistry affects their interactions with M-MDSC in vivo.
  • dendrimers with different terminal groups i.e. Succinamic acid: SA, hydroxyl: OH, and amine: NH2
  • SA Succinamic acid
  • hydroxyl OH
  • amine NH2
  • tolerable doses 50mg/kg for SA and OH, 10mg/kg for NH2
  • the bone marrow and tumor were isolated for flow cytometry analysis.
  • FIG.4A shows a comparison of the percentage of dendrimer-positive M-MDSCs (% dendrimer+ M-MDSCs) in the bone marrow of healthy (left) and KR158 tumor-bearing mice (right) between SA, OH, and NH2 dendrimers.
  • the % dendrimer+ M- MDSCs measure dendrimer’s ability to ‘target’ M-MDSCs in the tissue.
  • FIG.4B shows comparison of the percentage of dendrimer-positive M-MDSCs in the KR158 tumor between SA, OH, and NH2 dendrimers.
  • FIG.4C shows representative histograms comparing the Cy5 MFI of dendrimer-positive cells for OH, SA, and NH2 dendrimers in the bone marrow of KR158 tumor-bearing mice (top) and healthy mice (bottom) respectively.
  • FIGs.4D-4E show a comparison of the M-MDSC’s capacity to endocytose SA, OH, and NH2 dendrimers (indicated by MFI).
  • Plot shows M-MDSCs isolated from the bone marrow of healthy mice (left) and KR158 tumor bearing mice (right) (FIG.4D) and the KR158 tumors (FIG.4E).
  • FIG.4F shows a comparison of SA and OH dendrimers for the composition of dendrimer-positive cells within the KR158 tumor.
  • FIG.4G shows the heatmap shows the uptake capacity of SA and OH dendrimers (indicated by Cy5 MFI) by different cell subsets within the GL261 tumor. Data displayed is median.
  • FIGs.4H-4I show statistical analysis comparing the SA and OH dendrimers in terms of their uptake capacity (indicated by Cy5 MFI of dendrimer-positive
  • FIG.4H shows a comparison of SA and OH dendrimers for the composition of dendrimer-positive cells within the GL261 tumor.
  • FIGs.5A-5I show the serum proteins associated with dendrimers dictate the interaction between dendrimers and M-MDSCs.
  • FIG.5A shows a schematic illustration of the generation of M- MDSC from the bone marrow of CCR2 WT/RFP CX3CR1 WT/GFP transgenic mice.
  • bone marrow cells were isolated and were cultured in KR158 conditioned media for 5 days.
  • FIG.5B shows flow cytometry analysis shows that exposure of bone marrow cells to KR158 conditioned media for 5 days enriched the CCR2+/CX3CR1+ cells (M-MDSCs) from less than 10% to approximately 59%.
  • FIG.5C shows the histogram compares the uptake of OH, SA, and NH2 dendrimers (indicated by Cy5 MFI) by ex vivo generated M-MDSCs in the absence of mouse serum.
  • M-MDSCs were incubated with dendrimers at room temperature (RT) for 30 minutes.
  • FIG.5D shows a comparison of ⁇ -potentials (zeta potentials) (mV) of serum proteins (triangle) and NH2, OH, and SA dendrimers before (circle) and after incubation with mouse serum (square). Dendrimers were incubated at 0.86 mg/mL in normal murine serum at 37°C for 30 minutes. ⁇ -potentials (mV) determined using DLS.
  • FIG.5E shows a schematic illustration of the experiment flow that determines the influence of serum proteins on the dendrimer uptake by M-MDSCs.
  • FIG.5F shows the plot based on the experiment flow in FIG.5E, which shows dose-dependent dendrimer uptake (indicated by Cy5 MFI) by M-MDSCs after co- incubation with either PBS (solid line) or mouse serum (dotted line) for NH2 (circle), OH (square), and SA (triangle) dendrimers. Each date point is an average of 3 independent experiment.
  • FIG.5G shows a schematic illustration of the experiment flow that determines how heat-inactivation of serum affect the uptake of NH2, OH, and SA dendrimers by M-MDSCs.
  • FIG.5H shows the plot based on the experiment flow in (FIG.5G), which shows the fold change of dendrimer uptake (x-axis) before and after heat-inactivation of mouse serum as a function of dendrimer dose (y-axis).
  • NH2 circle
  • OH square
  • SA triangle.
  • Negative fold change indicates heat inactivation of serum decreased dendrimer uptake.
  • Positive fold change indicates heat inactivation of serum enhanced dendrimer uptake.
  • Each data point is an average of 3 independent experiment.
  • FIG.5I shows a representative scattered plot of heat inactivation of mouse serum enhanced the uptake of OH dendrimers by M-MDSCs. **p ⁇ 0.01, ***p ⁇ 0.005.
  • FIG.6A shows the capacity of each cell subsets to uptake OH dendrimers within the KR158 gliomas at 24 hours post-injection. This is indicated by the MFI, which representatively measures the median number of dendrimers deposited per single cell.
  • FIG.6B shows the statistical analysis of FIG.6A.
  • FIG.6C shows confocal microscopy image of dendrimer (white) distribution within the tumor (KR158) at 24 hours postinjection. Arrow and star indicate the intracellular localization of OH dendrimer with CCR2+/CX3CR1+ M-MDSCs.
  • FIG.7 shows representative fluorescence image of GL261 and KR158 tumor. Arrow indicates the tumor stroma.
  • FIG.8A shows murine bone marrow sagittal cross section showing OH dendrimer (Cy5) signal 24 hours post 50mg/kg injection of (OH) dendrimer. Image generated by fluorescence microscopy 4X magnification panel and optimized for brightness and contrast using Fiji. Box highlighting image zoom panel. Below showing OH dendrimer in the monocyte rich red marrow of spongy bone (circle).
  • FIG.8B shows murine spleen sagittal cross section showing dendrimer (Cy5) signal 24 hours post 50mg/kg injection of OH dendrimer. Image generated by fluorescence microscopy 10X magnification panel. Box highlighting image zoom panel. Below showing dendrimer deposition in monocyte rich red pulp and exclusion from lymphocyte white pulp zones.
  • the Cy5 labelling of the G6 PAMAM dendrimer was carried out by following previously published work 53 .
  • the amine surface G6 PAMAM dendrimer (50 mg) was dissolved in borate buffer (2 mL, pH 8.5) at room temperature.
  • the hydroxyl surface bi-functional dendrimer and acid surface bi-functional dendrimer were synthesized using literature method 54 .
  • the acid surface bi-functional dendrimer (0.000590 mmol, 1.0 eq, 50 mg) was dissolved in DMSO (2 mL) at room temperature, to this solution was added Cy5 NHS ester (0.0022 mmol, 3.66 eq, 1.72 mg) and DIEA (0.0065 mmol, 11 eq, 1.5 uL).
  • the resulting reaction mixture was allowed to stir overnight and dialyzed against DMSO for 8 h by changing the solvent at least two times, which was further dialyzed against DMF for 12 h by changing the solvent two times.
  • FIGs.9A-9B show flow cytometry analyses of dendrimer uptake in blood leukocytes.
  • FIGs 10A-10B show the number-average mean-based size (FIG.10A) and z-potential (mV) (FIG.10B) of hydroxyl (OH), succinamic acid (SA), and amine (NH2) dendrimers.
  • the size and z- potential were measured by dynamic light scattering (DLS) and electrophoretic light scattering (ELS).
  • FIG.10C shows a schematic illustration of the labeling of Cy5 to NH2 (top), OH (middle), and SA (bottom) dendrimers.
  • FIG.11A shows in vitro evaluation of the dose-dependent toxicity of NH2, OH, and SA dendrimers on primary M-MDSCs by cell-titer blue assay. Positive control: cells treated with 1% Triton-X, Negative control: cell without treatment.
  • FIG.11B shows that at 24 hours after systemic injection, NH2 dendrimers (10mg/kg) co-localized with the endothelial cell (indicated by arrow) in tumor stroma of a KR158 glioma established from CCR2 WT/RFP CX3CR1 WT/GFP transgenic mouse.
  • FIG.12A shows NH2 dendrimer distribution within the KR158 tumor at 24 hours post- injection (10mg/kg), Image based on the widefield fluorescence microscopy (left panel) with zoom (right panel).
  • FIG.12B shows OH dendrimer distribution within the KR158 tumor at 24 hours post- injection (50mg/kg), Image based on widefield fluorescence microscopy (left panel) with zoom (right panel).
  • FIG.12C shows SA dendrimer distribution within the KR158 tumor at 24 hours post-injection (50mg/kg), Image based on widefield fluorescence microscopy (left panel) with zoom (right panel).
  • FIG.12D shows 10X merge confocal of tumor from FIG.12A.
  • FIG.12E shows 10X merge confocal of tumor from tumor shown in FIG.12B.
  • FIG.12F shows 10X merge confocal of tumor from tumor shown in FIG.12C.
  • FIG.12G shows 60X merge confocal for amine dendrimer tumor shown in FIG. 12D.
  • FIG.12H shows 60X merge confocal for amine dendrimer tumor shown in FIG.12E.
  • FIG.12I shows 60X merge confocal for amine dendrimer tumor shown in FIG.12F.
  • FIGs.13A-13B show the comparison of overall tumor depositions of SA and OH dendrimers (indicated by Cy5 MFI) in KR158 tumor (FIG.13A) and GL261 tumor (FIG.13B).
  • FIG.14 shows a schematic of delivery of LLL12 via a dendrimer drug conjugate.
  • FIG.15 shows a schematic of characterization of the dendrimer drug conjugate by dynamic light scattering (DLS).
  • FIG.16 shows a schematic of characterization of drug release quantification using high pressure liquid chromatography (HPLC).
  • FIG.17 shows the zeta potential of PAMAM dendrimers (mV) using electrophoretic light scattering in 10 mM NaCl at 0.03 mg/mL. Positive (NH3) refers to a PAMAM dendrimer with positively charged amine surface groups.
  • FIGs.18A-18H show HPLC studies of G6 PAMAM OH dendrimer at 210 nm (FIG.18A), LLL12 at 250 nm (FIG.18B), LLL12-linker at 250 nm (FIG.18C), dendrimer-LLL12 conjugate 12 at 250 nm (FIG.18D), dendrimer-LLL12 conjugate 17 at 250 nm (FIG.18E), dendrimer-LLL12 conjugate 17 release profile at pH 4.5 at 250 nm (FIG.18F), dendrimer-LLL12 conjugate 17 release profile at 210 nm (FIG.18G), and dendrimer-LLL12 conjugate 17 release profile at pH 7.4 at 250 nm (FIG.18H
  • FIGs.19A-19G show IC50 efficacy and toxicity studies.
  • FIG.19A shows a schematic for the IC50 studies.
  • FIG.19B shows a THP-1 STAT3-Luc IL-6 response curve.
  • FIG.19C shows a dose response
  • FIGs.19D-19E show the LLL12 IC50 mass.
  • FIG.19F shows the IC50 for dendrimer-LLL12 conjugate 17.
  • FIG.19G shows a comparison of toxicity between LLL12 and dendrimer-LLL12 conjugate 17.
  • FIGs.20A-20B show a UV-vis spectrum of LLL12 (FIG.20A) and a calibration curve (FIG. 20B).
  • FIGs.21A-21B show 1 H NMR (FIG.21A) and 13 C NMR (FIG.21B) spectra of 1.
  • FIGs.22A-22B show 1 H NMR (FIG.22A) and 13 C NMR (FIG.22B) spectra of 3.
  • FIGs.23A-23B show 1 H NMR (FIG.23A) and 13 C NMR (FIG.23B) spectra of 4.
  • FIG.24 shows 1 H NMR spectrum of 6.
  • FIG.25 shows 1 H NMR spectrum of 9.
  • FIG.26 shows 1 H NMR spectrum of 10.
  • FIG.27 shows 1 H NMR spectrum of 12.
  • FIGs.28A-28B show 1 H NMR (FIG.28A) and 13 C NMR (FIG.28B) spectra of 14.
  • FIG.29 shows 1 H NMR spectrum of 16.
  • FIG.30 shows 1 H NMR spectrum of 17.
  • FIGs.31A-31B show 1 H NMR (FIG.31A) and 13 C NMR (FIG.31B) spectra of 18.
  • FIGs.32A-32B show 1 H NMR (FIG.32A) and 13 C NMR (FIG.32B) spectra of 20.
  • FIGs.33A-33B show 1 H NMR spectra of 21 from 0 ppm to 14 ppm (FIG.33A) and 0 ppm to 9 ppm (FIG.32B).
  • FIGs.34A-34B show (FIG.34A) and Zeta Potential (FIG.34B) of dendrimer conjugates of the present disclosure, along with unfunctionalized G6-OH. Size and Zeta Potential were calculated in 10mM NaCl.
  • FIG.35 shows a schematic of efficacy (IC50) and toxicity analyses for dendrimer-drug conjugates.
  • FIGs.36A-36B show representative IC50 curves for free drug (LLL12), dendrimer-LLL12 conjugates D-LLL12C (17), D-LLL12S (12), and D- LLL12H (21) (FIG.36A) and average IC50 values from 3 repeated experiments (FIG.36B).
  • FIGs.37A-37C show toxicity (measured by cell viability) for 17 (FIG.37A), 12 (FIG.37B), and 21 (FIG.37C), with LLL12 as a control.
  • FIGs.38A-38E show the therapeutic window.
  • FIGs.38A-38D show the overlays of the IC50 curve (bioluminescence; curve with open circles) and the viability curve (curve with shaded triangles) for LLL12 (FIG.38A), G6-LLL12C (D-LLL12C, 17) (FIG.38B), G6-LLL12S (D-LLL12S, 12) (FIG.38C), and G6-LLL12H (D-LLL12H, 21) (FIG.38D).
  • FIG.38E shows the calculated therapeutic windows.
  • Compounds described herein can comprise one or more asymmetric centers, and thus can exist in various stereoisomeric forms, e.g., enantiomers and/or diastereomers.
  • the compounds described herein can be in the form of an individual enantiomer, diastereomer or geometric isomer, or can be in the form of a mixture of stereoisomers, including racemic mixtures and mixtures enriched in one or more stereoisomer.
  • Isomers can be isolated from mixtures by methods known to those skilled in the art, including chiral high pressure liquid chromatography (HPLC) and the formation and crystallization of chiral salts; or preferred isomers can be prepared by asymmetric syntheses. See, for example, Jacques et al., Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981); Wilen et al., Tetrahedron 33:2725 (1977); Eliel, E.L. Stereochemistry of Carbon Compounds (McGraw–Hill, NY, 1962); and Wilen, S.H., Tables of Resolving Agents and Optical Resolutions p.268 (E.L. Eliel, Ed., Univ.
  • range is inclusive of the values at the two ends of the range unless otherwise provided.
  • C1-6 alkyl encompasses, C1, C2, C3, C4, C5, C6, C1–6, C1–5, C1–4, C1–3, C1–2, C2–6, C2–5, C2– 4 , C 2–3 , C 3–6 , C 3–5 , C 3–4 , C 4–6 , C 4–5 , and C 5–6 alkyl.
  • alkyl refers to a radical of a straight-chain or branched saturated hydrocarbon group having from 1 to 20 carbon atoms (“C 1–20 alkyl”). In some embodiments, an alkyl group has 1 to 12 carbon atoms (“C 1–12 alkyl”).
  • an alkyl group has 1 to 10 carbon atoms (“C 1–10 alkyl”). In some embodiments, an alkyl group has 1 to 9 carbon atoms (“C 1–9 alkyl”). In some embodiments, an alkyl group has 1 to 8 carbon atoms (“C 1–8 alkyl”). In some embodiments, an alkyl group has 1 to 7 carbon atoms (“C 1–7 alkyl”). In some embodiments, an alkyl group has 1 to 6 carbon atoms (“C 1–6 alkyl”). In some embodiments, an alkyl group has 1 to 5 carbon atoms (“C 1–5 alkyl”). In some embodiments, an alkyl group has 1 to 4 carbon atoms (“C 1–4 alkyl”). In some embodiments, an alkyl group has 1 to 3 carbon atoms (“C 1–3 alkyl”). In some embodiments, an alkyl group has 1 to 2
  • C 1–2 alkyl U1197.70243WO00 12/193 #13587456v2 carbon atoms (“C 1–2 alkyl”).
  • an alkyl group has 1 carbon atom (“C 1 alkyl”).
  • an alkyl group has 2 to 6 carbon atoms (“C 2-6 alkyl”).
  • C 1–6 alkyl groups include methyl (C 1 ), ethyl (C 2 ), propyl (C 3 ) (e.g., n-propyl, isopropyl), butyl (C 4 ) (e.g., n-butyl, tert-butyl, sec-butyl, isobutyl), pentyl (C 5 ) (e.g., n-pentyl, 3-pentanyl, amyl, neopentyl, 3-methyl-2- butanyl, tert-amyl), and hexyl (C6) (e.g., n-hexyl).
  • alkyl groups include n- heptyl (C 7 ), n-octyl (C 8 ), n-dodecyl (C 12 ), and the like. Unless otherwise specified, each instance of an alkyl group is independently unsubstituted (an “unsubstituted alkyl”) or substituted (a “substituted alkyl”) with one or more substituents (e.g., halogen, such as F).
  • substituents e.g., halogen, such as F
  • the alkyl group is an unsubstituted C1–12 alkyl (such as unsubstituted C1–6 alkyl, e.g., ⁇ 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 or s-Bu), unsubstituted isobutyl (i-Bu)).
  • unsubstituted C1–12 alkyl such as unsubstituted C1–6 alkyl, e.g.
  • the alkyl group is a substituted C1–12 alkyl (such as substituted C1–6 alkyl, e.g., –CH2F, –CHF2, –CF3, – CH2CH2F, –CH2CHF2, –CH2CF3, or benzyl (Bn)).
  • haloalkyl is a substituted alkyl group, wherein one or more of the hydrogen atoms are independently replaced by a halogen, e.g., fluoro, bromo, chloro, or iodo.
  • Perhaloalkyl is a subset of haloalkyl, and refers to an alkyl group wherein all of the hydrogen atoms are independently replaced by a halogen, e.g., fluoro, bromo, chloro, or iodo.
  • halogen e.g., fluoro, bromo, chloro, or iodo.
  • heteroalkyl refers to an alkyl group, which further includes at least one heteroatom (e.g., 1, 2, 3, or 4 heteroatoms) selected from oxygen, nitrogen, or sulfur within (e.g., inserted between adjacent carbon atoms of) and/or placed at one or more terminal position(s) of the parent chain.
  • a heteroalkyl group refers to a saturated group having from 1 to 20 carbon atoms and 1 or more heteroatoms within the parent chain (“C1–20 heteroalkyl”). In certain embodiments, a heteroalkyl group refers to a saturated group having from 1 to 12 carbon atoms and 1 or more heteroatoms within the parent chain (“C1–12 heteroalkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 11 carbon atoms and 1 or more heteroatoms within the parent chain (“C1–11 heteroalkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 10 carbon atoms and 1 or more heteroatoms within the parent chain (“C 1–10 heteroalkyl”).
  • a heteroalkyl group is a saturated group having 1 to 9 carbon atoms and 1 or more heteroatoms within the parent chain (“C 1–9 heteroalkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 8 carbon atoms and 1 or more heteroatoms within the parent chain (“C 1–8 heteroalkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 7 carbon atoms and 1 or more heteroatoms within the parent chain (“C 1–7 heteroalkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 6 carbon atoms and 1 or more heteroatoms within the parent chain (“C 1–6 heteroalkyl”).
  • a heteroalkyl group is a saturated group having 1 to 5 carbon atoms and 1 or 2 heteroatoms within the parent chain (“C 1–5 heteroalkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 4 carbon
  • a heteroalkyl group is a saturated group having 1 to 3 carbon atoms and 1 heteroatom within the parent chain (“C 1–3 heteroalkyl”).
  • a heteroalkyl group is a saturated group having 1 to 2 carbon atoms and 1 heteroatom within the parent chain (“C 1–2 heteroalkyl”).
  • a heteroalkyl group is a saturated group having 1 carbon atom and 1 heteroatom (“C1 heteroalkyl”).
  • a heteroalkyl group is a saturated group having 2 to 6 carbon atoms and 1 or 2 heteroatoms within the parent chain (“C 2-6 heteroalkyl”). Unless otherwise specified, each instance of a heteroalkyl group is independently unsubstituted (an “unsubstituted heteroalkyl”) or substituted (a “substituted heteroalkyl”) with one or more substituents. In certain embodiments, the heteroalkyl group is an unsubstituted C1–12 heteroalkyl. In certain embodiments, the heteroalkyl group is a substituted C1–12 heteroalkyl.
  • alkenyl refers to a radical of a straight-chain or branched hydrocarbon group having from 1 to 20 carbon atoms and one or more carbon-carbon double bonds (e.g., 1, 2, 3, or 4 double bonds).
  • an alkenyl group has 1 to 20 carbon atoms (“C1-20 alkenyl”).
  • an alkenyl group has 1 to 12 carbon atoms (“C1–12 alkenyl”).
  • an alkenyl group has 1 to 11 carbon atoms (“C1–11 alkenyl”).
  • an alkenyl group has 1 to 10 carbon atoms (“C1–10 alkenyl”).
  • an alkenyl group has 1 to 9 carbon atoms (“C1–9 alkenyl”). In some embodiments, an alkenyl group has 1 to 8 carbon atoms (“C1–8 alkenyl”). In some embodiments, an alkenyl group has 1 to 7 carbon atoms (“C1–7 alkenyl”). In some embodiments, an alkenyl group has 1 to 6 carbon atoms (“C1–6 alkenyl”). In some embodiments, an alkenyl group has 1 to 5 carbon atoms (“C1–5 alkenyl”). In some embodiments, an alkenyl group has 1 to 4 carbon atoms (“C1–4 alkenyl”).
  • an alkenyl group has 1 to 3 carbon atoms (“C1–3 alkenyl”). In some embodiments, an alkenyl group has 1 to 2 carbon atoms (“C1–2 alkenyl”). In some embodiments, an alkenyl group has 1 carbon atom (“C1 alkenyl”).
  • the one or more carbon-carbon double bonds can be internal (such as in 2-butenyl) or terminal (such as in 1-butenyl).
  • Examples of C1–4 alkenyl groups include methylidenyl (C1), ethenyl (C2), 1-propenyl (C3), 2-propenyl (C3), 1-butenyl (C4), 2-butenyl (C4), butadienyl (C4), and the like.
  • Examples of C1–6 alkenyl groups include the aforementioned C 2-4 alkenyl groups as well as pentenyl (C 5 ), pentadienyl (C 5 ), hexenyl (C 6 ), and the like. Additional examples of alkenyl include heptenyl (C 7 ), octenyl (C 8 ), octatrienyl (C 8 ), and the like.
  • each instance of an alkenyl group is independently unsubstituted (an “unsubstituted alkenyl”) or substituted (a “substituted alkenyl”) with one or more substituents.
  • the alkenyl group is an unsubstituted C 1-20 alkenyl.
  • the alkenyl group is a substituted C 1-20 alkenyl.
  • heteroalkenyl refers to an alkenyl group, which further includes at least one heteroatom (e.g., 1, 2, 3, or 4 heteroatoms) selected from oxygen, nitrogen, or sulfur within (e.g., inserted between adjacent carbon atoms of) and/or placed at one or more terminal position(s) of the parent chain.
  • heteroatom e.g., 1, 2, 3, or 4 heteroatoms
  • alkynyl refers to a radical of a straight-chain or branched hydrocarbon group having from 1 to 20 carbon atoms and one or more carbon-carbon triple bonds (e.g., 1, 2, 3, or 4 triple bonds) (“C 1-20 alkynyl”).
  • heteroalkynyl refers to an alkynyl group, which further includes at least one heteroatom (e.g., 1, 2, 3, or 4 heteroatoms) selected from oxygen, nitrogen, or sulfur within (e.g., inserted between adjacent carbon atoms of) and/or placed at one or more terminal position(s) of the parent chain.
  • heteroatom e.g., 1, 2, 3, or 4 heteroatoms
  • carbocyclyl or “carbocyclic” refers to a radical of a non-aromatic cyclic hydrocarbon group having from 3 to 14 ring carbon atoms (“C3-14 carbocyclyl”) and zero heteroatoms in the non-aromatic ring system.
  • heterocyclyl refers to a radical of a 3- to 14-membered non- aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“3–14 membered heterocyclyl”).
  • 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 ⁇ electrons shared in a cyclic array) having 6–14 ring carbon atoms and zero heteroatoms provided in the aromatic ring system (“C6-14 aryl”).
  • heteroaryl refers to a radical of a 5-14 membered monocyclic or polycyclic (e.g., bicyclic, tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10, or 14 ⁇ electrons shared in a cyclic array) having ring carbon atoms and 1–4 ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-14 membered heteroaryl”).
  • unsaturated bond refers to a double or triple bond.
  • the term “unsaturated” or “partially unsaturated” refers to a moiety that includes at least one double or triple bond.
  • saturated or “fully saturated” refers to a moiety that does not contain a double or triple bond, e.g., the moiety only contains single bonds.
  • Affixing the suffix “-ene” to a group indicates the group is a divalent moiety, e.g., alkylene is the divalent moiety of alkyl, alkenylene is the divalent moiety of alkenyl, heteroalkylene is the divalent moiety of heteroalkyl, and heteroalkenylene is the divalent moiety of heteroalkenyl.
  • a group is optionally substituted unless expressly provided otherwise.
  • the term “optionally substituted” refers to being substituted or unsubstituted.
  • alkyl, alkenyl, heteroalkyl, and heteroalkenyl groups are optionally substituted.
  • “Optionally substituted” refers to a group which is substituted or unsubstituted (e.g., “substituted” or “unsubstituted” alkyl, “substituted”
  • a “substituted” group has a substituent at one or more substitutable positions of the group, and when more than one position in any given structure is substituted, the substituent is either the same or different at each position.
  • substituted is contemplated to include substitution with all permissible substituents of organic compounds, and includes any of the substituents described herein that results in the formation of a stable compound.
  • the present disclosure contemplates any and all such combinations in order to arrive at a stable compound.
  • heteroatoms such as nitrogen may have hydrogen substituents and/or any suitable substituent as described herein which satisfy the valencies of the heteroatoms and results in the formation of a stable moiety.
  • each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 R gg groups; each instance of R ff is, independently, selected from hydrogen, C 1–10 alkyl, C 1–10 perhaloalkyl, C1–10 alkenyl, C1–10 alkynyl, heteroC1–10 alkyl, heteroC1–10 alkenyl, heteroC1–10 alkynyl, C 3-10 carbocyclyl, 3-10 membered heterocyclyl, C 6-10 aryl, and 5-10 membered heteroaryl, or two R ff groups are joined to form a 3-10 membered heterocyclyl or 5-10 membered heteroaryl ring, wherein each alkyl, alkenyl, alkyl, alkynyl, heteroalkyl, heteroalkeny
  • halo or halogen refers to fluorine (fluoro, ⁇ F), chlorine (chloro, ⁇ Cl), bromine (bromo, ⁇ Br), or iodine (iodo, ⁇ I).
  • hydroxyl or “hydroxy” refers to the group ⁇ OH.
  • Nitrogen atoms can be substituted or unsubstituted as valency permits, and include primary, secondary, tertiary, and quaternary nitrogen atoms.
  • the substituent present on the nitrogen atom is a nitrogen protecting group (also referred to herein as an “amino protecting group”).
  • each oxygen atom substituent is independently substituted (e.g., substituted with one or more halogen) or unsubstituted C 1-6 alkyl or an oxygen protecting group.
  • the substituent present on an oxygen atom is an oxygen protecting group (also referred to herein as an “hydroxyl protecting group”).
  • Oxygen protecting groups include
  • a “counterion” or “anionic counterion” is a negatively charged group associated with a positively charged group in order to maintain electronic neutrality.
  • An anionic counterion may be monovalent (e.g., including one formal negative charge).
  • An anionic counterion may also be multivalent (e.g., including more than one formal negative charge), such as divalent or trivalent.
  • Exemplary counterions include halide ions (e.g., F – , Cl – , Br – , I – ), NO3 – , ClO4 – , OH – , H2PO4 – , HCO3 ⁇ , HSO4 – , sulfonate ions (e.g., methansulfonate, trifluoromethanesulfonate, p–toluenesulfonate, benzenesulfonate, 10–camphor sulfonate, naphthalene–2–sulfonate, naphthalene–1–sulfonic acid–5– sulfonate, ethan–1–sulfonic acid–2–sulfonate, and the like), carboxylate ions (e.g., acetate, propanoate, benzoate, glycerate, lactate, tartrate, glycolate, gluconate, and
  • Exemplary counterions which may be multivalent include CO3 2 ⁇ , HPO4 2 ⁇ , PO4 3 ⁇ , B4O7 2 ⁇ , SO4 2 ⁇ , S2O3 2 ⁇ , carboxylate anions (e.g., tartrate, citrate, fumarate, maleate, malate, malonate, gluconate, succinate, glutarate, adipate, pimelate, suberate, azelate, sebacate, salicylate, phthalates, aspartate, glutamate, and the like), and carboranes.
  • carboxylate anions e.g., tartrate, citrate, fumarate, maleate, malate, malonate, gluconate, succinate, glutarate, adipate, pimelate, suberate, azelate, sebacate, salicylate, phthalates, aspartate, glutamate, and the like
  • carboranes e.g., tartrate, citrate, fumarate, maleate, mal
  • a “leaving group” is an art-understood term referring to an atomic or molecular fragment that departs with a pair of electrons in heterolytic bond cleavage, wherein the molecular fragment is an anion or neutral molecule.
  • a leaving group can be an atom or a group capable of being displaced by a nucleophile. See e.g., Smith, March Advanced Organic Chemistry 6th ed. (501–502). In some embodiments, the leaving group is an internal leaving group such as an epoxide.
  • a “non-hydrogen group” refers to any group that is defined for a particular variable that is not hydrogen.
  • heteroatom refers to an atom that is not hydrogen or carbon. In certain embodiments, the heteroatom is nitrogen. In certain embodiments, the heteroatom is oxygen. In certain embodiments, the heteroatom is sulfur.
  • salt refers to any and all salts, and encompasses pharmaceutically acceptable salts. Salts include ionic compounds that result from the neutralization reaction of an acid and a base. A salt is composed of one or more cations (positively charged ions) and one or more anions (negative ions) so that the salt is electrically neutral (without a net charge). Salts of the compounds of the present disclosure include those derived from inorganic and organic acids and bases.
  • acid addition salts are salts of an amino group formed with inorganic acids, such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid, or with organic acids, such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid or by using other methods known in the art such as ion exchange.
  • inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid
  • organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid or by using other methods known in the art such as ion exchange.
  • salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2–hydroxy–ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2–naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, per
  • Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N + (C1–4 alkyl)4 salts.
  • Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like.
  • Further salts include ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, lower alkyl sulfonate, and aryl sulfonate.
  • pharmaceutically acceptable salt refers to those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response, and the like, and are commensurate with a reasonable benefit/risk ratio.
  • Pharmaceutically acceptable salts are well known in the art. For example, Berge et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19, incorporated herein by reference.
  • Pharmaceutically acceptable salts of the compounds of the present disclosure include those derived from suitable inorganic and organic acids and bases.
  • Examples of pharmaceutically acceptable, nontoxic acid addition salts are salts of an amino group formed with inorganic acids, such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid or with organic acids, such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid or by using other methods known in the art such as ion exchange.
  • inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid
  • organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid or by using other methods known in the art such as ion exchange.
  • salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2- hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate,
  • Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium, and N + (C 1-4 alkyl)4 salts.
  • Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like.
  • Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, lower alkyl sulfonate, and aryl sulfonate.
  • An enantiomer can be characterized by the absolute configuration of its asymmetric center and is described by the R- and S-sequencing rules of Cahn and Prelog, or by the manner in which the molecule rotates the plane of polarized light and designated as dextrorotatory or levorotatory (i.e., as (+) or ( ⁇ )-isomers respectively).
  • a chiral compound can exist as either individual enantiomer or as a mixture thereof.
  • a mixture containing equal proportions of the enantiomers is called a “racemic mixture”.
  • a “subject” to which administration is contemplated refers to a human (i.e., male or female of any age group, e.g., pediatric subject (e.g., infant, child, or adolescent) or adult subject (e.g., young adult, middle-aged adult, or senior adult)) or non-human animal.
  • the non- human animal is a mammal (e.g., primate (e.g., cynomolgus monkey or rhesus monkey), commercially relevant mammal (e.g., cattle, pig, horse, sheep, goat, cat, or dog), or bird (e.g., commercially relevant bird, such as chicken, duck, goose, or turkey)).
  • primate e.g., cynomolgus monkey or rhesus monkey
  • commercially relevant mammal e.g., cattle, pig, horse, sheep, goat, cat, or dog
  • bird e.g., commercially relevant bird, such as
  • the non-human animal is a fish, reptile, or amphibian.
  • the non-human animal may be a male or female at any stage of development.
  • the non-human animal may be a transgenic animal or genetically engineered animal.
  • patient refers to a human subject in need of treatment of a disease.
  • tissue samples such as tissue sections and needle biopsies of a tissue
  • cell samples e.g., cytological smears (such as Pap or blood smears) or samples of cells obtained by microdissection); samples of whole organisms (such as samples of yeasts or bacteria); or cell fractions, fragments or organelles (such as obtained by lysing cells and separating the components thereof by centrifugation or otherwise).
  • Other examples of biological samples include blood, serum, urine, semen, fecal matter, cerebrospinal fluid, interstitial
  • target tissue refers to any biological tissue of a subject (including a group of cells, a body part, or an organ) or a part thereof, including blood and/or lymph vessels, which is the object to which a compound, particle, and/or composition of the present disclosure is delivered.
  • a target tissue may be an abnormal or unhealthy tissue, which may need to be treated.
  • a target tissue may also be a normal or healthy tissue that is under a higher than normal risk of becoming abnormal or unhealthy, which may need to be prevented.
  • the target tissue is the liver.
  • the target tissue is the lung.
  • a “non-target tissue” is any biological tissue of a subject (including a group of cells, a body part, or an organ) or a part thereof, including blood and/or lymph vessels, which is not a target tissue.
  • administer refers to implanting, absorbing, ingesting, injecting, inhaling, or otherwise introducing a compound described herein, or a composition thereof, in or on a subject.
  • treatment refers to reversing, alleviating, delaying the onset of, or inhibiting the progress of a disease described herein.
  • treatment may be administered after one or more signs or symptoms of the disease have developed or have been observed. In other embodiments, treatment may be administered in the absence of signs or symptoms of the disease.
  • treatment may be administered to a susceptible subject prior to the onset of symptoms (e.g., in light of a history of symptoms and/or in light of exposure to a pathogen). Treatment may also be continued after symptoms have resolved, for example, to delay or prevent recurrence.
  • condition e.g., in light of a history of symptoms and/or in light of exposure to a pathogen.
  • Treatment may also be continued after symptoms have resolved, for example, to delay or prevent recurrence.
  • An “effective amount” of a compound described herein refers to an amount sufficient to elicit the desired biological response.
  • an effective amount of a compound described herein may vary depending on such factors as the desired biological endpoint, severity of side effects, disease, or disorder, the identity, pharmacokinetics, and pharmacodynamics of the particular compound, the condition being treated, the mode, route, and desired or required frequency of administration, the species, age and health or general condition of the subject.
  • an effective amount is a therapeutically effective amount.
  • an effective amount is a prophylactic treatment.
  • an effective amount is the amount of a compound described herein in a single dose.
  • an effective amount is the combined amounts of a compound described herein in multiple doses.
  • the desired dosage is delivered three times a day, two times a day, once a day, every other day, every third day, every week, every two weeks, every three weeks, or every four weeks.
  • the desired dosage is delivered three times a day, two times a day, once a day, every other day, every third day, every week, every two weeks, every three weeks, or every four weeks.
  • the desired dosage is
  • U1197.70243WO00 23/193 #13587456v2 desired dosage is delivered using multiple administrations (e.g., two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, or more administrations).
  • an effective amount of a compound for administration one or more times a day to a 70 kg adult human comprises about 0.0001 mg to about 3000 mg, about 0.0001 mg to about 2000 mg, about 0.0001 mg to about 1000 mg, about 0.001 mg to about 1000 mg, about 0.01 mg to about 1000 mg, about 0.1 mg to about 1000 mg, about 1 mg to about 1000 mg, about 1 mg to about 100 mg, about 10 mg to about 1000 mg, or about 100 mg to about 1000 mg, of a compound per unit dosage form.
  • the compounds of the present disclosure are administered orally or parenterally at dosage levels sufficient to deliver from about 0.001 mg/kg to about 100 mg/kg, from about 0.01 mg/kg to about 50 mg/kg, preferably from about 0.1 mg/kg to about 40 mg/kg, preferably from about 0.5 mg/kg to about 30 mg/kg, from about 0.01 mg/kg to about 10 mg/kg, from about 0.1 mg/kg to about 10 mg/kg, and more preferably from about 1 mg/kg to about 25 mg/kg, of subject body weight per day, one or more times a day, to obtain the desired therapeutic effect.
  • dose ranges as described herein provide guidance for the administration of provided pharmaceutical compositions to an adult.
  • a “therapeutically effective amount” of a compound described herein is an amount sufficient to provide a therapeutic benefit in the treatment of a condition or to delay or minimize one or more symptoms associated with the condition.
  • a therapeutically effective amount of a compound means an amount of therapeutic agent, alone or in combination with other therapies, which provides a therapeutic benefit in the treatment of the condition.
  • the term “therapeutically effective amount” can encompass an amount that improves overall therapy, reduces or avoids symptoms, signs, or causes of the condition, and/or enhances the therapeutic efficacy of another therapeutic agent.
  • a therapeutically effective amount is an amount sufficient for modulating a target protein in a subject or a cell, tissue, or biological sample. In certain embodiments, a therapeutically effective amount is an amount sufficient for treating cancer (e.g., a solid tumor (e.g., brain tumor (e.g., glioblastoma), pancreatic cancer, ovarian cancer)). In certain embodiments, a therapeutically effective amount is an amount sufficient for modulating a target protein in a subject or a cell, tissue, or biological sample and treating cancer (e.g., a solid tumor (e.g., brain tumor (e.g., glioblastoma), pancreatic cancer, ovarian cancer)).
  • cancer e.g., a solid tumor (e.g., brain tumor (e.g., glioblastoma), pancreatic cancer, ovarian cancer)).
  • a “prophylactically effective amount” of a compound described herein is an amount sufficient to prevent a condition, or one or more symptoms associated with the condition or prevent its recurrence.
  • a prophylactically effective amount of a compound means an amount of a therapeutic agent, alone or in combination with other agents, which provides a prophylactic benefit in the prevention of the condition.
  • the term “prophylactically effective amount” can encompass an amount
  • a prophylactically effective amount is an amount sufficient for modulating a target protein in a subject or a cell, tissue, or biological sample.
  • a prophylactically effective amount is an amount sufficient for treating cancer (e.g., a solid tumor (e.g., brain tumor (e.g., glioblastoma), pancreatic cancer, ovarian cancer)).
  • a prophylactically effective amount is an amount sufficient for modulating a target protein in a subject or a cell, tissue, or biological sample and treating cancer (e.g., a solid tumor (e.g., brain tumor (e.g., glioblastoma), pancreatic cancer, ovarian cancer)).
  • cancer e.g., a solid tumor (e.g., brain tumor (e.g., glioblastoma), pancreatic cancer, ovarian cancer)
  • prevent refers to a prophylactic treatment of a subject who is not and was not with a disease but is at risk of developing the disease or who was with a disease, is not with the disease, but is at risk of regression of the disease.
  • the subject is at a higher risk of developing the disease or at a higher risk of regression of the disease than an average healthy member of a population.
  • the term refers to a reduction of the level of enzyme activity, e.g., STAT3, IDO, NOS, arginase, PD-L1, protein kinase B, or CD73 activity, to a level that is statistically significantly lower than an initial level, which may, for example, be a baseline level of enzyme activity.
  • a level of enzyme activity e.g., STAT3, IDO, NOS, arginase, PD-L1, protein kinase B, or CD73 activity
  • the term refers to a reduction of the level of enzyme activity, e.g., STAT3, IDO, NOS, arginase, PD-L1, protein kinase B, or CD73 activity, to a level that is less than 75%, less than 50%, less than 40%, less than 30%, less than 25%, less than 20%, less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, less than 0.01%, less than 0.001%, or less than 0.0001% of an initial level, which may, for example, be a baseline level of enzyme activity.
  • the level of enzyme activity e.g., STAT3, IDO, NOS, arginase, PD-L1, protein kinase B, or CD73 activity
  • agonist or “agonism” in the context of enzymes, for example, in the context of TLR7 or STING, refers to the activation of a receptor to produce an enzyme response.
  • the term refers to an increase of the level of enzyme response, e.g., TLR7 or STING response, to a level that is statistically significantly higher than an initial level, which may, for example, be a baseline level of enzyme response.
  • the term refers to an increase of the level of enzyme response, e.g., TLR7 or STING response, to a level that is greater than 100%, greater than 125%, greater than 150%, greater than 175%, greater than 200%, greater than 300%, greater than 400%, greater than 500%, greater than 600%, greater than 700%, greater than 800%, greater than 900%, or greater than 1000% of an initial level, which may, for example, be a baseline level of enzyme response.
  • the term “antagonist” or “antagonism” in the context of enzymes for example, refers to blocking activation of a receptor to prevent an enzyme response. In some embodiments, the term refers to a reduction of the level of enzyme response, to a level that is statistically significantly lower
  • U1197.70243WO00 25/193 #13587456v2 than an initial level which may, for example, be a baseline level of enzyme response.
  • the term refers to a reduction of the level of enzyme response, to a level that is less than 75%, less than 50%, less than 40%, less than 30%, less than 25%, less than 20%, less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, less than 0.01%, less than 0.001%, or less than 0.0001% of an initial level, which may, for example, be a baseline level of enzyme response.
  • a proliferative disease refers to a disease that occurs due to abnormal growth or extension by the multiplication of cells (Walker, Cambridge Dictionary of Biology; Cambridge University Press: Cambridge, UK, 1990).
  • a proliferative disease may be associated with: 1) the pathological proliferation of normally quiescent cells; 2) the pathological migration of cells from their normal location (e.g., metastasis of neoplastic cells); 3) the pathological expression of proteolytic enzymes such as the matrix metalloproteinases (e.g., collagenases, gelatinases, and elastases); or 4) the pathological angiogenesis as in proliferative retinopathy and tumor metastasis.
  • proteolytic enzymes such as the matrix metalloproteinases (e.g., collagenases, gelatinases, and elastases)
  • the pathological angiogenesis as in proliferative retinopathy and tumor metastasis.
  • Exemplary proliferative diseases include cancers (i.e., “malignant neoplasms”), benign neoplasms, angiogenesis, inflammatory diseases, and autoimmune diseases.
  • angiogenesis refers to the physiological process through which new blood vessels form from pre-existing vessels. Angiogenesis is distinct from vasculogenesis, which is the de novo formation of endothelial cells from mesoderm cell precursors. The first vessels in a developing embryo form through vasculogenesis, after which angiogenesis is responsible for most blood vessel growth during normal or abnormal development. Angiogenesis is a vital process in growth and development, as well as in wound healing and in the formation of granulation tissue.
  • angiogenesis is also a fundamental step in the transition of tumors from a benign state to a malignant one, leading to the use of angiogenesis inhibitors in the treatment of cancer.
  • Angiogenesis may be chemically stimulated by angiogenic proteins, such as growth factors (e.g., VEGF).
  • VEGF growth factors
  • “Pathological angiogenesis” refers to abnormal (e.g., excessive or insufficient) angiogenesis that amounts to and/or is associated with a disease.
  • the terms “neoplasm” and “tumor” are used herein interchangeably and refer to an abnormal mass of tissue wherein the growth of the mass surpasses and is not coordinated with the growth of a normal tissue.
  • a neoplasm or tumor may be “benign” or “malignant,” depending on the following characteristics: degree of cellular differentiation (including morphology and functionality), rate of growth, local invasion, and metastasis.
  • a “benign neoplasm” is generally well differentiated, has characteristically slower growth than a malignant neoplasm, and remains localized to the site of origin.
  • a benign neoplasm does not have the capacity to infiltrate, invade, or metastasize to distant sites.
  • Exemplary benign neoplasms include, but are not limited to, lipoma, chondroma, adenomas, acrochordon, senile angiomas, seborrheic keratoses, lentigos, and sebaceous hyperplasias.
  • certain “benign” tumors may later give rise to malignant neoplasms, which may result from additional genetic changes in a subpopulation of the tumor’s neoplastic cells, and these tumors
  • U1197.70243WO00 26/193 #13587456v2 are referred to as “pre-malignant neoplasms.”
  • An exemplary pre-malignant neoplasm is a teratoma.
  • a “malignant neoplasm” is generally poorly differentiated (anaplasia) and has characteristically rapid growth accompanied by progressive infiltration, invasion, and destruction of the surrounding tissue.
  • a malignant neoplasm generally has the capacity to metastasize to distant sites.
  • metastasis refers to the spread or migration of cancerous cells from a primary or original tumor to another organ or tissue and is typically identifiable by the presence of a “secondary tumor” or “secondary cell mass” of the tissue type of the primary or original tumor and not of that of the organ or tissue in which the secondary (metastatic) tumor is located.
  • a prostate cancer that has migrated to bone is said to be metastasized prostate cancer and includes cancerous prostate cancer cells growing in bone tissue.
  • cancer refers to a class of diseases characterized by the development of abnormal cells that proliferate uncontrollably and have the ability to infiltrate and destroy normal body tissues.
  • Exemplary cancers include, but are not limited to, acoustic neuroma; adenocarcinoma; adrenal gland cancer; anal cancer; angiosarcoma (e.g., lymphangiosarcoma, lymphangioendotheliosarcoma, hemangiosarcoma); appendix cancer; benign monoclonal gammopathy; biliary cancer (e.g., cholangiocarcinoma); bladder cancer; breast cancer (e.g., adenocarcinoma of the breast, papillary carcinoma of the breast, mammary cancer, medullary carcinoma of the breast); brain cancer (e.g., meningioma, glioblastomas, glioma (e.g., astrocytoma, oligodendroglioma), medul
  • angiosarcoma e.g., lymphangiosarcoma, lymphangioendotheliosarcoma, hemangiosar
  • U1197.70243WO00 27/193 #13587456v2 lymphoma splenic marginal zone B-cell lymphoma), primary mediastinal B-cell lymphoma, Burkitt lymphoma, lymphoplasmacytic lymphoma (i.e., Waldenström’s macroglobulinemia), hairy cell leukemia (HCL), immunoblastic large cell lymphoma, precursor B-lymphoblastic lymphoma and primary central nervous system (CNS) lymphoma; and T-cell NHL such as precursor T-lymphoblastic lymphoma/leukemia, peripheral T-cell lymphoma (PTCL) (e.g., cutaneous T-cell lymphoma (CTCL) (e.g., mycosis fungoides, Sezary syndrome), angioimmunoblastic T-cell lymphoma, extranodal natural killer T-cell lymphoma, enteropathy type T-cell lymphoma, subcutaneous panniculitis-like T
  • Wilms tumor, renal cell carcinoma); liver cancer (e.g., hepatocellular cancer (HCC), malignant hepatoma); lung cancer (e.g., bronchogenic carcinoma, small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), adenocarcinoma of the lung); leiomyosarcoma (LMS); mastocytosis (e.g., systemic mastocytosis); muscle cancer; myelodysplastic syndrome (MDS); mesothelioma; myeloproliferative disorder (MPD) (e.g., polycythemia vera (PV), essential thrombocytosis (ET), agnogenic myeloid metaplasia (AMM) a.k.a.
  • HCC hepatocellular cancer
  • lung cancer e.g., bronchogenic carcinoma, small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), adenocarcinoma of the lung
  • myelofibrosis MF
  • chronic idiopathic myelofibrosis chronic myelocytic leukemia (CML), chronic neutrophilic leukemia (CNL), hypereosinophilic syndrome (HES)
  • neuroblastoma e.g., neurofibromatosis (NF) type 1 or type 2, schwannomatosis
  • neuroendocrine cancer e.g., gastroenteropancreatic neuroendoctrine tumor (GEP-NET), carcinoid tumor
  • osteosarcoma e.g.,bone cancer
  • ovarian cancer e.g., cystadenocarcinoma, ovarian embryonal carcinoma, ovarian adenocarcinoma
  • papillary adenocarcinoma pancreatic cancer
  • pancreatic cancer e.g., pancreatic andenocarcinoma, intraductal papillary mucinous neoplasm (IPMN), Islet cell tumors
  • U1197.70243WO00 28/193 #13587456v2 characterized by signs of pain (dolor, from the generation of noxious substances and the stimulation of nerves), heat (calor, from vasodilatation), redness (rubor, from vasodilatation and increased blood flow), swelling (tumor, from excessive inflow or restricted outflow of fluid), and/or loss of function (functio laesa, which can be partial or complete, temporary or permanent.
  • Inflammation takes on many forms and includes, but is not limited to, acute, adhesive, atrophic, catarrhal, chronic, cirrhotic, diffuse, disseminated, exudative, fibrinous, fibrosing, focal, granulomatous, hyperplastic, hypertrophic, interstitial, metastatic, necrotic, obliterative, parenchymatous, plastic, productive, proliferous, pseudomembranous, purulent, sclerosing, seroplastic, serous, simple, specific, subacute, suppurative, toxic, traumatic, and/or ulcerative inflammation.
  • inflammatory disease may also refer to a dysregulated inflammatory reaction that causes an exaggerated response by macrophages, granulocytes, and/or T-lymphocytes leading to abnormal tissue damage and/or cell death.
  • An inflammatory disease can be either an acute or chronic inflammatory condition and can result from infections or non-infectious causes.
  • Inflammatory diseases include, without limitation, atherosclerosis, arteriosclerosis, autoimmune disorders, multiple sclerosis, systemic lupus erythematosus, polymyalgia rheumatica (PMR), gouty arthritis, degenerative arthritis, tendonitis, bursitis, psoriasis, cystic fibrosis, arthrosteitis, rheumatoid arthritis, inflammatory arthritis, Sjogren’s syndrome, giant cell arteritis, progressive systemic sclerosis (scleroderma), ankylosing spondylitis, polymyositis, dermatomyositis, pemphigus, pemphigoid, diabetes (e.g., Type I), myasthenia gravis, Hashimoto’s thyroiditis, Graves’ disease, Goodpasture’s disease, mixed connective tissue disease, sclerosing cholangitis, inflammatory bowel disease, Crohn’s disease, ulcerative colitis, per
  • An ocular inflammatory disease includes, but is not limited to, post-surgical inflammation.
  • Additional exemplary inflammatory conditions include, but are not limited to, inflammation associated with acne, anemia (e.g., aplastic anemia, hemolytic autoimmune anemia), asthma, arteritis (e.g., polyarteritis, temporal arteritis, periarteritis nodosa, Takayasu’s arteritis), arthritis (e.g., crystalline arthritis, osteoarthritis, psoriatic arthritis, gouty arthritis, reactive arthritis, rheumatoid arthritis and Reiter’s arthritis), ankylosing spondylitis, amylosis, amyotrophic lateral sclerosis, autoimmune diseases, allergies or allergic reactions, atherosclerosis, bronchitis, bursitis, chronic prostatitis, conjunctivitis, Chagas disease
  • anemia e.g., aplastic anemia, hemolytic autoimmune anemia
  • asthma e
  • the inflammatory disorder is selected from arthritis (e.g., rheumatoid arthritis), inflammatory bowel disease, inflammatory bowel syndrome, asthma, psoriasis, endometriosis, interstitial cystitis and prostatitis.
  • the inflammatory condition is an acute inflammatory condition (e.g., for example, inflammation resulting from infection).
  • the inflammatory condition is a chronic inflammatory condition (e.g., conditions resulting from asthma, arthritis and inflammatory bowel disease).
  • the compounds may also be useful in treating inflammation associated with trauma and non-inflammatory myalgia.
  • the compounds disclosed herein may also be useful in treating inflammation associated with cancer.
  • autoimmune disease refers to a disease arising from an inappropriate immune response of the body of a subject against substances and tissues normally present in the body. In other words, the immune system mistakes some part of the body as a pathogen and attacks its own cells. This may be restricted to certain organs (e.g., in autoimmune thyroiditis) or involve a particular tissue in different places (e.g., Goodpasture’s disease which may affect the basement membrane in both the lung and kidney).
  • the treatment of autoimmune diseases is typically with immunosuppression, e.g., medications which decrease the immune response.
  • Exemplary autoimmune diseases include, but are not limited to, glomerulonephritis, Goodpasture’s syndrome, necrotizing vasculitis, lymphadenitis, peri-arteritis nodosa, systemic lupus erythematosis, rheumatoid arthritis, psoriatic arthritis, , psoriasis, ulcerative colitis, systemic sclerosis, dermatomyositis/polymyositis, anti-phospholipid antibody syndrome, scleroderma, pemphigus vulgaris, ANCA-associated vasculitis (e.g., Wegener’s granulomatosis, microscopic polyangiitis), uveitis, Sjogren’s syndrome, Crohn’s disease, Reiter’s syndrome, ankylosing spondylitis, Lyme disease, Guillain-Barré syndrome, Hashimoto’s thyroiditis, and cardiomyopathy.
  • liver disease refers to damage to or a disease of the liver.
  • liver disease include intrahepatic cholestasis (e.g., alagille syndrome, biliary liver cirrhosis), fatty liver (e.g., alcoholic fatty liver, Reye’s syndrome), hepatic vein thrombosis, hepatolenticular degeneration (i.e., Wilson’s disease), hepatomegaly, liver abscess (e.g., amebic liver abscess), liver cirrhosis (e.g., alcoholic, biliary, and experimental liver cirrhosis), alcoholic liver diseases (e.g., fatty liver, hepatitis, cirrhosis), parasitic liver disease (e.g., hepatic echinococcosis, fascioliasis, amebic liver abscess), jaundice (e.g.,
  • Immune disorders include, but are not limited to, arthritis (including rheumatoid arthritis, spondyloarthopathies, gouty arthritis, degenerative joint diseases such as osteoarthritis, systemic lupus erythematosus, Sjogren’s syndrome, ankylosing spondylitis, undifferentiated spondylitis, Behcet’s disease, haemolytic autoimmune anaemias, multiple sclerosis, amyotrophic lateral sclerosis, amylosis, acute painful shoulder, psoriatic, and juvenile arthritis), asthma, atherosclerosis, osteoporosis, bronchitis, tendonitis, bursitis, skin condition (e.g., psoriasis, eczema, burns, dermatitis, pruritus (itch)), enuresis, eosinophilic disease, gastrointestinal disorder (e.g., selected from p
  • a “microbial infection” refers to an infection with a microorganism, such as a fungus, bacteria or virus.
  • the microbial infection is an infection with a fungus, i.e., a fungal infection.
  • the microbial infection is an infection with a virus, i.e., a viral infection.
  • the microbial infection is an infection with a bacteria, i.e., a bacterial infection.
  • Various microbial infections include, but are not limited to, skin infections, GI infections, urinary tract infections, genito-urinary infections, sepsis, blood infections, and systemic infections.
  • particle refers to a small object, fragment, or piece of a substance that may be a single element, inorganic material, organic material, or mixture thereof.
  • particles include polymeric particles, single-emulsion particles, double-emulsion particles, coacervates, liposomes, microparticles, nanoparticles, macroscopic particles, pellets, crystals, aggregates, composites, pulverized, milled or otherwise disrupted matrices, and cross-linked protein or polysaccharide particles, each of which have an average characteristic dimension of about less than about 1 mm and at least 1 nm, where the characteristic dimension, or “critical dimension,” of the particle is the smallest cross-sectional dimension of the particle.
  • a particle may be composed of a single substance or multiple substances.
  • the particle is not a viral particle.
  • the particle is not a liposome.
  • the particle is not a micelle.
  • the particle is substantially solid throughout.
  • the particle is a nanoparticle.
  • the particle is a microparticle.
  • the term “nanoparticle” refers to a particle having an average (e.g., mean) dimension (e.g., diameter) of between about 1 nanometer (nm) and about 1 micrometer ( ⁇ m) (e.g., between about 1 nm and about 300 nm, between about 1 nm and about 100 nm, between about 1 nm and about 30 nm, between about 1 nm and about 10 nm, or between about 1 nm and about 3 nm), inclusive.
  • microparticle refers to a particle having an average (e.g., mean) dimension (e.g., diameter) of between about 1 micrometer ( ⁇ m) and about 1 millimeter (mm) (e.g., between about 1 ⁇ m and about 100 ⁇ m, between about 1 ⁇ m and about 30 ⁇ m, between about 1 ⁇ m and about 10 ⁇ m, or between about 1 ⁇ m and about 3 ⁇ m), inclusive.
  • the “hydrodynamic diameter” of a particle refers to the diameter of a solid sphere that would exhibit the same hydrodynamic friction as the particle (e.g., the diameter of a solid sphere that diffuses at the same rate as the particle).
  • Hydrodynamic diameter can be measured through various techniques including dynamic light scattering (DLS) and nanoparticle tracking analysis (NTA).
  • polydispersity index or “PDI” refer to the degree of non-uniformity of a size distribution of particles (e.g., the broadness of a molecular weight distribution). PDI can be measured through various techniques including dynamic light scattering (DLS).
  • zeta potential refers to the potential difference between the surface of a particle and the surrounding liquid the particles are dispersed in (e.g., the surface charge of nanoparticles in solution). Zeta potential can be measured through various techniques including electrophoretic light scattering and electroacoustic phenomenon.
  • Ka refers to the negative decadic logarithm of the ionization constant (Ka) of an acid; equal to the pH value at which equal concentrations of the acid and conjugate base forms of a substance (often a buffer) are present.
  • Ka ionization constant
  • dendrimer refers to a molecular architecture with an interior core, interior layers (or “generations”) of repeating units which are attached to and extend from this interior core, each layer having one or more branching points, and the outermost generation having terminal functional groups on an exterior surface.
  • dendrimers include, but are not limited to, poly(amidoamine) (PAMAM), polyester, polylysine, and poly(propylene imine) (PPI).
  • terminal functional groups of a dendrimer include one or more hydroxyl groups, one or more amine groups, and/or one or more carboxyl groups.
  • PAMAM dendrimer means poly(amidoamine) dendrimer, which may contain different cores, with amidoamine building blocks. The method for making them is known to those of
  • This PAMAM core- shell architecture grows linearly in diameter as a function of added shells (generations). Meanwhile, the surface groups amplify exponentially at each generation according to dendritic-branching mathematics.
  • PAMAM dendrimers can have carboxylic acid, amine and/or hydroxyl terminations and can be any generation of dendrimers including, but not limited to, generation 1 PAMAM dendrimers, generation 2 PAMAM dendrimers, generation 3 PAMAM dendrimers, generation 4 PAMAM dendrimers, generation 5 PAMAM dendrimers, generation 6 PAMAM dendrimers, generation 7 PAMAM dendrimers, generation 8 PAMAM dendrimers, generation 9 PAMAM dendrimers, or generation 10 PAMAM dendrimers.
  • the number of terminal sites on a dendrimer can depend on the particular dendrimeric scaffold and its generation.
  • a dendrimer is based on a generation 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 PAMAM dendrimeric scaffold, which have 4, 8, 16, 32, 64, 128, 256, 512, 1024, 2048, and 4096 terminal sites, respectively.
  • DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS [0136] The aspects described herein are not limited to specific embodiments, systems, compositions, methods, or configurations, and as such can, of course, vary. The terminology used herein is for the purpose of describing particular aspects only and, unless specifically defined herein, is not intended to be limiting.
  • PAMAM n
  • U1197.70243WO00 35/193 #13587456v2 p is an integer between 1 and 2 (n+2) , inclusive; q is an integer between 0 and 2 (n+2) -1, inclusive; and the sum of p and q is 2 (n+2) .
  • PAMAM nth generation polyamidoamine
  • U1197.70243WO00 37/193 #13587456v2 q is an integer between 0 and 2 (n+2) -1, inclusive; and the sum of p and q is 2 (n+2) .
  • the present disclosure provides a compound of Formula (III): or a pharmaceutically acceptable salt thereof, wherein: Z n is an nth generation polyamidoamine (PAMAM) dendrimer; n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10;
  • PAMAM polyamidoamine
  • PAMAM
  • PAMAM
  • the present disclosure provides a compound of Formula (VI): or a pharmaceutically acceptable salt thereof, wherein: Z n is an nth generation polyamidoamine (PAMAM) dendrimer; n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10;
  • PAMAM polyamidoamine
  • PAMAM n
  • U1197.70243WO00 41/193 #13587456v2 optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene; each instance of R 1 is independently hydrogen or optionally substituted alkyl; p is an integer between 1 and 2 (n+2) , inclusive; q is an integer between 0 and 2 (n+2) -1, inclusive; and the sum of p and q is 2 (n+2) .
  • PAMAM polya
  • Z n is an nth generation polyamidoamine (PAMAM) dendrimer.
  • PAMAM polyamidoamine
  • the nth generation PAMAM dendrimer is prepared by a two-step iterative reaction sequence that produces concentric shells (generations) of dendritic 0-alanine units around a central initiator core.
  • the nth generation PAMAM dendrimer is a generation 1 PAMAM dendrimer, generation 2 PAMAM dendrimer, generation 3 PAMAM dendrimer, generation 4 PAMAM dendrimer, generation 5 PAMAM dendrimer, generation 6 PAMAM dendrimer, generation 7 PAMAM dendrimer, generation 8 PAMAM dendrimer, generation 9 PAMAM dendrimer, or generation 10 PAMAM dendrimer.
  • the nth generation PAMAM dendrimer is a generation 6 PAMAM dendrimer.
  • Z n is an nth generation PAMAM dendrimer (i.e., Z n is the core of an nth generation PAMAM dendrimer, and does not include the terminal groups of the dendrimer).
  • Z 0 is a 0th generation dendrimer (i.e., Z 0 is the core of a 0th generation PAMAM dendrimer).
  • Z 0 is a 0th generation dendrimer (i.e., Z 0 is the core of a 0th generation PAMAM dendrimer) of formula:
  • Z 1 is a 1st generation PAMAM dendrimer (i.e., Z 1 is the core of a 1st generation PAMAM dendrimer).
  • Z 1 is a 1st generation PAMAM dendrimer (i.e., Z 1 is the core of a 1st generation PAMAM dendrimer) of formula: .
  • the compound of Formula (I) or Formula (I′), wherein: Z 1 is an 1st generation polyamidoamine (PAMAM) dendrimer; n is 1; each instance of Y is independently can be represented by the formula: ,
  • n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.
  • n is 0, 1, 2, 3, 4, 5, 6, 7, 8, or 9.
  • n is 0, 1, 2, 3, 4, 5, 6, 7, or 8.
  • n is 0, 1, 2, 3, 4, 5, 6, or 7.
  • n is 0, 1, 2, 3, 4, 5, or 6.
  • n is 0, 1, 2, 3, 4, or 5.
  • n is 0, 1, 2, 3, or 4.
  • n is 0, 1, 2, or 3.
  • n is 0, 1, or 2. In some embodiments, n is 0 or 1.
  • n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, n is 1, 2, 3, 4, 5, 6, 7, 8, or 9. In some embodiments, n is 1, 2, 3, 4, 5, 6, 7, or 8. In some embodiments, n is 1, 2, 3, 4, 5, 6, or 7. In some embodiments, n is 1, 2, 3, 4, 5, or 6. In some embodiments, n is 1, 2, 3, 4, or 5. In some embodiments, n is 1, 2, 3, or 4. In some embodiments, n is 1, 2 or 3. In some embodiments, n is 1 or 2. In some embodiments, n is 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, n is 2, 3, 4, 5, 6, 7, 8, or 9. In some embodiments, n is 2, 3, 4, 5, 6, 7, or 8.
  • n is 2, 3, 4, 5, 6, or 7. In some embodiments, n is 2, 3, 4, 5, or 6. In some embodiments, n is 2, 3, 4, or 5. In some embodiments, n is 2, 3, or 4. In some embodiments, n is 2 or 3. In some embodiments, n is 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, n is 3, 4, 5, 6, 7, 8, or 9. In some embodiments, n is 3, 4, 5, 6, 7, or 8. In some embodiments, n is 3, 4, 5, 6, or 7. In some embodiments, n is 3, 4, 5, or 6. In some embodiments, n is 3, 4, or 5. In some embodiments, n is 3 or 4. [0157] In some embodiments, n is 4 or greater.
  • n is 4, 5, 6, 7, 8, 9, or 10. In some embodiments, n is 4, 5, 6, 7, 8, or 9. In some embodiments, n is 4, 5, 6, 7, or 8. In some embodiments, n is 4, 5, 6, or 7. In some embodiments, n is 4, 5, or 6. In some embodiments, n is 4 or 5. In some embodiments, n is 5 or greater. In some embodiments, n is 5, 6, 7, 8, 9, or 10. In some embodiments, n is 5, 6, 7, 8, or 9. In some embodiments, n is 5, 6, 7, or 8. In some embodiments, n is 5, 6, or 7. In some embodiments, n is 5 or 6. In some embodiments, n is 6 or greater. In some embodiments, n is 6, 7, 8, 9, or 10.
  • n is 6, 7, 8, or 9. In some embodiments, n is 6, 7, or 8. In some embodiments, n is 6 or 7. In some embodiments, n is 7 or greater. In some embodiments, n is 7, 8, 9, or 10. In some embodiments, n is 7, 8, or 9. In some embodiments, n is 7 or 8. In some embodiments, n is 8 or greater. In some embodiments, n is 8, 9, or 10. In some embodiments, n is 8 or 9. In some embodiments, n is 9 or greater. In some embodiments, n is 9 or 10. [0158] In some embodiments, n is 0. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4.
  • n is 5. In some embodiments, n is 6. In some embodiments, n is 7. In some embodiments, n is 8. In some embodiments, n is 9. In some embodiments, n is 10. [0159] In some embodiments, n is 0, such that Z n is Z 0 , and Z 0 is a 0th generation PAMAM dendrimer. In some embodiments, n is 1, such that Z n is Z 1 , and Z 1 is a 1st generation PAMAM dendrimer. In some embodiments, n is 2, such that Z n is Z 2 , and Z 2 is a 2nd generation PAMAM dendrimer.
  • n is 3, such that Z n is Z 3 , and Z 3 is a 3rd generation PAMAM dendrimer. In some embodiments, n is 4, such that Z n is Z 4 , and Z 4 is a 4th generation PAMAM
  • n is 5, such that Z n is Z 5 , and Z 5 is a 5th generation PAMAM dendrimer.
  • n is 6, such that Z n is Z 6 , and Z 6 is a 6th generation PAMAM dendrimer.
  • n is 7, such that Z n is Z 7 , and Z 7 is a 7th generation PAMAM dendrimer.
  • n is 8, such that Z n is Z 8 , and Z 8 is a 8th generation PAMAM dendrimer.
  • n is 9, such that Z n is Z 9 , and Z 9 is a 9th generation PAMAM dendrimer. In some embodiments, n is 10, such that Z n is Z 10 , and Z 10 is a 10th generation PAMAM dendrimer. [0160] In some embodiments, Z n is Z 4 , Z 5 , Z 6 , Z 7 , Z 8 , Z 9 , or Z 10 . In some embodiments, Z n is Z 4 , Z 5 , Z 6 , Z 7 , Z 8 , or Z 9 . In some embodiments, Z n is Z 4 , Z 5 , Z 6 , Z 7 , or Z 8 .
  • Z n is Z 4 , Z 5 , Z 6 , or Z 7 . In some embodiments, Z n is Z 4 , Z 5 , or Z 6 . In some embodiments, Z n is Z 5 , Z 6 , Z 7 , Z 8 , Z 9 , or Z 10 . In some embodiments, Z n is Z 5 , Z 6 , Z 7 , Z 8 , or Z 9 . In some embodiments, Z n is Z 5 , Z 6 , Z 7 , or Z 8 . In some embodiments, Z n is Z 5 , Z 6 , or Z 7 . In some embodiments, Z n is Z 5 or Z 6 .
  • Z n is Z 6 , Z 7 , Z 8 , Z 9 , or Z 10 . In some embodiments, Z n is Z 6 , Z 7 , Z 8 , or Z 9 . In some embodiments, Z n is Z 6 , Z 7 , or Z 8 . In some embodiments, Z n is Z 6 or Z 7 . [0161] As generally described herein, p is an integer between 1 and 2 (n+2) , inclusive; q is an integer between 0 and 2 (n+2) -1, inclusive; and the sum of p and q is 2 (n+2) . [0162] In some embodiments, n is 0, such that the sum of p and q is 4.
  • n is 1, such that the sum of p and q is 8. In some embodiments, n is 2, such that the sum of p and q is 16. In some embodiments, n is 3, such that the sum of p and q is 32. [0163] In some embodiments, n is 4, such that the sum of p and q is 64. In some embodiments, the sum of p and q is 64. In some embodiments, the sum of p and q is 64, p is an integer between 1 and 10, inclusive, and q is an integer between 54 and 63, inclusive. In some embodiments, the sum of p and q is 64, p is an integer between 1 and 5, inclusive, and q is an integer between 59 and 63, inclusive.
  • n is 5, such that the sum of p and q is 128. In some embodiments, the sum of p and q is 128. In some embodiments, the sum of p and q is 128, p is an integer between 1 and 20, inclusive, and q is an integer between 108 and 127, inclusive. In some embodiments, the sum of p and q is 128, p is an integer between 1 and 15, inclusive, and q is an integer between 113 and 127, inclusive. In some embodiments, the sum of p and q is 128, p is an integer between 1 and 10, inclusive, and q is an integer between 118 and 127, inclusive.
  • the sum of p and q is 128, p is an integer between 1 and 5, inclusive, and q is an integer between 123 and 127, inclusive. [0165] In some embodiments, n is 6, such that the sum of p and q is 256. In some embodiments, the sum of p and q is 256. In some embodiments, the sum of p and q is 256, p is an integer between 1 and 40, inclusive, and q is an integer between 216 and 255, inclusive. In some embodiments, the sum of p and q is 256, p is an integer between 5 and 35, inclusive, and q is an integer between 221 and 251, inclusive. In some embodiments, the sum of p and q is 256, p is an integer between 8 and 32,
  • q is an integer between 224 and 248, inclusive.
  • the sum of p and q is 256, p is an integer between 10 and 30, inclusive, and q is an integer between 226 and 246, inclusive.
  • the sum of p and q is 256, p is an integer between 12 and 28, inclusive, and q is an integer between 228 and 244, inclusive.
  • the sum of p and q is 256, p is an integer between 14 and 26, inclusive, and q is an integer between 230 and 242, inclusive.
  • the sum of p and q is 256, p is an integer between 15 and 25, inclusive, and q is an integer between 231 and 241, inclusive. In some embodiments, the sum of p and q is 256, p is an integer between 16 and 24, inclusive, and q is an integer between 232 and 240, inclusive. In some embodiments, the sum of p and q is 256, p is an integer between 17 and 23, inclusive, and q is an integer between 233 and 239, inclusive. In some embodiments, the sum of p and q is 256, p is an integer between 18 and 22, inclusive, and q is an integer between 234 and 238, inclusive.
  • the sum of p and q is 256, p is an integer between 19 and 21, inclusive, and q is an integer between 235 and 237, inclusive. [0166] In some embodiments, the sum of p and q is 256, p is an integer between 1 and 30, inclusive, and q is an integer between 226 and 255, inclusive. In some embodiments, the sum of p and q is 256, p is an integer between 3 and 27, inclusive, and q is an integer between 229 and 253, inclusive. In some embodiments, the sum of p and q is 256, p is an integer between 5 and 25, inclusive, and q is an integer between 231 and 251, inclusive.
  • the sum of p and q is 256, p is an integer between 7 and 23, inclusive, and q is an integer between 233 and 249, inclusive. In some embodiments, the sum of p and q is 256, p is an integer between 9 and 21, inclusive, and q is an integer between 235 and 247, inclusive. In some embodiments, the sum of p and q is 256, p is an integer between 10 and 20, inclusive, and q is an integer between 236 and 246, inclusive. In some embodiments, the sum of p and q is 256, p is an integer between 11 and 19, inclusive, and q is an integer between 237 and 245, inclusive.
  • the sum of p and q is 256, p is an integer between 12 and 18, inclusive, and q is an integer between 238 and 244, inclusive. In some embodiments, the sum of p and q is 256, p is an integer between 13 and 17, inclusive, and q is an integer between 239 and 243, inclusive. In some embodiments, the sum of p and q is 256, p is an integer between 14 and 16, inclusive, and q is an integer between 240 and 242, inclusive. [0167] In some embodiments, p is 1 and q is 255. In some embodiments, p is 2 and q is 254. In some embodiments, p is 3 and q is 253. In some embodiments, p is 4 and q is 252.
  • p is 5 and q is 251. In some embodiments, p is 6 and q is 250. In some embodiments, p is 7 and q is 249. In some embodiments, p is 8 and q is 248. In some embodiments, p is 9 and q is 247. In some embodiments, p is 10 and q is 246. In some embodiments, p is 11 and q is 245. In some embodiments, p is 12 and q is 244. In some embodiments, p is 13 and q is 243. In some embodiments, p is 14 and q is 242. In some embodiments, p is 15 and q is 241. In some embodiments, p is 16 and q is 240.
  • p is 17 and q is 239. In some embodiments, p is 18 and q is 238. In some embodiments, p is 19 and q is 237. In some embodiments, p is 20 and q is 236. In some embodiments,
  • U1197.70243WO00 49/193 #13587456v2 p is 21 and q is 235. In some embodiments, p is 22 and q is 234. In some embodiments, p is 23 and q is 233. In some embodiments, p is 24 and q is 232. In some embodiments, p is 25 and q is 231. In some embodiments, p is 26 and q is 230. In some embodiments, p is 27 and q is 229. In some embodiments, p is 28 and q is 228. In some embodiments, p is 29 and q is 227. In some embodiments, p is 30 and q is 226. In some embodiments, p is 31 and q is 225.
  • p is 32 and q is 224. In some embodiments, p is 33 and q is 223. In some embodiments, p is 34 and q is 222. In some embodiments, p is 35 and q is 221. In some embodiments, p is 36 and q is 220. In some embodiments, p is 37 and q is 219. In some embodiments, p is 38 and q is 218. In some embodiments, p is 39 and q is 217. In some embodiments, p is 40 and q is 216. In some embodiments, p is 15 and q is 241; or p is 20 and q is 236.
  • n is 7, such that the sum of p and q is 512. In some embodiments, the sum of p and q is 512. In some embodiments, the sum of p and q is 512, p is an integer between 1 and 80, inclusive, and q is an integer between 432 and 511, inclusive. In some embodiments, the sum of p and q is 512, p is an integer between 1 and 40, inclusive, and q is an integer between 472 and 511, inclusive. [0169] In some embodiments, n is 8, such that the sum of p and q is 1024. In some embodiments, the sum of p and q is 1024.
  • the sum of p and q is 1024, p is an integer between 1 and 160, inclusive, and q is an integer between 864 and 1023, inclusive. In some embodiments, the sum of p and q is 512, p is an integer between 1 and 80, inclusive, and q is an integer between 944 and 1023, inclusive. [0170] In some embodiments, n is 9, such that the sum of p and q is 2048. In some embodiments, the sum of p and q is 2048. In some embodiments, the sum of p and q is 2048, p is an integer between 1 and 320, inclusive, and q is an integer between 1728 and 2047, inclusive.
  • the sum of p and q is 2048, p is an integer between 1 and 160, inclusive, and q is an integer between 1888 and 2047, inclusive.
  • n is 10, such that the sum of p and q is 4096.
  • the sum of p and q is 4096.
  • the sum of p and q is 4096, p is an integer between 1 and 640, inclusive, and q is an integer between 3456 and 4095, inclusive.
  • the sum of p and q is 4096, p is an integer between 1 and 320, inclusive, and q is an integer between 3776 and 4095, inclusive.
  • p is at least 1. In some embodiments, p is at least 2. In some embodiments, p is at least 3. In some embodiments, p is at least 4. In some embodiments, p is at least 5. In some embodiments, p is at least 6. In some embodiments, p is at least 7. In some embodiments, p is at least 8. In some embodiments, p is at least 9. In some embodiments, p is at least 10. In some embodiments, p is at least 11. In some embodiments, p is at least 12. In some embodiments, p is at least 13. In some embodiments, p is at least 14. In some embodiments, p is at least 15. In some
  • p is at least 16. In some embodiments, p is at least 17. In some embodiments, p is at least 18. In some embodiments, p is at least 19. In some embodiments, p is at least 20.
  • at least one instance of L is optionally substituted C 1-20 alkylene, optionally substituted 5-6 membered heteroarylene, or a combination thereof, optionally wherein one or more backbone
  • At least one instance of L is optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1, 2, 3, 4, or 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –O–.
  • at least one instance of L is optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1 backbone carbon atom in the optionally substituted linear C 1-20 alkylene is replaced with –O–.
  • At least one instance of L is optionally substituted linear C 1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 2 backbone carbon atoms in the optionally substituted linear C 1-20 alkylene are replaced with –O–.
  • at least one instance of L is optionally substituted linear C 1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 3 backbone carbon atoms in the optionally substituted linear C 1-20 alkylene are replaced with –O–.
  • at least one instance of L is optionally substituted linear C 1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof,
  • U1197.70243WO00 52/193 #13587456v2 wherein at least 4 backbone carbon atoms in the optionally substituted linear C 1-20 alkylene are replaced with –O–.
  • at least one instance of L is optionally substituted linear C 1- 20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 5 backbone carbon atoms in the optionally substituted linear C 1- 20 alkylene are replaced with –O–.
  • At least one instance of L is optionally substituted linear C 1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1, 2, 3, 4, or 5 backbone carbon atoms in the optionally substituted linear C1- 20 alkylene are replaced with –NR 1 –.
  • at least one instance of L is optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1 backbone carbon atoms in the optionally substituted linear C1-20 alkylene is replaced with –NR 1 –.
  • At least one instance of L is optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 2 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –NR 1 –.
  • at least one instance of L is optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 3 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –NR 1 –.
  • At least one instance of L is optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 4 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –NR 1 –.
  • at least one instance of L is optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 5 backbone carbon atoms in the optionally substituted linear C1- 20 alkylene are replaced with –NR 1 –.
  • at least one instance of L is optionally substituted linear C 1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 3
  • at least one instance of L is optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1, 2, 3, 4, or 5 backbone carbon atoms in the optionally substituted linear C1- 20 alkylene are replaced with optionally substituted heteroarylene.
  • At least one instance of L is optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1 backbone carbon atom in the optionally substituted linear C1-20 alkylene is replaced with optionally substituted heteroarylene.
  • at least one instance of L comprises optionally substituted C 1-20 alkylene, optionally substituted 5-6 membered heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in
  • At least one instance of L comprises optionally substituted linear C 1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1, 2, 3, 4, or 5 backbone carbon atoms in the optionally substituted linear C 1-20 alkylene are replaced with –O–.
  • at least one instance of L comprises optionally substituted linear C 1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1 backbone carbon atom in the optionally substituted linear C 1-20 alkylene is replaced with –O–.
  • At least one instance of L comprises optionally substituted linear C 1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 2 backbone carbon atoms in the optionally substituted linear C 1-20 alkylene are replaced with –O–. In some embodiments, at least one instance of L comprises optionally
  • at least one instance of L comprises optionally substituted linear C 1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 4 backbone carbon atoms in the optionally substituted linear C 1-20 alkylene are replaced with –O–.
  • At least one instance of L comprises optionally substituted linear C 1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –O–.
  • at least one instance of L comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1, 2, 3, 4, or 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –NR 1 –.
  • At least one instance of L comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1 backbone carbon atoms in the optionally substituted linear C1-20 alkylene is replaced with –NR 1 –.
  • at least one instance of L comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 2 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –NR 1 –.
  • At least one instance of L comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 3 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –NR 1 –.
  • at least one instance of L comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 4 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –NR 1 –.
  • At least one instance of L comprises optionally substituted linear C 1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 5 backbone carbon atoms in the optionally substituted linear C 1-20 alkylene are replaced with –NR 1 –.
  • at least one instance of L comprises optionally substituted linear C 1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1
  • at least one instance of L comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1, 2, 3, 4, or 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with optionally substituted heteroarylene.
  • At least one instance of L comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1 backbone carbon atom in the optionally substituted linear C1-20 alkylene is replaced with optionally substituted heteroarylene.
  • at least one instance of L comprises optionally substituted C 1-20 alkylene, optionally wherein one or more backbone carbon atoms in the
  • At least one instance of L comprises optionally substituted linear C1-20 alkylene, wherein at least 1, 2, 3, 4, or 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –O–. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, wherein at least 1 backbone carbon atom in the optionally substituted linear C1-20 alkylene is replaced with –O–. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, wherein at least 2 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –O–.
  • At least one instance of L comprises optionally substituted linear C1-20 alkylene, wherein at least 3 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with – O–. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, wherein at least 4 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –O–. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, wherein at least 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –O–.
  • At least one instance of L comprises optionally substituted linear C 1-20 alkylene, wherein at least 1, 2, 3, 4, or 5 backbone carbon atoms in the optionally substituted linear C 1-20 alkylene are replaced with –NR 1 –. In some embodiments, at least one instance of L comprises optionally substituted linear C 1-20 alkylene, wherein at least 1 backbone carbon atoms in the optionally substituted linear C 1-20 alkylene is replaced with –NR 1 –. In some embodiments, at least one instance of L comprises optionally substituted linear C 1-20 alkylene, wherein at least 2 backbone carbon atoms in the optionally substituted linear C 1-20 alkylene are replaced with –NR 1 –.
  • At least one instance of L comprises optionally substituted linear C 1-20 alkylene, wherein at least 3 backbone carbon atoms in the optionally substituted linear C 1-20 alkylene are replaced with –NR 1 –. In some embodiments, at least one instance of L comprises optionally substituted linear C 1-20 alkylene,
  • At least one instance of L comprises optionally substituted linear C 1-20 alkylene, wherein at least 5 backbone carbon atoms in the optionally substituted linear C 1-20 alkylene are replaced with –NR 1 –.
  • At least one instance of L comprises optionally substituted linear C1-20 alkylene, wherein at least 1 backbone carbon atom in the optionally substituted linear C1-20 alkylene is replaced with optionally substituted heteroarylene. [0195] In some embodiments, at least one instance of L comprises ,
  • each x is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; and each y is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.
  • at least one instance of L comprises 10; and each y is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, at least one instance of L . In some embodiments, at least one instance of L
  • at least one instance of L comprises optionally substituted heteroarylene.
  • at least one instance of L comprises optionally substituted 5-6 membered heteroarylene.
  • at least one instance of L comprises optionally substituted 5 membered heteroarylene.
  • at least one instance of L comprises optionally substituted 6 membered heteroarylene.
  • At least one instance of L comprises optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring heteroatoms selected from O, N, and S. In some embodiments, at least one instance of L comprises optionally substituted 5 membered heteroarylene having 1, 2, or 3 ring heteroatoms selected from O, N, and S. In some embodiments, at least one instance of L comprises optionally substituted 6 membered heteroarylene having 1, 2, or 3 ring heteroatoms selected from O, N, and S. [0198] In some embodiments, at least one instance of L comprises optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms.
  • At least one instance of L comprises optionally substituted 5-6 membered heteroarylene having 1 ring N atom. In some embodiments, at least one instance of L comprises optionally substituted 5-6 membered heteroarylene having 2 ring N atoms. In some embodiments, at least one instance of L comprises optionally substituted 5-6 membered heteroarylene having 3 ring N atoms. In some embodiments, at least one instance of L comprises optionally substituted 5 membered heteroarylene having 1, 2, or 3 ring N atoms. In some embodiments, at least one instance of L comprises optionally substituted 5 membered heteroarylene having 1 ring N atom.
  • At least one instance of L comprises optionally substituted 5 membered heteroarylene having 2 ring N atoms. In some embodiments, at least one instance of L comprises optionally substituted 5 membered heteroarylene having 3 ring N atoms. In some embodiments, at least one instance of L comprises optionally substituted 6 membered heteroarylene having 1, 2, or 3 ring N atoms. In some embodiments, at least one instance of L comprises optionally substituted 6 membered heteroarylene having 1 ring N atom. In some embodiments, at least one instance of L comprises optionally substituted 6 membered heteroarylene having 2 ring N atoms. In some embodiments, at least one instance of L comprises optionally substituted 6 membered heteroarylene having 3 ring N atoms.
  • At least one instance of L comprises , , . In some embodiments, at least one instance of L comprises . In some embodiments, at least one instance of L comprises . In some embodiments, at least one instance of L is , [0201] In some embodiments, at least one instance of L comprises
  • each instance of L independently comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1, 2, 3, 4, or 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –O–.
  • each instance of L independently comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1 backbone carbon atom in the optionally substituted linear C1-20 alkylene is replaced with –O–.
  • each instance of L independently comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 2 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –O–.
  • each instance of L independently comprises optionally substituted linear C 1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 3 backbone carbon atoms in the optionally substituted linear C 1-20 alkylene are replaced with –O–.
  • each instance of L independently comprises optionally substituted linear C 1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 4 backbone carbon atoms in the optionally substituted linear C 1- 20 alkylene are replaced with –O–.
  • each instance of L independently comprises optionally substituted linear C 1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 5 backbone carbon atoms in the optionally substituted linear C 1-20 alkylene are replaced with –O–.
  • each instance of L independently comprises optionally substituted linear C 1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1, 2, 3, 4, or 5 backbone carbon atoms in the optionally substituted linear C 1-20 alkylene are replaced with –NR 1 –.
  • each instance of L independently comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1 backbone carbon atoms in the optionally substituted linear C 1-20 alkylene is replaced with –NR 1 –.
  • each instance of L independently comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 2 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –NR 1 –.
  • each instance of L independently comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 3 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –NR 1 –.
  • each instance of L independently comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 4 backbone carbon atoms in the optionally substituted linear C1- 20 alkylene are replaced with –NR 1 –.
  • each instance of L independently comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –NR 1 –.
  • each instance of L independently comprises optionally substituted linear C 1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a
  • each instance of L independently comprises optionally substituted linear C 1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1, 2, 3, 4, or 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with optionally substituted heteroarylene.
  • each instance of L independently comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1 backbone carbon atom in the optionally substituted linear C1-20 alkylene is replaced with optionally substituted heteroarylene.
  • each instance of L independently comprises optionally substituted linear C1-20 alkylene, wherein at least 1, 2, 3, 4, or 5 backbone carbon atoms in the optionally substituted linear C 1-20 alkylene are replaced with –O–.
  • each instance of L independently comprises optionally substituted linear C 1-20 alkylene, wherein at least 1 backbone carbon atom in the optionally substituted linear C1-20 alkylene is replaced with –O–. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, wherein at least 2 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –O–. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, wherein at least 3 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –O–.
  • each instance of L independently comprises optionally substituted linear C1-20 alkylene, wherein at least 4 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –O–. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, wherein at least 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –O–. [0213] In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, wherein at least 1, 2, 3, 4, or 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –NR 1 –.
  • each instance of L independently comprises optionally substituted linear C1-20 alkylene, wherein at least 1 backbone carbon atoms in the optionally substituted linear C1-20 alkylene is replaced with –NR 1 –. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, wherein at least 2 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –NR 1 –. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, wherein at least 3 backbone carbon atoms in the optionally substituted linear C 1-20 alkylene are replaced with –NR 1 –.
  • each instance of L independently comprises optionally substituted linear C1-20 alkylene, wherein at least 1 backbone carbon atom in the optionally substituted linear C1-20 alkylene is replaced with optionally substituted heteroarylene.
  • each instance of L independently comprises , , 10; and each y is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.
  • each instance of L independently comprises
  • each instance of L independently comprises optionally substituted heteroarylene, optionally wherein one or more backbone carbon atoms in the optionally substituted heteroarylene are independently replaced with –
  • each instance of L independently comprises optionally substituted heteroarylene. In some embodiments, each instance of L independently comprises optionally substituted 5-6 membered heteroarylene. In some embodiments, each instance of L independently comprises optionally substituted 5 membered heteroarylene. In some embodiments, each instance of L independently comprises optionally substituted 6 membered heteroarylene. In some embodiments, each instance of L independently comprises optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring heteroatoms selected from O, N, and S.
  • each instance of L independently comprises optionally substituted 5 membered heteroarylene having 1, 2, or 3 ring heteroatoms selected from O, N, and S. In some embodiments, each instance of L independently comprises optionally substituted 6 membered heteroarylene having 1, 2, or 3 ring heteroatoms selected from O, N, and S. [0219] In some embodiments, each instance of L independently comprises optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms. In some embodiments, each instance of L independently comprises optionally substituted 5-6 membered heteroarylene having 1 ring N atom. In some embodiments, each instance of L independently comprises optionally substituted 5-6 membered heteroarylene having 2 ring N atoms.
  • each instance of L independently comprises optionally substituted 5-6 membered heteroarylene having 3 ring N atoms. In some embodiments, each instance of L independently comprises optionally substituted 5 membered heteroarylene having 1, 2, or 3 ring N atoms. In some embodiments, each instance of L independently comprises optionally substituted 5 membered heteroarylene having 1 ring N atom. In some embodiments, each instance of L independently comprises optionally substituted 5 membered heteroarylene having 2 ring N atoms. In some embodiments, each instance of L independently comprises optionally substituted 5 membered heteroarylene having 3 ring N atoms. In some embodiments, each instance of L independently comprises optionally substituted 6 membered heteroarylene having 1, 2, or 3 ring N atoms.
  • each instance of L independently comprises optionally substituted 6 membered heteroarylene having 1 ring N atom. In some embodiments, each instance of L independently comprises optionally substituted 6 membered heteroarylene having 2 ring N atoms. In some embodiments, each instance of L independently comprises optionally substituted 6 membered heteroarylene having 3 ring N atoms. [0220] In some embodiments, each instance of L independently comprises , , , . In some embodiments, each instance of L independently
  • each instance of L independently comprises . In some embodiments, each instance of L independently comprises . [0221] In some embodiments, each instance of L independently comprises [0222] As generally described herein, each instance of R 1 is independently hydrogen or optionally substituted alkyl. [0223] In some embodiments, at least one instance of R 1 is hydrogen. [0224] In some embodiments, at least one instance of R 1 is optionally substituted alkyl. In some embodiments, at least one instance of R 1 is optionally substituted C1-12 alkyl. In some embodiments, at least one instance of R 1 is optionally substituted C1-6 alkyl.
  • At least one instance of R 1 is optionally substituted C1-3 alkyl. In some embodiments, at least one instance of R 1 is unsubstituted alkyl. In some embodiments, at least one instance of R 1 is unsubstituted C1-12 alkyl. In some embodiments, at least one instance of R 1 is unsubstituted C1-6 alkyl. In some embodiments, at least one instance of R 1 is unsubstituted C1-3 alkyl. In some embodiments, at least one instance of R 1 is
  • each instance of R 1 is independently hydrogen.
  • each instance of R 1 is independently optionally substituted alkyl. In some embodiments, each instance of R 1 is independently optionally substituted C 1-12 alkyl.
  • each instance of R 1 is independently optionally substituted C 1-6 alkyl. In some embodiments, each instance of R 1 is independently optionally substituted C1-3 alkyl. In some embodiments, each instance of R 1 is independently unsubstituted alkyl. In some embodiments, each instance of R 1 is independently unsubstituted C1-12 alkyl. In some embodiments, each instance of R 1 is independently unsubstituted C1-6 alkyl. In some embodiments, each instance of R 1 is independently unsubstituted C1-3 alkyl. In some embodiments, each instance of R 1 is independently unsubstituted linear alkyl. In some embodiments, each instance of R 1 is independently unsubstituted linear C1-12 alkyl.
  • each instance of R 1 is independently unsubstituted linear C1-6 alkyl. In some embodiments, each instance of R 1 is independently unsubstituted linear C1-3 alkyl.
  • X is independently a conjugated agent. In some embodiments, the conjugated agent is a radical of an agent. In some embodiments, each instance of X is independently a radical of an agent. [0228] In some embodiments, the agent is a small molecule compound (e.g., a small molecule organic compound). In some embodiments, the conjugated agent is a radical of a small molecule compound (e.g., a small molecule organic compound).
  • each instance of X is independently a radical of a small molecule compound (e.g., a small molecule organic compound).
  • an agent is a small molecule compound having a molecular weight of less than 2,000 daltons (Da), less than 1,500 Da, less than 1,000 Da, or less than 500 Da. In some embodiments, an agent is a small-molecule compound having a molecular weight of between about 100 and about 2,000 Da.
  • the small-molecule compound has a molecular weight of between about 100 and about 1,500 Da, between about 100 and about 1,000 Da, between about 100 and about 750 Da, between about 100 and about 500 Da, between about 500 and about 2,000 Da, between about 500 and about 1,500 Da, or between about 500 and about 1,000 Da.
  • the conjugated agent is a radical of an agent selected from:
  • the conjugated agent is a radical of LLL12-Hydrazone (18): Hydrazone, 18),
  • LLL12 refers to 5-hydroxy-9,10-dioxo-9,10-dihydroanthracene-1-sulfonamide (CAS No. 1260247-42-4), or a pharmaceutically acceptable salt thereof, which is a signal transducer and activator of transcription 3 (STAT3) inhibitor.
  • Imiquimod refers to 1-isobutyl-1H-imidazo[4,5- c]quinolin-4-amine (CAS No.99011-02-6), or a pharmaceutically acceptable salt thereof, which is a Toll-like receptor 7 (TLR7) agonist.
  • NLG-919 refers to 1-cyclohexyl-2-(5H-imidazo[5,1- a]isoindol-5-yl)ethan-1-ol (CAS No.1402836-58-1), or a pharmaceutically acceptable salt thereof, which is an indoleamine 2,3-dioxygenase (IDO) inhibitor.
  • L-NMMA refers to N 5 - [imino(methylamino)methyl]-L-ornithine (L-N G -monomethyl Arginine) (CAS No.53308-83-1, acetate salt), or a pharmaceutically acceptable salt thereof, which is a nitric oxide synthase (NOS) inhibitor.
  • Nor-NOHA refers to (S)-2-amino-4-(3-hydroxyguanidino)butanoic acid (N ⁇ -Hydroxy-nor- L-arginine) (CAS No.291758-32-2, dichloride salt), or a pharmaceutically acceptable salt thereof, which is an arginase inhibitor.
  • CA-170 refers to (((S)-3-amino-1-(3-((R)-1-amino-2-hydroxyethyl)- 1,2,4-oxadiazol-5-yl)-3-oxopropyl)carbamoyl)-L-threonine (CAS No.1673534-76-3), or a pharmaceutically acceptable salt thereof, which is a programmed death-ligand 1 (PD-L1) inhibitor.
  • PD-L1 programmed death-ligand 1
  • Triciribine phosphate refers to ((2S,3R,4S,5S)-5-(3-amino-5-methyl-1,4,5,6,8-pentaazaacenaphthylen- 1(5H)-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl dihydrogen phosphate (CAS No.61966-08-3), or a pharmaceutically acceptable salt thereof, which is a protein kinase B inhibitor.
  • AB-680 refers to (((((2R,3S,4R,5R)-5-(6-chloro-4-(((S)-1-(2-fluorophenyl)ethyl)amino)-1H-pyrazolo[3,4-b]pyridin-1- yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid (CAS No.2105904-82-1), or a pharmaceutically acceptable salt thereof, which is a cluster of differentiation 73 (CD73) inhibitor.2′,3′-cGAMP (2′,3′-cyclic GMP-AMP) refers to 2-amino-9- ((5R,7R,8R,12aR,14R,15R,15aS,16R)-14-(6-amino-9H-purin-9-yl)-2,10,15,16-tetrahydroxy-2,10- dioxidooctahydro-12H-5
  • the conjugated agent is a radical of an agent selected from LLL12, imiquimod, and NLG-919. In some embodiments, the conjugated agent is a radical of LLL12. In some embodiments, the conjugated agent is a radical of imiquimod. In some embodiments, the conjugated agent is a radical of NLG-919. In some embodiments, the conjugated agent is a radical of an agent selected from L-NMMA, nor-NOHA, and CA-170. In some embodiments, the conjugated agent is a radical of L-NMMA. In some embodiments, the conjugated agent is a radical of nor-NOHA. In some embodiments, the conjugated agent is a radical of CA-170.
  • the conjugated agent is a radical of an agent selected from triciribine phosphate, AB-680, and 2′,3′-cGAMP. In some embodiments, the conjugated agent is a radical of triciribine phosphate. In some embodiments, the conjugated agent is a radical of AB-680. In some embodiments, the conjugated agent is a radical of
  • the conjugated agent is a radical of an agent selected from LLL12 and LLL12-Hydrazone.
  • at least one instance of X is a radical of an agent selected from LLL12, imiquimod, NLG-919, L-NMMA, nor-NOHA, CA-170, triciribine phosphate, AB-680, and 2′,3′- cGAMP.
  • at least one instance of X is a radical of an agent selected from LLL12, imiquimod, and NLG-919.
  • At least one instance of X is a radical of LLL12. In some embodiments, at least one instance of X is a radical of imiquimod. In some embodiments, at least one instance of X is a radical of NLG-919. In some embodiments, at least one instance of X is a radical of an agent selected from L-NMMA, nor-NOHA, and CA-170. In some embodiments, at least one instance of X is a radical of L-NMMA. In some embodiments, at least one instance of X is a radical of nor-NOHA. In some embodiments, at least one instance of X is a radical of CA-170.
  • At least one instance of X is a radical of an agent selected from triciribine phosphate, AB-680, and 2′,3′-cGAMP. In some embodiments, at least one instance of X is a radical of triciribine phosphate. In some embodiments, at least one instance of X is a radical of AB- 680. In some embodiments, at least one instance of X is a radical of 2′,3′-cGAMP.
  • At least one instance of X is a radical of an agent selected from LLL12, imiquimod, NLG-919, L-NMMA, nor-NOHA, CA-170, triciribine phosphate, AB-680, 2′,3′-cGAMP, and LLL12-Hydrazone.
  • at least one instance of X is a radical of an agent selected from LLL12 and LLL12-Hydrazone.
  • at least one instance of X is a radical of LLL12-Hydrazone.
  • each instance of X is independently a radical of an agent selected from LLL12, imiquimod, NLG-919, L-NMMA, nor-NOHA, CA-170, triciribine phosphate, AB-680, and 2′,3′-cGAMP.
  • each instance of X is independently a radical of an agent selected from LLL12, imiquimod, and NLG-919.
  • each instance of X is independently a radical of LLL12.
  • each instance of X is independently a radical of imiquimod.
  • each instance of X is independently a radical of NLG- 919.
  • each instance of X is independently a radical of an agent selected from L- NMMA, nor-NOHA, and CA-170. In some embodiments, each instance of X is independently a radical of L-NMMA. In some embodiments, each instance of X is independently a radical of nor- NOHA. In some embodiments, each instance of X is independently a radical of CA-170. In some embodiments, each instance of X is independently a radical of an agent selected from triciribine phosphate, AB-680, and 2′,3′-cGAMP. In some embodiments, each instance of X is independently a radical of triciribine phosphate. In some embodiments, each instance of X is independently a radical of AB-680.
  • each instance of X is independently a radical of 2′,3′-cGAMP.
  • each instance of X is independently a radical of an agent selected from LLL12, imiquimod, NLG-919, L-NMMA, nor-NOHA, CA-170, triciribine phosphate, AB-680, 2′,3′- cGAMP, and LLL12-Hydrazone.
  • each instance of X is independently a radical
  • the agent is a hydrophobic agent, a cationic agent, or an anionic agent. In some embodiments, the agent is a hydrophobic agent. In some embodiments, the hydrophobic agent has a logP value of greater than 0. In some embodiments, the hydrophobic agent is selected from LLL12, imiquimod, and NLG-919, and pharmaceutically acceptable salts thereof. [0238] In some embodiments, the agent is a cationic agent.
  • the cationic agent is positively charged at physiological pH.
  • the cationic agent is selected from L- NMMA, nor-NOHA, and CA-170, and pharmaceutically acceptable salts thereof.
  • the agent is an anionic agent.
  • the anionic agent is negatively charged at physiological pH.
  • the anionic agent is selected from triciribine phosphate, AB-680, and 2′,3′-cGAMP, and pharmaceutically acceptable salts thereof.
  • the agent is a STAT3 inhibitor, a TLR7 agonist, an IDO inhibitor, an NOS inhibitor, an arginase inhibitor, a PD-L1 inhibitor, a protein kinase B inhibitor, a CD73 inhibitor, or a STING agonist.
  • the agent is a STAT3 inhibitor, a TLR7 agonist, or an IDO inhibitor.
  • the agent is a STAT3 inhibitor.
  • the agent is a TLR7 agonist.
  • the agent is an IDO inhibitor.
  • the agent is an NOS inhibitor, an arginase inhibitor, a PD-L1 inhibitor.
  • the agent is an NOS inhibitor. In some embodiments, the agent is an arginase inhibitor. In some embodiments, the agent is a PD-L1 inhibitor. In some embodiments, the agent is a protein kinase B inhibitor, a CD73 inhibitor, or a STING agonist. In some embodiments, the agent is a protein kinase B inhibitor. In some embodiments, the agent is a CD73 inhibitor. In some embodiments, the agent is a STING agonist. [0241] In some embodiments, the STAT3 inhibitor is LLL12. In some embodiments, the STAT3 inhibitor is LLL12-Hydrazone. In some embodiments, the TLR7 agonist is imiquimod.
  • the IDO inhibitor is NLG-919.
  • the NOS inhibitor is L-NMMA.
  • the arginase inhibitor is nor-NOHA.
  • the PD-L1 inhibitor is CA-170.
  • the protein kinase B inhibitor is triciribine phosphate.
  • the CD73 inhibitor is AB-680.
  • the STING agonist is 2′,3′- cGAMP.
  • At least one instance of X is selected from formulae (ii), (iii), (iv), (v), (vi), (vii), (viii)
  • At least one instance of X is selected from formulae (ii), (iii), (iv), (v), (vi), (vii), (viii) [0244] In some embodiments, at least one instance of X is selected from formulae (ii), (iii), and (iv). In some embodiments, at least one instance of X is of formula (ii). In some embodiments, at least one instance of X is of formula (iii). In some embodiments, at least one instance of X is of formula (iv). In some embodiments, at least one instance of X is selected from formulae (v), (vi), and (vii).
  • At least one instance of X is of formula (v). In some embodiments, at least one instance of X is of formula (vi). In some embodiments, at least one instance of X is of formula (vii). In some embodiments, at least one instance of X is selected from formulae (viii), (ix), and (x). In some embodiments, at least one instance of X is of formula (viii). In some embodiments, at least one
  • U1197.70243WO00 78/193 #13587456v2 instance of X is of formula (ix). In some embodiments, at least one instance of X is of formula (x). In some embodiments, at least one instance of X is selected from formulae (ii) and (xi). In some embodiments, at least one instance of X is of formula (xi). [0245] In some embodiments, each instance of X is independently selected from formulae (ii), (iii), (iv), (v), (vi), (vii), (viii), (ix), and (x). In some embodiments, each instance of X is independently selected from formulae (ii), (iii), and (iv).
  • each instance of X is independently of formula (ii). In some embodiments, each instance of X is independently of formula (iii). In some embodiments, each instance of X is independently of formula (iv). In some embodiments, each instance of X is independently selected from formulae (v), (vi), and (vii). In some embodiments, each instance of X is independently of formula (v). In some embodiments, each instance of X is independently of formula (vi). In some embodiments, each instance of X is independently of formula (vii). In some embodiments, each instance of X is independently selected from formulae (viii), (ix), and (x). In some embodiments, each instance of X is independently of formula (viii).
  • each instance of X is independently of formula (ix). In some embodiments, each instance of X is independently of formula (x). In some embodiments, each instance of X is independently selected from formulae (ii) and (xi). In some embodiments, each instance of X is independently of formula (xi). [0246] In some embodiments, the compound of Formula (I) or Formula (I′) is selected from
  • the compound of Formula (I) or Formula (I′) is selected from Formulae
  • the compound of Formula (I) or Formula (I′) is selected from Formulae (II), (III), (IV), and (XI).
  • the compound of Formula (I) or Formula (I′) is selected from Formulae (II), (III), and (IV).
  • the compound of Formula (I) or Formula (I′) is of Formula (II).
  • the compound of Formula (I) or Formula (I′) is of Formula (III).
  • the compound of Formula (I) or Formula (I′) is of Formula (IV).
  • the compound of Formula (I) or Formula (I′) is selected from Formulae (V), (VI), and (VII).
  • the compound of Formula (I) or Formula (I′) is
  • the compound of Formula (I) or Formula (I′) is of Formula (VI). In some embodiments, the compound of Formula (I) or Formula (I′) is of Formula (VII). In some embodiments, the compound of Formula (I) or Formula (I′) is selected from Formulae (VIII), (IX), and (X). In some embodiments, the compound of Formula (I) or Formula (I′) is of Formula (VIII). In some embodiments, the compound of Formula (I) or Formula (I′) is of Formula (IX). In some embodiments, the compound of Formula (I) or Formula (I′) is of Formula (X).
  • the compound of Formula (I) or Formula (I′) is selected from Formulae (II) and (XI). In some embodiments, the compound of Formula (I) or Formula (I′) is of Formula (XI). [0249] In some embodiments, the compound of Formula (I) is selected from Formulae (II), (III), (IV), and (XI). In some embodiments, the compound of Formula (I) is selected from Formulae (II), (III), and (IV). In some embodiments, the compound of Formula (I) is of Formula (II). In some embodiments, the compound of Formula (I) is of Formula (III). In some embodiments, the compound of Formula (I) is of Formula (IV).
  • the compound of Formula (I) is selected from Formulae (V), (VI), and (VII). In some embodiments, the compound of Formula (I) is of Formula (V). In some embodiments, the compound of Formula (I) is of Formula (VI). In some embodiments, the compound of Formula (I) is of Formula (VII). In some embodiments, the compound of Formula (I) is selected from Formulae (VIII), (IX), and (X). In some embodiments, the compound of Formula (I) is of Formula (VIII). In some embodiments, the compound of Formula (I) is of Formula (IX). In some embodiments, the compound of Formula (I) is of Formula (X).
  • the compound of Formula (I) is selected from Formulae (II) and (XI). In some embodiments, the compound of Formula (I) is of Formula (XI). [0250] In some embodiments, the compound of Formula (I′) is selected from Formulae (II), (III), (IV), and (XI). In some embodiments, the compound of Formula (I′) is selected from Formulae (II), (III), and (IV). In some embodiments, the compound of Formula (I′) is of Formula (II). In some embodiments, the compound of Formula (I′) is of Formula (III). In some embodiments, the compound of Formula (I′) is of Formula (IV).
  • the compound of Formula (I′) is selected from Formulae (V), (VI), and (VII). In some embodiments, the compound of Formula (I′) is of Formula (V). In some embodiments, the compound of Formula (I′) is of Formula (VI). In some embodiments, the compound of Formula (I′) is of Formula (VII). In some embodiments, the compound of Formula (I′) is selected from Formulae (VIII), (IX), and (X). In some embodiments, the compound of Formula (I′) is of Formula (VIII). In some embodiments, the compound of Formula (I′) is of Formula (IX). In some embodiments, the compound of Formula (I′) is of Formula (X).
  • the compound of Formula (I′) is selected from Formulae (II) and (XI). In some embodiments, the compound of Formula (I′) is of Formula (XI). [0251] In some embodiments, the compound of Formula (I) can be characterized in terms of mass percentage (e.g., % by mass (m/m)) of X (e.g., of conjugated agent). In some embodiments, mass percentage refers to a molecular weight (Da) percentage of X (e.g., of conjugated agent) in the
  • mass percentage can be determined by the general formula of: (X (e.g., conjugated agent) M W ) / (compound of Formula (I) M W ) ⁇ 100.
  • (X (e.g., conjugated agent) M W ) can be determined by calculating or approximating the molecular weight of X (e.g., of a conjugated agent) as a single molecule or compound (conjugated or unconjugated), and multiplying this value by the number of terminal sites at which X (e.g., of conjugated agent) is present in the compound of Formula (I) (e.g., multiplying by p).
  • (X (e.g., conjugated agent) M W ) can be determined by calculating or approximating the sum of the atomic mass of all atoms which form X (e.g., conjugated agent) in the compound of Formula (I).
  • the value for (X (e.g., conjugated agent) MW) can be taken as a fraction of total molecular weight of the compound of Formula (I) (compound of Formula (I) MW), and multiplied by 100 to provide a mass percentage.
  • mass percentage can be determined by experimental or empirical means. For example, in some embodiments, mass percentage can be determined using proton nuclear magnetic resonance ( 1 H NMR) or other analytical methods known in the art.
  • the compound of Formula (I′) can be characterized in terms of mass percentage (e.g., % by mass (m/m)) of X. In some embodiments, mass percentage refers to a molecular weight (Da) percentage of X in the compound of Formula (I′).
  • mass percentage can be determined by the general formula of: (X MW) / (compound of Formula (I′) MW) ⁇ 100.
  • (X MW) can be determined by calculating or approximating the molecular weight of X as a single molecule or compound (conjugated or unconjugated), and multiplying this value by the number of terminal sites at which X is present in the compound of Formula (I′) (e.g., multiplying by p).
  • (X MW) can be determined by calculating or approximating the sum of the atomic mass of all atoms which form X in the compound of Formula (I′).
  • the value for (X MW) can be taken as a fraction of total molecular weight of the compound of Formula (I′) (compound of Formula (I′) MW), and multiplied by 100 to provide a mass percentage.
  • mass percentage can be determined by experimental or empirical means. For example, in some embodiments, mass percentage can be determined using proton nuclear magnetic resonance ( 1 H NMR) or other analytical methods known in the art.
  • the compound of Formula (I) comprises between about 1% and about 20%, about 1% and about 15%, about 1% and about 14%, about 1% and about 13%, about 1% and about 12%, about 1% and about 11%, about 1% and about 10%, about 1% and about 9%, about 1% and about 8%, about 1% and about 7%, about 1% and about 6%, about 1% and about 5%, about 2% and about 15%, about 2% and about 10%, about 3% and about 15%, about 3% and about 10%, about 4% and about 15%, about 5% and about 10%, about 6% and about 15%, about 7% and about 10%, about 8% and about 15%, or about 9% and about 10% by mass (m/m) of X (e.g., of conjugated agent).
  • the compound of Formula (I) comprises between about 1% and about 20% by mass of X (e.g., of conjugated agent).
  • the compound of Formula (I) comprises
  • U1197.70243WO00 83/193 #13587456v2 between about 1% and about 10% by mass of X (e.g., of conjugated agent).
  • the compound of Formula (I) comprises between about 5% and about 10% by mass of X (e.g., of conjugated agent).
  • the compound of Formula (I′) comprises between about 1% and about 20%, about 1% and about 15%, about 1% and about 14%, about 1% and about 13%, about 1% and about 12%, about 1% and about 11%, about 1% and about 10%, about 1% and about 9%, about 1% and about 8%, about 1% and about 7%, about 1% and about 6%, about 1% and about 5%, about 2% and about 15%, about 2% and about 10%, about 3% and about 15%, about 3% and about 10%, about 4% and about 15%, about 5% and about 10%, about 6% and about 15%, about 7% and about 10%, about 8% and about 15%, or about 9% and about 10% by mass (m/m) of X.
  • the compound of Formula (I′) comprises between about 1% and about 20% by mass of X. In some embodiments, the compound of Formula (I′) comprises between about 1% and about 10% by mass of X. In some embodiments, the compound of Formula (I′) comprises between about 5% and about 10% by mass of X.
  • the compound of Formula (II) comprises between about 1% and about 20%, about 1% and about 15%, about 1% and about 14%, about 1% and about 13%, about 1% and about 12%, about 1% and about 11%, about 1% and about 10%, about 1% and about 9%, about 1% and about 8%, about 1% and about 7%, about 1% and about 6%, about 1% and about 5%, about 2% and about 15%, about 2% and about 10%, about 3% and about 15%, about 3% and about 10%, about 4% and about 15%, about 5% and about 10%, about 6% and about 15%, about 7% and about 10%, about 8% and about 15%, or about 9% and about 10% by mass (m/m) of formula (ii).
  • the compound of Formula (II) comprises between about 1% and about 20% by mass of formula (ii). In some embodiments, the compound of Formula (II) comprises between about 1% and about 10% by mass of formula (ii). In some embodiments, the compound of Formula (II) comprises between about 5% and about 10% by mass of formula (ii).
  • the compound of Formula (III) comprises between about 1% and about 20%, about 1% and about 15%, about 1% and about 14%, about 1% and about 13%, about 1% and about 12%, about 1% and about 11%, about 1% and about 10%, about 1% and about 9%, about 1% and about 8%, about 1% and about 7%, about 1% and about 6%, about 1% and about 5%, about 2% and about 15%, about 2% and about 10%, about 3% and about 15%, about 3% and about 10%, about 4% and about 15%, about 5% and about 10%, about 6% and about 15%, about 7% and about 10%, about 8% and about 15%, or about 9% and about 10% by mass (m/m) of formula (iii).
  • the compound of Formula (III) comprises between about 1% and about 20% by mass of formula (iii). In some embodiments, the compound of Formula (III) comprises between about 1% and about 10% by mass of formula (iii). In some embodiments, the compound of Formula (III) comprises between about 5% and about 10% by mass of formula (iii).
  • the compound of Formula (IV) comprises between about 1% and about 20%, about 1% and about 15%, about 1% and about 14%, about 1% and about 13%, about 1% and about 12%, about 1% and about 11%, about 1% and about 10%, about 1% and about 9%, about 1% and about 8%, about 1% and about 7%, about 1% and about 6%, about 1% and about 5%, about 2% and about 15%, about 2% and about 10%, about 3% and about 15%, about 3% and about 10%, about 4% and about 15%, about 5% and about 10%, about 6% and about 15%, about 7% and about 10%, about 8% and about 15%, or about 9% and about 10% by mass (m/m) of formula (iv).
  • the compound of Formula (IV) comprises between about 1% and about 20% by mass of formula (iv). In some embodiments, the compound of Formula (IV) comprises between about 1% and about 10% by mass of formula (iv). In some embodiments, the compound of Formula (IV) comprises between about 5% and about 10% by mass of formula (iv).
  • the compound of Formula (V) comprises between about 1% and about 20%, about 1% and about 15%, about 1% and about 14%, about 1% and about 13%, about 1% and about 12%, about 1% and about 11%, about 1% and about 10%, about 1% and about 9%, about 1% and about 8%, about 1% and about 7%, about 1% and about 6%, about 1% and about 5%, about 2% and about 15%, about 2% and about 10%, about 3% and about 15%, about 3% and about 10%, about 4% and about 15%, about 5% and about 10%, about 6% and about 15%, about 7% and about 10%, about 8% and about 15%, or about 9% and about 10% by mass (m/m) of formula (v).
  • the compound of Formula (V) comprises between about 1% and about 20% by mass of formula (v). In some embodiments, the compound of Formula (V) comprises between about 1% and about 10% by mass of formula (v). In some embodiments, the compound of Formula (V) comprises between about 5% and about 10% by mass of formula (v).
  • the compound of Formula (VI) comprises between about 1% and about 20%, about 1% and about 15%, about 1% and about 14%, about 1% and about 13%, about 1% and about 12%, about 1% and about 11%, about 1% and about 10%, about 1% and about 9%, about 1% and about 8%, about 1% and about 7%, about 1% and about 6%, about 1% and about 5%, about 2% and about 15%, about 2% and about 10%, about 3% and about 15%, about 3% and about 10%, about 4% and about 15%, about 5% and about 10%, about 6% and about 15%, about 7% and about 10%, about 8% and about 15%, or about 9% and about 10% by mass (m/m) of formula (vi).
  • the compound of Formula (VI) comprises between about 1% and about 20% by mass of formula (vi). In some embodiments, the compound of Formula (VI) comprises between about 1% and about 10% by mass of formula (vi). In some embodiments, the compound of Formula (VI) comprises between about 5% and about 10% by mass of formula (vi).
  • the compound of Formula (VII) comprises between about 1% and about 20%, about 1% and about 15%, about 1% and about 14%, about 1% and about 13%, about 1% and about 12%, about 1% and about 11%, about 1% and about 10%, about 1% and about 9%, about 1% and about 8%, about 1% and about 7%, about 1% and about 6%, about 1% and about 5%, about
  • the compound of Formula (VII) comprises between about 1% and about 20% by mass of formula (vii). In some embodiments, the compound of Formula (VII) comprises between about 1% and about 10% by mass of formula (vii). In some embodiments, the compound of Formula (VII) comprises between about 5% and about 10% by mass of formula (vii).
  • the compound of Formula (VIII) comprises between about 1% and about 20%, about 1% and about 15%, about 1% and about 14%, about 1% and about 13%, about 1% and about 12%, about 1% and about 11%, about 1% and about 10%, about 1% and about 9%, about 1% and about 8%, about 1% and about 7%, about 1% and about 6%, about 1% and about 5%, about 2% and about 15%, about 2% and about 10%, about 3% and about 15%, about 3% and about 10%, about 4% and about 15%, about 5% and about 10%, about 6% and about 15%, about 7% and about 10%, about 8% and about 15%, or about 9% and about 10% by mass (m/m) of formula (viii).
  • the compound of Formula (VIII) comprises between about 1% and about 20% by mass of formula (viii). In some embodiments, the compound of Formula (VIII) comprises between about 1% and about 10% by mass of formula (viii). In some embodiments, the compound of Formula (VIII) comprises between about 5% and about 10% by mass of formula (viii).
  • the compound of Formula (IX) comprises between about 1% and about 20%, about 1% and about 15%, about 1% and about 14%, about 1% and about 13%, about 1% and about 12%, about 1% and about 11%, about 1% and about 10%, about 1% and about 9%, about 1% and about 8%, about 1% and about 7%, about 1% and about 6%, about 1% and about 5%, about 2% and about 15%, about 2% and about 10%, about 3% and about 15%, about 3% and about 10%, about 4% and about 15%, about 5% and about 10%, about 6% and about 15%, about 7% and about 10%, about 8% and about 15%, or about 9% and about 10% by mass (m/m) of formula (ix).
  • the compound of Formula (IX) comprises between about 1% and about 20% by mass of formula (ix). In some embodiments, the compound of Formula (IX) comprises between about 1% and about 10% by mass of formula (ix). In some embodiments, the compound of Formula (IX) comprises between about 5% and about 10% by mass of formula (ix).
  • the compound of Formula (X) comprises between about 1% and about 20%, about 1% and about 15%, about 1% and about 14%, about 1% and about 13%, about 1% and about 12%, about 1% and about 11%, about 1% and about 10%, about 1% and about 9%, about 1% and about 8%, about 1% and about 7%, about 1% and about 6%, about 1% and about 5%, about 2% and about 15%, about 2% and about 10%, about 3% and about 15%, about 3% and about 10%, about 4% and about 15%, about 5% and about 10%, about 6% and about 15%, about 7% and about 10%, about 8% and about 15%, or about 9% and about 10% by mass (m/m) of formula (x).
  • the compound of Formula (X) comprises between about 1% and about 20% by mass of
  • the compound of Formula (X) comprises between about 1% and about 10% by mass of formula (x). In some embodiments, the compound of Formula (X) comprises between about 5% and about 10% by mass of formula (x).
  • the compound of Formula (XI) comprises between about 1% and about 20%, about 1% and about 15%, about 1% and about 14%, about 1% and about 13%, about 1% and about 12%, about 1% and about 11%, about 1% and about 10%, about 1% and about 9%, about 1% and about 8%, about 1% and about 7%, about 1% and about 6%, about 1% and about 5%, about 2% and about 15%, about 2% and about 10%, about 3% and about 15%, about 3% and about 10%, about 4% and about 15%, about 5% and about 10%, about 6% and about 15%, about 7% and about 10%, about 8% and about 15%, or about 9% and about 10% by mass (m/m) of formula (xi).
  • the compound of Formula (XI) comprises between about 1% and about 20% by mass of formula (xi). In some embodiments, the compound of Formula (XI) comprises between about 1% and about 10% by mass of formula (xi). In some embodiments, the compound of Formula (XI) comprises between about 5% and about 10% by mass of formula (xi).
  • At least one instance of Y is –OH. In some embodiments, each instance of Y is independently –OH. [0268] In some embodiments, at least one instance of Y is –OR A , wherein R A is hydrogen, optionally substituted alkyl, optionally substituted alkynyl, or optionally substituted heteroalkyl. In some embodiments, at least one instance of Y is –OR A , wherein R A is hydrogen or optionally substituted alkyl. In some embodiments, at least one instance of Y is –OR A , wherein R A is hydrogen or optionally substituted alkynyl.
  • At least one instance of Y is –OR A , wherein R A is hydrogen or optionally substituted heteroalkyl. In some embodiments, at least one instance of Y is – OR A , wherein R A is hydrogen, optionally substituted alkyl, or optionally substituted alkynyl. In some embodiments, at least one instance of Y is –OR A , wherein R A is hydrogen, optionally substituted alkyl, or optionally substituted heteroalkyl. In some embodiments, at least one instance of Y is –OR A , wherein R A is hydrogen, optionally substituted alkynyl, or optionally substituted heteroalkyl.
  • each instance of Y is independently –OR A , wherein R A is hydrogen, optionally substituted alkyl, optionally substituted alkynyl, or optionally substituted heteroalkyl. In some embodiments, each instance of Y is independently –OR A , wherein R A is hydrogen or optionally substituted alkyl. In some embodiments, each instance of Y is independently –OR A , wherein R A is hydrogen or optionally substituted alkynyl. In some embodiments, each instance of Y is
  • each instance of Y is independently –OR A , wherein R A is hydrogen, optionally substituted alkyl, or optionally substituted alkynyl. In some embodiments, each instance of Y is independently –OR A , wherein R A is hydrogen, optionally substituted alkyl, or optionally substituted heteroalkyl. In some embodiments, each instance of Y is independently –OR A , wherein R A is hydrogen, optionally substituted alkynyl, or optionally substituted heteroalkyl.
  • At least one instance of Y is –OR A , wherein R A is optionally substituted alkyl, optionally substituted alkynyl, or optionally substituted heteroalkyl. In some embodiments, at least one instance of Y is –OR A , wherein R A is optionally substituted alkyl or optionally substituted alkynyl. In some embodiments, at least one instance of Y is –OR A , wherein R A is optionally substituted alkyl or optionally substituted heteroalkyl. In some embodiments, at least one instance of Y is –OR A , wherein R A is optionally substituted alkynyl or optionally substituted heteroalkyl.
  • At least one instance of Y is –OR A , wherein R A is hydrogen, optionally substituted C1-10 alkyl, optionally substituted C1-10 alkynyl, or optionally substituted C1-10 heteroalkyl. In some embodiments, at least one instance of Y is –OR A , wherein R A is hydrogen, optionally substituted linear C1-10 alkyl, optionally substituted linear C1-10 alkynyl, or optionally substituted linear C1-10 heteroalkyl.
  • At least one instance of Y is –OR A , wherein R A is hydrogen, optionally substituted C1-6 alkyl, optionally substituted C1-6 alkynyl, or optionally substituted C1-6 heteroalkyl. In some embodiments, at least one instance of Y is –OR A , wherein R A is hydrogen, optionally substituted linear C1-6 alkyl, optionally substituted linear C1-6 alkynyl, or optionally substituted linear C1-6 heteroalkyl.
  • at least one instance of Y is –OR A , wherein R A is hydrogen, linear C 1-6 alkyl,
  • U1197.70243WO00 88/193 #13587456v2 linear C 1-6 alkynyl, or linear C 1-6 heteroalkyl, wherein the alkyl, alkynyl, or heteroalkyl is substituted with an oxo group ( O) and/or an amino group (–NH 2 ).
  • each instance of Y is independently –OR A , wherein R A is hydrogen, optionally substituted C 1-10 alkyl, optionally substituted C 1-10 alkynyl, or optionally substituted C 1-10 heteroalkyl. In some embodiments, each instance of Y is independently –OR A , wherein R A is hydrogen, optionally substituted linear C1-10 alkyl, optionally substituted linear C1-10 alkynyl, or optionally substituted linear C1-10 heteroalkyl. In some embodiments, each instance of Y is independently –OR A , wherein R A is hydrogen, optionally substituted C1-6 alkyl, optionally substituted C1-6 alkynyl, or optionally substituted C1-6 heteroalkyl.
  • each instance of Y is independently –OR A , wherein R A is hydrogen, optionally substituted linear C1-6 alkyl, optionally substituted linear C1-6 alkynyl, or optionally substituted linear C1-6 heteroalkyl.
  • At least one instance of Y is –OH, or [0276] In some embodiments, at least one instance of Y is –N(R A ) 2 . In some embodiments, each instance of Y is independently –N(R A ) 2 . In some embodiments, at least one instance of Y is –NHR A . In some embodiments, each instance of Y is independently –NHR A . In some embodiments, at least one instance of Y is –NH 2 . In some embodiments, each instance of Y is independently –NH 2 .
  • each instance of R A is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl.
  • at least one instance of R A is hydrogen, optionally substituted alkyl, optionally substituted alkynyl, or optionally substituted heteroalkyl. In some embodiments, at least one instance of R A is hydrogen or optionally substituted alkyl.
  • At least one instance of R A is hydrogen or optionally substituted alkynyl. In some embodiments, at least one instance of R A is hydrogen or optionally substituted heteroalkyl. In some embodiments, at least one instance of R A is hydrogen, optionally substituted alkyl, or optionally substituted alkynyl. In some embodiments, at least one instance of R A is hydrogen, optionally substituted alkyl, or optionally substituted heteroalkyl. In some embodiments, at least one instance of R A is hydrogen, optionally substituted alkynyl, or optionally substituted heteroalkyl.
  • each instance of R A is independently hydrogen, optionally substituted alkyl, optionally substituted alkynyl, or optionally substituted heteroalkyl. In some embodiments, each instance of R A is independently hydrogen or optionally substituted alkyl. In some embodiments, each instance of R A is independently hydrogen or optionally substituted alkynyl. In some embodiments, each instance of R A is independently hydrogen or optionally substituted heteroalkyl. In some
  • each instance of R A is independently hydrogen, optionally substituted alkyl, or optionally substituted alkynyl. In some embodiments, each instance of R A is independently hydrogen, optionally substituted alkyl, or optionally substituted heteroalkyl. In some embodiments, each instance of R A is independently hydrogen, optionally substituted alkynyl, or optionally substituted heteroalkyl. [0281] In some embodiments, at least one instance of R A is optionally substituted alkyl, optionally substituted alkynyl, or optionally substituted heteroalkyl.
  • At least one instance of R A is hydrogen, optionally substituted C1-10 alkyl, optionally substituted C1-10 alkynyl, or optionally substituted C1-10 heteroalkyl. In some embodiments, at least one instance of R A is hydrogen, optionally substituted linear C1-10 alkyl, optionally substituted linear C1-10 alkynyl, or optionally substituted linear C1-10 heteroalkyl. In some embodiments, at least one instance of R A is hydrogen, optionally substituted C1-6 alkyl, optionally substituted C1-6 alkynyl, or optionally substituted C1-6 heteroalkyl.
  • At least one instance of R A is hydrogen, optionally substituted linear C1-6 alkyl, optionally substituted linear C1-6 alkynyl, or optionally substituted linear C1-6 heteroalkyl.
  • each instance of R A is independently hydrogen, optionally substituted C 1-10 alkyl, optionally substituted C 1-10 alkynyl, or optionally substituted C 1-10 heteroalkyl.
  • each instance of R A is independently hydrogen, optionally substituted linear C 1-10 alkyl, optionally substituted linear C 1-10 alkynyl, or optionally substituted linear C 1-10 heteroalkyl.
  • each instance of R A is independently hydrogen, optionally substituted C 1-6 alkyl, optionally substituted C 1-6 alkynyl, or optionally substituted C 1-6 heteroalkyl. In some embodiments, each instance of R A is independently hydrogen, optionally substituted linear C 1-6 alkyl, optionally substituted linear C 1-6 alkynyl, or optionally substituted linear C 1-6 heteroalkyl.
  • At least one instance of R A is hydrogen, or [0287] In some embodiments, at least one instance of R A is of formula: (i), wherein R A1 is optionally substituted alkyl, optionally substituted alkynyl, or optionally substituted heteroalkyl. In
  • At least one instance of R A is of formula (i), wherein R A1 is optionally substituted C 1-10 alkyl, optionally substituted C 1-10 alkynyl, or optionally substituted C 1-10 heteroalkyl. In some embodiments, at least one instance of R A is of formula (i), wherein R A1 is optionally substituted linear C 1-10 alkyl, optionally substituted linear C 1-10 alkynyl, or optionally substituted linear C 1-10 heteroalkyl.
  • At least one instance of R A is of formula (i), wherein R A1 is optionally substituted C 1-6 alkyl, optionally substituted C 1-6 alkynyl, or optionally substituted C 1-6 heteroalkyl. In some embodiments, at least one instance of R A is of formula (i), wherein R A1 is optionally substituted linear C1-6 alkyl, optionally substituted linear C1-6 alkynyl, or optionally substituted linear C1-6 heteroalkyl. In some embodiments, at least one instance of R A is of formula (i), wherein R A1 is [0288] In some embodiments, at least one instance of R A is hydrogen. In some embodiments, each instance of R A is hydrogen.
  • At least one instance of R A is optionally substituted alkenyl. In some embodiments, at least one instance of R A is optionally substituted C1-10 alkenyl. In some embodiments, at least one instance of R A is optionally substituted linear C 1-10 alkenyl. In some embodiments, at least one instance of R A is optionally substituted C 1-6 alkenyl. In some embodiments, at least one instance of R A is optionally substituted linear C 1-6 alkenyl. In some embodiments, each instance of R A is independently hydrogen or optionally substituted alkenyl. In some embodiments, each instance of R A is independently hydrogen or optionally substituted C 1-10 alkenyl.
  • each instance of R A is independently hydrogen or optionally substituted linear C 1-10 alkenyl. In some embodiments, each instance of R A is independently hydrogen or optionally substituted C 1-6 alkenyl. In some embodiments, each instance of R A is independently hydrogen or optionally substituted linear C 1-6 alkenyl. [0294] In some embodiments, at least one instance of R A is optionally substituted alkynyl. In some embodiments, at least one instance of R A is optionally substituted C 1-10 alkynyl. In some embodiments, at least one instance of R A is optionally substituted linear C 1-10 alkynyl. In some embodiments, at least one instance of R A is optionally substituted C 1-6 alkynyl.
  • At least one instance of R A is optionally substituted linear C 1-6 alkynyl.
  • At least one instance of R A is optionally substituted linear C1-10 heteroalkenyl. In some embodiments, at least one instance of R A is optionally substituted C1-6 heteroalkenyl. In some embodiments, at least one instance of R A is optionally substituted linear C1-6 heteroalkenyl. In some embodiments, each instance of R A is independently hydrogen or optionally substituted heteroalkenyl. In some embodiments, each instance of R A is independently hydrogen or optionally substituted C1-10 heteroalkenyl. In some embodiments, each instance of R A is independently hydrogen or optionally substituted linear C1-10 heteroalkenyl. In some embodiments, each instance of R A is independently is independently hydrogen or optionally substituted linear C1-10 heteroalkenyl. In some embodiments, each instance of R A is independently
  • each instance of R A is independently hydrogen or optionally substituted linear C 1-6 heteroalkenyl.
  • at least one instance of R A is optionally substituted heteroalkynyl.
  • at least one instance of R A is optionally substituted C 1-10 heteroalkynyl.
  • at least one instance of R A is optionally substituted linear C1-10 heteroalkynyl.
  • at least one instance of R A is optionally substituted C 1-6 heteroalkynyl.
  • At least one instance of R A is optionally substituted linear C 1-6 heteroalkynyl. In some embodiments, each instance of R A is independently hydrogen or optionally substituted heteroalkynyl. In some embodiments, each instance of R A is independently hydrogen or optionally substituted C1-10 heteroalkynyl. In some embodiments, each instance of R A is independently hydrogen or optionally substituted linear C1-10 heteroalkynyl. In some embodiments, each instance of R A is independently hydrogen or optionally substituted C1-6 heteroalkynyl. In some embodiments, each instance of R A is independently hydrogen or optionally substituted linear C1-6 heteroalkynyl. [0304] In some embodiments, at least one instance of R A is optionally substituted carbocyclyl.
  • At least one instance of R A is optionally substituted C3-10 carbocyclyl. In some embodiments, at least one instance of R A is optionally substituted C4-6 carbocyclyl. In some embodiments, each instance of R A is independently hydrogen or optionally substituted carbocyclyl. In some embodiments, each instance of R A is independently hydrogen or optionally substituted C3-10 carbocyclyl. In some embodiments, each instance of R A is independently hydrogen or optionally substituted C4-6 carbocyclyl. [0305] In some embodiments, at least one instance of R A is optionally substituted heterocyclyl. In some embodiments, at least one instance of R A is optionally substituted 3-10 membered heterocyclyl.
  • At least one instance of R A is optionally substituted 4-6 membered heterocyclyl. In some embodiments, each instance of R A is independently hydrogen or optionally substituted heterocyclyl. In some embodiments, each instance of R A is independently hydrogen or optionally substituted 3-10 membered heterocyclyl. In some embodiments, each instance of R A is independently hydrogen or optionally substituted 4-6 membered heterocyclyl. [0306] In some embodiments, at least one instance of R A is optionally substituted aryl. In some embodiments, at least one instance of R A is optionally substituted phenyl. In some embodiments, each instance of R A is independently hydrogen or optionally substituted aryl.
  • each instance of R A is independently hydrogen or optionally substituted phenyl. [0307] In some embodiments, at least one instance of R A is optionally substituted heteroaryl. In some embodiments, at least one instance of R A is optionally substituted 5-10 membered heteroaryl. In some embodiments, at least one instance of R A is optionally substituted 5-6 membered heteroaryl. In some embodiments, at least one instance of R A is optionally substituted 5-6 monocyclic membered heteroaryl. In some embodiments, each instance of R A is independently hydrogen or optionally substituted heteroaryl. In some embodiments, each instance of R A is independently hydrogen or
  • each instance of R A is independently hydrogen or optionally substituted 5-6 membered heteroaryl. In some embodiments, each instance of R A is independently hydrogen or optionally substituted 5-6 monocyclic membered heteroaryl.
  • the compound of Formula (I) comprises an effective number of instances of Y for targeting a specific cell type. In some embodiments, the compound of Formula (I) comprises an effective number of instances of Y for targeting myeloid-derived suppressor cells (MDSCs). In some embodiments, the compound of Formula (I′) comprises an effective number of instances of Y for targeting a specific cell type.
  • the compound of Formula (I′) comprises an effective number of instances of Y for targeting myeloid-derived suppressor cells (MDSCs).
  • the compound of Formula (I) or Formula (I′) is of Formula (I-a): or a pharmaceutically acceptable salt thereof.
  • the compound of Formula (I) is of Formula (I-a), or a pharmaceutically acceptable salt thereof.
  • the compound of Formula (I′) is of Formula (I-a), or a pharmaceutically acceptable salt thereof.
  • the compound of Formula (II) is of Formula (II-a): or a pharmaceutically acceptable salt thereof.
  • the compound of Formula (II) is of Formula (II-b): or a pharmaceutically acceptable salt thereof.
  • the compound of Formula (II) is of Formula (II-c):
  • the compound of Formula (II) is of Formula (II-d) or Formula (II-e): or a pharmaceutically acceptable salt thereof.
  • the compound of Formula (II) is of Formula (II-d-i) or Formula (II-e-
  • the compound of Formula (II) is of formula: , or a pharmaceutically acceptable salt thereof.
  • the compound of Formula (III) is of Formula (III-a): or a pharmaceutically acceptable salt thereof.
  • the compound of Formula (III) is of Formula (III-b):
  • the compound of Formula (III) is of Formula (III-c): or a pharmaceutically acceptable salt thereof.
  • the compound of Formula (IV) is of Formula (IV-a): or a pharmaceutically acceptable salt thereof.
  • the compound of Formula (IV) is of Formula (IV-b): or a pharmaceutically acceptable salt thereof.
  • the compound of Formula (IV) is of Formula (IV-c): or a pharmaceutically acceptable salt thereof.
  • the compound of Formula (V) is of Formula (V-a): or a pharmaceutically acceptable salt thereof.
  • the compound of Formula (V) is of Formula (V-b): or a pharmaceutically acceptable salt thereof.
  • the compound of Formula (V) is of Formula (V-c): or a pharmaceutically acceptable salt thereof.
  • the compound of Formula (VI) is of Formula (VI-a): or a pharmaceutically acceptable salt thereof.
  • the compound of Formula (VI) is of Formula (VI-b): or a pharmaceutically acceptable salt thereof.
  • the compound of Formula (VI) is of Formula (VI-c): or a pharmaceutically acceptable salt thereof.
  • the compound of Formula (VII) is of Formula (VII-a): or a pharmaceutically acceptable salt thereof.
  • the compound of Formula (VII) is of Formula (VII-b)::
  • the compound of Formula (VII) is of Formula (VII-c): or a pharmaceutically acceptable salt thereof.
  • the compound of Formula (VIII) is of Formula (VIII-a): or a pharmaceutically acceptable salt thereof.
  • the compound of Formula (VIII) is of Formula (VIII-b): or a pharmaceutically acceptable salt thereof.
  • the compound of Formula (VIII) is of Formula (VIII-c):
  • the compound of Formula (IX) is of Formula (IX-a): or a pharmaceutically acceptable salt thereof.
  • the compound of Formula (IX) is of Formula (IX-b): or a pharmaceutically acceptable salt thereof.
  • the compound of Formula (IX) is of Formula (IX-c):
  • the compound of Formula (X) is of Formula (X-a): or a pharmaceutically acceptable salt thereof.
  • the compound of Formula (X) is of Formula (X-b): or a pharmaceutically acceptable salt thereof.
  • the compound of Formula (X) is of Formula (X-c): or a pharmaceutically acceptable salt thereof.
  • the compound of Formula (XI) is of Formula (XI-a):
  • the compound of Formula (XI) is of Formula (XI-b): or a pharmaceutically acceptable salt thereof.
  • the compound of Formula (XI) is of Formula (XI-c): or a pharmaceutically acceptable salt thereof.
  • the compound of Formula (XI) is of Formula (XI-d):
  • the compound of Formula (XI) is of Formula (XI-e): or a pharmaceutically acceptable salt thereof.
  • the compound of Formula (XI) is of formula: , or a pharmaceutically acceptable salt thereof.
  • a provided compound (a compound described herein, a compound of the present disclosure) is a compound of any of the formulae disclosed herein (e.g., Formulae (I), (I′), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), (XI)), or a pharmaceutically acceptable salt thereof.
  • a provided compound is a compound of any of the Formulae disclosed herein (e.g., Formulae (I), (I′), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), (XI)), or a salt thereof.
  • a provided compound is a compound of any of the Formulae disclosed herein (e.g., Formulae (I), (I′), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), (XI)).
  • Compositions and Kits [0347]
  • the provided compound is provided in an effective amount in the pharmaceutical composition.
  • the effective amount is an amount effective for modulating a target protein in a subject or a cell, tissue, or biological sample.
  • the composition is formulated as a particle.
  • the composition is formulated as a nanoparticle or microparticle.
  • the particle is a microparticle (i.e., particle having a characteristic dimension of less than about 1 millimeter and at
  • the particle is a nanoparticle (i.e., a particle having a characteristic dimension of less than about 1 micrometer and at least about 1 nanometer, where the characteristic dimension of the particle is the smallest cross-sectional dimension of the particle).
  • the particles described herein may include additional materials such as polymers (e.g., synthetic polymers (e.g., PEG, PLGA) and natural polymers (e.g., phospholipids)).
  • the additional materials are approved by a regulatory agency, such as the U.S.
  • the particles may be prepared using any method known in the art, such as precipitation, milling, spray drying, single and double emulsion solvent evaporation, solvent extraction, phase separation, and simple and complex coacervation.
  • the conditions used in preparing the particles may be altered to yield particles of a desired size or property (e.g., hydrophobicity, hydrophilicity, external morphology, “stickiness”, shape, polydispersity, etc.).
  • the method of preparing the particle and the conditions (e.g., solvent, temperature, concentration, and air flow rate, etc.) used may also depend on the agent being complexed, encapsulated, or mixed, and/or the composition of the matrix.
  • the particles may also be coated.
  • the particles are coated with a targeting agent.
  • the particles are coated with a surface-altering agent.
  • the particles are coated to achieve desirable surface properties (e.g., a particular charge).
  • Pharmaceutical compositions described herein can be prepared by any method known in the art of pharmaceutics. In general, such preparatory methods include bringing the compound described herein (i.e., the “active ingredient”) into association with a carrier or excipient, and/or one or more other accessory ingredients, and then, if necessary and/or desirable, shaping, and/or packaging the product into a desired single- or multi-dose unit.
  • compositions can be prepared, packaged, and/or sold in bulk, as a single unit dose, and/or as a plurality of single unit doses.
  • a “unit dose” is a discrete amount of the pharmaceutical composition comprising a predetermined amount of the active ingredient.
  • the amount of the active ingredient is generally equal to the dosage of the active ingredient which would be administered to a subject and/or a convenient fraction of such a dosage, such as one-half or one-third of such a dosage.
  • Relative amounts of the active ingredient, the pharmaceutically acceptable excipient, and/or any additional ingredients in a pharmaceutical composition described herein will vary, depending upon the identity, size, and/or condition of the subject treated and further depending upon the route by which the composition is to be administered.
  • the composition may comprise between 0.1% and 100% (w/w) active ingredient.
  • Pharmaceutically acceptable excipients used in the manufacture of provided pharmaceutical compositions include inert diluents or fillers, dispersing and/or granulating agents, surface active agents and/or emulsifiers, disintegrating agents, binding agents, preservatives, buffering agents, lubricating agents, and/or oils.
  • Excipients such as cocoa butter and suppository waxes, coloring agents, coating agents, sweetening, flavoring, and perfuming agents may also be present in the composition.
  • Exemplary diluents or fillers include calcium carbonate, sodium carbonate, calcium phosphate, dicalcium phosphate, calcium sulfate, calcium hydrogen phosphate, sodium phosphate lactose, sucrose, cellulose, microcrystalline cellulose, kaolin, mannitol, sorbitol, inositol, sodium chloride, starches (such as dry starch, cornstarch), sugars (such as powdered sugar), calcium trisulfate, carboxymethylcellulose calcium, dextrate, dextrin, dextrose, fructose, lactitol, lactose, magnesium carbonate, magnesium, maltitol, maltodextrin, maltose, sucrose, glucose, mannitol, silicic acid, xylitol,
  • Exemplary granulating and/or dispersing agents include potato starch, corn starch, tapioca starch, sodium starch glycolate, clays, alginic acid, guar gum, citrus pulp, agar, bentonite, cellulose, and wood products, natural sponge, cation-exchange resins, calcium carbonate, silicates, sodium carbonate, cross-linked poly(vinyl-pyrrolidone) (crospovidone), sodium carboxymethyl starch (sodium starch glycolate), carboxymethyl cellulose, cross-linked sodium carboxymethyl cellulose (croscarmellose), methylcellulose, pregelatinized starch (starch 1500), microcrystalline starch, water insoluble starch, calcium carboxymethyl cellulose, magnesium aluminum silicate (Veegum), sodium lauryl sulfate, quaternary ammonium compounds, and mixtures thereof.
  • crospovidone cross-linked poly(vinyl-pyrrolidone)
  • sodium carboxymethyl starch sodium starch glycolate
  • Exemplary surface active agents and/or emulsifiers include natural emulsifiers (e.g., acacia, agar, alginic acid, sodium alginate, tragacanth, chondrux, cholesterol, xanthan, pectin, gelatin, egg yolk, casein, wool fat, cholesterol, wax, and lecithin), colloidal clays (e.g., bentonite (aluminum silicate) and Veegum (magnesium aluminum silicate)), long chain amino acid derivatives, high molecular weight alcohols (e.g., stearyl alcohol, cetyl alcohol, oleyl alcohol, triacetin monostearate, ethylene glycol distearate, glyceryl monostearate, and propylene glycol monostearate, polyvinyl alcohol), carbomers (e.g., carboxy polymethylene, polyacrylic acid, acrylic acid polymer, and carboxyvinyl polymer), carrageenan, cell
  • Exemplary disintegrating agents or disintegrants include agar, algin, alginic acid, sodium alginate, silicates, sodium carbonate, calcium carbonate, carboxymethylcellulose, cellulose, clay, colloidal silicon dioxide, croscarmellose sodium, crospovidone, rubber, magnesium silicate, methylcellulose, potassium krillin, hydroxypropylcellulose (e.g., low substituted Hydroxypropylcellulose), crosslinked polyvinylpyrrolidone, hydroxypropylcellulose, and starch (e.g., sodium glycolate starch, potato or tapioca starch).
  • hydroxypropylcellulose e.g., low substituted Hydroxypropylcellulose
  • starch e.g., sodium glycolate starch, potato or tapioca starch
  • Exemplary binding agents include starch (e.g., glycolate starch, cornstarch and starch paste), gelatin, sugars (e.g., sucrose, glucose, dextrose, dextrin, molasses, lactose, lactitol, mannitol, etc.), natural and synthetic gums (e.g., acacia, sodium alginate, extract of Irish moss, panwar gum, ghatti gum, mucilage of isapol husks, carboxymethylcellulose, methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose, microcrystalline cellulose, cellulose acetate, poly(vinyl-pyrrolidone), magnesium aluminum silicate (Veegum ® ), and larch arabogalactan), alginates, polyethylene oxide, polyethylene glycol, inorganic calcium salts, silicic acid, polymethacrylates, waxes
  • Exemplary preservatives include antioxidants, chelating agents, antimicrobial preservatives, antifungal preservatives, antiprotozoan preservatives, alcohol preservatives, acidic preservatives, and other preservatives.
  • the preservative is an antioxidant.
  • the preservative is a chelating agent.
  • antioxidants include alpha tocopherol, ascorbic acid, ascorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene, monothioglycerol, potassium metabisulfite, propionic acid, propyl gallate, sodium ascorbate, sodium bisulfite, sodium metabisulfite, and sodium sulfite.
  • Exemplary chelating agents include ethylenediaminetetraacetic acid (EDTA) and salts and hydrates thereof (e.g., sodium edetate, disodium edetate, trisodium edetate, calcium disodium edetate, dipotassium edetate, and the like), citric acid and salts and hydrates thereof (e.g., citric acid monohydrate), fumaric acid and salts and hydrates thereof, malic acid and salts and hydrates thereof, phosphoric acid and salts and hydrates thereof, and tartaric acid and salts and hydrates thereof.
  • EDTA ethylenediaminetetraacetic acid
  • salts and hydrates thereof e.g., sodium edetate, disodium edetate, trisodium edetate, calcium disodium edetate, dipotassium edetate, and the like
  • citric acid and salts and hydrates thereof e.g., citric acid mono
  • antimicrobial preservatives include benzalkonium chloride, benzethonium chloride, benzyl alcohol, bronopol, cetrimide, cetylpyridinium chloride, chlorhexidine, chlorobutanol,
  • Exemplary antifungal preservatives include butyl paraben, methyl paraben, ethyl paraben, propyl paraben, benzoic acid, hydroxybenzoic acid, potassium benzoate, potassium sorbate, sodium benzoate, sodium propionate, and sorbic acid.
  • Exemplary alcohol preservatives include ethanol, polyethylene glycol, phenol, phenolic compounds, bisphenol, chlorobutanol, hydroxybenzoate, and phenylethyl alcohol.
  • Exemplary acidic preservatives include vitamin A, vitamin C, vitamin E, beta-carotene, citric acid, acetic acid, dehydroacetic acid, ascorbic acid, sorbic acid, and phytic acid.
  • preservatives include tocopherol, tocopherol acetate, deteroxime mesylate, cetrimide, butylated hydroxyanisol (BHA), butylated hydroxytoluened (BHT), ethylenediamine, sodium lauryl sulfate (SLS), sodium lauryl ether sulfate (SLES), sodium bisulfite, sodium metabisulfite, potassium sulfite, potassium metabisulfite, Glydant ® Plus, Phenonip ® , methylparaben, Germall ® 115, Germaben ® II, Neolone ® , Kathon ® , and Euxyl ® .
  • Exemplary buffering agents include citrate buffer solutions, acetate buffer solutions, phosphate buffer solutions, ammonium chloride, calcium carbonate, calcium chloride, calcium citrate, calcium glubionate, calcium gluceptate, calcium gluconate, D-gluconic acid, calcium glycerophosphate, calcium lactate, propanoic acid, calcium levulinate, pentanoic acid, dibasic calcium phosphate, phosphoric acid, tribasic calcium phosphate, calcium hydroxide phosphate, potassium acetate, potassium chloride, potassium gluconate, potassium mixtures, dibasic potassium phosphate, monobasic potassium phosphate, potassium phosphate mixtures, sodium acetate, sodium bicarbonate, sodium chloride, sodium citrate, sodium lactate, dibasic sodium phosphate, monobasic sodium phosphate, sodium phosphate mixtures, tromethamine, magnesium hydroxide, aluminum hydroxide, alginic acid, pyrogen-free water, isotonic saline
  • Exemplary lubricating agents include agar, ethyl oleate, ethyl laurate, glycerin, blyceryl palmitostearate, magnesium oxide, magnesium stearate, mannitol, poloxamer, glycol, sodium stearyl, sorbitol, zinc stearate, magnesium stearate, calcium stearate, stearic acid, silica, talc, malt, glyceryl behanate, hydrogenated vegetable oils, polyethylene glycol, sodium benzoate, sodium acetate, sodium chloride, leucine, magnesium lauryl sulfate, sodium lauryl sulfate, and mixtures thereof.
  • Exemplary natural oils include almond, apricot kernel, avocado, babassu, bergamot, black current seed, borage, cade, camomile, canola, caraway, carnauba, castor, cinnamon, cocoa butter, coconut, cod liver, coffee, corn, cotton seed, emu, eucalyptus, evening primrose, fish, flaxseed, geraniol, gourd, grape seed, hazel nut, hyssop, isopropyl myristate, jojoba, kukui nut, lavandin, lavender, lemon, litsea cubeba, macademia nut, mallow, mango seed, meadowfoam seed, mink, nutmeg, olive, orange, orange roughy, palm, palm kernel, peach kernel, peanut, poppy seed, pumpkin seed, rapeseed, rice bran, rosemary, safflower, sandalwood, sasquana, savoury, sea
  • Exemplary synthetic oils include, but are not limited to, butyl stearate, caprylic triglyceride, capric triglyceride, cyclomethicone, diethyl sebacate, dimethicone 360, isopropyl myristate, mineral oil, octyldodecanol, oleyl alcohol, silicone oil, and mixtures thereof.
  • Liquid dosage forms for oral and parenteral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs.
  • the liquid dosage forms may comprise inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (e.g., cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof.
  • inert diluents commonly used in the art such as, for example, water or other solvents, so
  • the oral compositions can include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.
  • adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.
  • the conjugates described herein are mixed with solubilizing agents such as Cremophor ® , alcohols, oils, modified oils, glycols, polysorbates, cyclodextrins, polymers, and mixtures thereof.
  • solubilizing agents such as Cremophor ® , alcohols, oils, modified oils, glycols, polysorbates, cyclodextrins, polymers, and mixtures thereof.
  • injectable preparations for example, sterile injectable aqueous or oleaginous suspensions can be formulated according to the known art using suitable dispersing or wetting agents and suspending agents.
  • the sterile injectable preparation can be a sterile injectable solution, suspension, or emulsion in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol.
  • a nontoxic parenterally acceptable diluent or solvent for example, as a solution in 1,3-butanediol.
  • acceptable vehicles and solvents that can be employed are water, Ringer’s solution, U.S.P., and isotonic sodium chloride solution.
  • sterile, fixed oils are conventionally employed as a solvent or suspending medium.
  • any bland fixed oil can be employed including synthetic mono- or di-glycerides.
  • fatty acids such as oleic acid are used in the preparation of injectables.
  • injectable preparations of the compositions disclosed herein are in the form of a ready-to-use (“RTU”) preparation that can be directly administered to a subject.
  • the RTU preparation is a suspension.
  • the RTU preparation is a solution.
  • the RTU preparation is an emulsion.
  • injectable preparations of the compositions disclosed herein are in the form of a solid that is reconstituted prior to administration.
  • the solid is a lyophilized solid.
  • injectable preparations of the compositions disclosed herein are in the form of a liquid or suspension that is diluted prior to administration.
  • the injectable formulations can be sterilized, for example, by filtration through a bacterial- retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.
  • compositions for rectal or vaginal administration are typically suppositories which can be prepared by mixing the conjugates described herein with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol, or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active ingredient.
  • suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol, or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active ingredient.
  • Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules.
  • the active ingredient is mixed with at least one inert, pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate and/or (a) fillers or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic acid, (b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone, sucrose, and acacia, (c) humectants such as glycerol, (d) disintegrating agents such as agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, (e) solution retarding agents such as paraffin, (f) absorption accelerators such as quaternary ammonium compounds, (g) wetting agents such as, for example, cetyl alcohol and glycerol monostearate, (h) absorbents such as kaolin and bentonite clay, and (a) fillers or
  • the dosage form may include a buffering agent.
  • Solid compositions of a similar type can be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like.
  • the solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the art of pharmacology. They may optionally comprise opacifying agents and can be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner.
  • encapsulating compositions which can be used include polymeric substances and waxes.
  • Solid compositions of a similar type can be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polethylene glycols and the like.
  • the active ingredient can be in a micro-encapsulated form with one or more excipients as noted above.
  • the solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings, release controlling coatings, and other coatings well known in the pharmaceutical formulating art. In such solid dosage forms the active ingredient can be
  • Such dosage forms may comprise, as is normal practice, additional substances other than inert diluents, e.g., tableting lubricants and other tableting aids such a magnesium stearate and microcrystalline cellulose.
  • the dosage forms may comprise buffering agents. They may optionally comprise opacifying agents and can be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner.
  • Dosage forms for topical and/or transdermal administration of a compound described herein may include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants, and/or patches.
  • the active ingredient is admixed under sterile conditions with a pharmaceutically acceptable carrier or excipient and/or any needed preservatives and/or buffers as can be required.
  • the present disclosure contemplates the use of transdermal patches, which often have the added advantage of providing controlled delivery of an active ingredient to the body.
  • Such dosage forms can be prepared, for example, by dissolving and/or dispensing the active ingredient in the proper medium.
  • the rate can be controlled by either providing a rate controlling membrane and/or by dispersing the active ingredient in a polymer matrix and/or gel.
  • Suitable devices for use in delivering intradermal pharmaceutical compositions described herein include short needle devices. Intradermal compositions can be administered by devices which limit the effective penetration length of a needle into the skin. Alternatively or additionally, conventional syringes can be used in the classical mantoux method of intradermal administration. Jet injection devices which deliver liquid formulations to the dermis via a liquid jet injector and/or via a needle which pierces the stratum corneum and produces a jet which reaches the dermis are suitable.
  • Formulations suitable for topical administration include, but are not limited to, liquid and/or semi-liquid preparations such as liniments, lotions, oil-in-water and/or water-in-oil emulsions such as creams, ointments, and/or pastes, and/or solutions and/or suspensions.
  • Topically administrable formulations may, for example, comprise from about 1% to about 10% (w/w) active ingredient, although the concentration of the active ingredient can be as high as the solubility limit of the active ingredient in the solvent.
  • Formulations for topical administration may further comprise one or more of the additional ingredients described herein.
  • a pharmaceutical composition described herein can be prepared, packaged, and/or sold in a formulation suitable for pulmonary administration via the buccal cavity.
  • a formulation may comprise dry particles which comprise the active ingredient and which have a diameter in the range from about 0.5 to about 7 nanometers, or from about 1 to about 6 nanometers.
  • Such compositions are conveniently in the form of dry powders for administration using a device comprising a dry powder
  • U1197.70243WO00 114/193 #13587456v2 reservoir to which a stream of propellant can be directed to disperse the powder and/or using a self- propelling solvent/powder dispensing container such as a device comprising the active ingredient dissolved and/or suspended in a low-boiling propellant in a sealed container.
  • a self- propelling solvent/powder dispensing container such as a device comprising the active ingredient dissolved and/or suspended in a low-boiling propellant in a sealed container.
  • Such powders comprise particles wherein at least 98% of the particles by weight have a diameter greater than 0.5 nanometers and at least 95% of the particles by number have a diameter less than 7 nanometers. Alternatively, at least 95% of the particles by weight have a diameter greater than 1 nanometer and at least 90% of the particles by number have a diameter less than 6 nanometers.
  • Dry powder compositions may include a solid fine powder diluent such as sugar and are conveniently provided in a unit dose form.
  • Low boiling propellants generally include liquid propellants having a boiling point of below 65 °F at atmospheric pressure. Generally the propellant may constitute 50 to 99.9% (w/w) of the composition, and the active ingredient may constitute 0.1 to 20% (w/w) of the composition.
  • the propellant may further comprise additional ingredients such as a liquid non-ionic and/or solid anionic surfactant and/or a solid diluent (which may have a particle size of the same order as particles comprising the active ingredient).
  • compositions described herein formulated for pulmonary delivery may provide the active ingredient in the form of droplets of a solution and/or suspension.
  • Such formulations can be prepared, packaged, and/or sold as aqueous and/or dilute alcoholic solutions and/or suspensions, optionally sterile, comprising the active ingredient, and may conveniently be administered using any nebulization and/or atomization device.
  • Such formulations may further comprise one or more additional ingredients including, but not limited to, a flavoring agent such as saccharin sodium, a volatile oil, a buffering agent, a surface active agent, and/or a preservative such as methylhydroxybenzoate.
  • the droplets provided by this route of administration may have an average diameter in the range from about 0.1 to about 200 nanometers.
  • Formulations described herein as being useful for pulmonary delivery are useful for intranasal delivery of a pharmaceutical composition described herein.
  • Another formulation suitable for intranasal administration is a coarse powder comprising the active ingredient and having an average particle from about 0.2 to 500 micrometers. Such a formulation is administered by rapid inhalation through the nasal passage from a container of the powder held close to the nares.
  • Formulations for nasal administration may, for example, comprise from about as little as 0.1% (w/w) to as much as 100% (w/w) of the active ingredient, and may comprise one or more of the additional ingredients described herein.
  • a pharmaceutical composition described herein can be prepared, packaged, and/or sold in a formulation for buccal administration.
  • formulations may, for example, be in the form of tablets and/or lozenges made using conventional methods, and may contain, for example, 0.1 to 20% (w/w) active ingredient, the balance comprising an orally dissolvable and/or degradable composition and, optionally, one or more of the additional ingredients described herein.
  • formulations for buccal administration may comprise a powder and/or an aerosolized and/or atomized solution and/or suspension comprising the active ingredient.
  • U1197.70243WO00 115/193 #13587456v2 powdered, aerosolized, and/or aerosolized formulations when dispersed, may have an average particle and/or droplet size in the range from about 0.1 to about 200 nanometers, and may further comprise one or more of the additional ingredients described herein.
  • a pharmaceutical composition described herein can be prepared, packaged, and/or sold in a formulation for ophthalmic administration. Such formulations may, for example, be in the form of eye drops including, for example, a 0.1-1.0% (w/w) solution and/or suspension of the active ingredient in an aqueous or oily liquid carrier or excipient.
  • Such drops may further comprise buffering agents, salts, and/or one or more other of the additional ingredients described herein.
  • Other opthalmically- administrable formulations which are useful include those which comprise the active ingredient in microcrystalline form and/or in a liposomal preparation. Ear drops and/or eye drops are also contemplated as being within the scope of this disclosure.
  • compositions suitable for administration to humans in order to render the compositions suitable for administration to various animals is well understood, and the ordinarily skilled veterinary pharmacologist can design and/or perform such modification with ordinary experimentation.
  • Compounds provided herein are typically formulated in dosage unit form for ease of administration and uniformity of dosage. It will be understood, however, that the total daily usage of the compositions described herein will be decided by a physician within the scope of sound medical judgment.
  • the specific therapeutically effective dose level for any particular subject or organism will depend upon a variety of factors including the disease being treated and the severity of the disorder; the activity of the specific active ingredient employed; the specific composition employed; the age, body weight, general health, sex, and diet of the subject; the time of administration, route of administration, and rate of excretion of the specific active ingredient employed; the duration of the treatment; drugs used in combination or coincidental with the specific active ingredient employed; and like factors well known in the medical arts.
  • the compounds and compositions provided herein can be administered by any route, including enteral (e.g., oral), parenteral, intravenous, intramuscular, intra-arterial, intramedullary, intrathecal, subcutaneous, intraventricular, transdermal, interdermal, rectal, intravaginal, intraperitoneal, topical (as by powders, ointments, creams, and/or drops), mucosal, nasal, bucal, sublingual; by intratracheal instillation, bronchial instillation, and/or inhalation; and/or as an oral spray, nasal spray, and/or aerosol.
  • enteral e.g., oral
  • parenteral intravenous, intramuscular, intra-arterial, intramedullary
  • intrathecal subcutaneous, intraventricular, transdermal, interdermal, rectal, intravaginal, intraperitoneal
  • topical as by powders, ointments, creams, and/or drops
  • mucosal nasal,
  • Specifically contemplated routes are oral administration, intravenous administration (e.g., systemic intravenous injection), regional administration via blood and/or lymph supply, and/or direct administration to an affected site.
  • intravenous administration e.g., systemic intravenous injection
  • regional administration via blood and/or lymph supply e.g., systemic intravenous injection
  • direct administration e.g., direct administration to an affected site.
  • the most appropriate route of administration will depend upon a variety of factors including the nature of the agent (e.g., its
  • the compound or pharmaceutical composition described herein is suitable for topical administration to the eye of a subject.
  • the exact amount of a compound required to achieve an effective amount will vary from subject to subject, depending, for example, on species, age, and general condition of a subject, severity of the side effects or disorder, identity of the particular compound, mode of administration, and the like.
  • An effective amount may be included in a single dose (e.g., single oral dose) or multiple doses (e.g., multiple oral doses).
  • any two doses of the multiple doses include different or substantially the same amounts of a compound described herein.
  • the frequency of administering the multiple doses to the subject or applying the multiple doses to the tissue or cell is three doses a day, two doses a day, one dose a day, one dose every other day, one dose every third day, one dose every week, one dose every two weeks, one dose every three weeks, or one dose every four weeks.
  • the frequency of administering the multiple doses to the subject or applying the multiple doses to the tissue or cell is one dose per day.
  • the frequency of administering the multiple doses to the subject or applying the multiple doses to the tissue or cell is two doses per day. In certain embodiments, the frequency of administering the multiple doses to the subject or applying the multiple doses to the tissue or cell is three doses per day. In certain embodiments, when multiple doses are administered to a subject or applied to a tissue or cell, the duration between the first dose and last dose of the multiple doses is one day, two days, four days, one week, two weeks, three weeks, one month, two months, three months, four months, six months, nine months, one year, two years, three years, four years, five years, seven years, ten years, fifteen years, twenty years, or the lifetime of the subject, tissue, or cell.
  • a dose (e.g., a single dose, or any dose of multiple doses) described herein includes independently between 0.1 ⁇ g and 1 ⁇ g, between 0.001 mg and 0.01 mg, between 0.01 mg and 0.1 mg, between 0.1 mg and 1 mg, between 1 mg and 3 mg, between 3 mg and 10 mg, between 10 mg and 30 mg, between 30 mg and 100 mg, between 100 mg and 300 mg, between 300 mg and 1,000 mg, or between 1 g and 10 g, inclusive, of a compound described herein.
  • a dose described herein includes independently between 1 mg and 3 mg, inclusive, of a compound described herein. In certain embodiments, a dose described herein includes independently between 3 mg and 10 mg, inclusive, of a compound described herein. In certain embodiments, a dose described herein includes independently between 10
  • a dose described herein includes independently between 30 mg and 100 mg, inclusive, of a compound described herein.
  • Dose ranges as described herein provide guidance for the administration of provided pharmaceutical compositions to an adult.
  • the amount to be administered to, for example, a child or an adolescent can be determined by a medical practitioner or person skilled in the art and can be lower or the same as that administered to an adult.
  • a compound or composition, as described herein, can be administered in combination with one or more additional pharmaceutical agents (e.g., therapeutically and/or prophylactically active agents).
  • the compounds or compositions can be administered in combination with additional pharmaceutical agents that improve their activity (e.g., activity (e.g., potency and/or efficacy) in treating a disease or disorder in a subject in need thereof, in preventing a disease or disorder in a subject in need thereof, in reducing the risk to develop a disease or disorder in a subject in need thereof, and/or in cellular engineering in a subject or cell), improve bioavailability, improve safety, reduce drug resistance, reduce and/or modify metabolism, inhibit excretion, and/or modify distribution in a subject or cell.
  • the therapy employed may achieve a desired effect for the same disorder, and/or it may achieve different effects.
  • a pharmaceutical composition described herein including a compound described herein and an additional pharmaceutical agent shows a synergistic effect that is absent in a pharmaceutical composition including one of the compound and the additional pharmaceutical agent, but not both.
  • the additional pharmaceutical agent achieves a desired effect for the same disorder.
  • the additional pharmaceutical agent achieves different effects.
  • the compound or composition can be administered concurrently with, prior to, or subsequent to one or more additional pharmaceutical agents, which may be useful as, e.g., combination therapies.
  • Pharmaceutical agents include therapeutically active agents.
  • Pharmaceutical agents also include prophylactically active agents.
  • Pharmaceutical agents include small organic molecules such as drug compounds (e.g., compounds approved for human or veterinary use by the U.S.
  • CFR Code of Federal Regulations
  • proteins proteins, carbohydrates, monosaccharides, oligosaccharides, polysaccharides, nucleoproteins, mucoproteins, lipoproteins, synthetic polypeptides or proteins, small molecules linked to proteins, glycoproteins, steroids, nucleic acids, DNAs, RNAs, nucleotides, nucleosides, oligonucleotides, antisense oligonucleotides, lipids, hormones, vitamins, and cells.
  • CFR Code of Federal Regulations
  • the additional pharmaceutical agent is a pharmaceutical agent useful for treating and/or preventing a disease or disorder (e.g., proliferative disease, hematological disease, neurological disease, painful condition, psychiatric disorder, or metabolic disorder).
  • a disease or disorder e.g., proliferative disease, hematological disease, neurological disease, painful condition, psychiatric disorder, or metabolic disorder.
  • Each additional pharmaceutical agent may be administered at a dose and/or on a time schedule determined for that pharmaceutical agent.
  • the additional pharmaceutical agents may also be administered together with each other and/or with the compound or composition described herein in a single dose or composition or administered separately
  • the additional pharmaceutical agents include, but are not limited to, anti-proliferative agents, anti-cancer agents, anti-angiogenesis agents, steroidal or non-steroidal anti-inflammatory agents, immunosuppressants, anti-bacterial agents, anti-viral agents, cardiovascular agents, cholesterol- lowering agents, anti-diabetic agents, anti-allergic agents, contraceptive agents, pain-relieving agents, anesthetics, anti–coagulants, inhibitors of an enzyme, steroidal agents, steroidal or antihistamine, antigens, vaccines, antibodies, decongestant, sedatives, opioids, analgesics, anti–pyretics, hormones, and prostaglandins.
  • the additional pharmaceutical agent is an anti- proliferative agent. In certain embodiments, the additional pharmaceutical agent is an anti-cancer agent. In certain embodiments, the additional pharmaceutical agent is an anti-viral agent. In certain embodiments, the additional pharmaceutical agent is an binder or inhibitor of a protein kinase.
  • the additional pharmaceutical agent is selected from the group consisting of epigenetic or transcriptional modulators (e.g., DNA methyltransferase inhibitors, histone deacetylase inhibitors (HDAC inhibitors), lysine methyltransferase inhibitors), antimitotic drugs (e.g., taxanes and vinca alkaloids), hormone receptor modulators (e.g., estrogen receptor modulators and androgen receptor modulators), cell signaling pathway inhibitors (e.g., tyrosine protein kinase inhibitors), modulators of protein stability (e.g., proteasome inhibitors), Hsp90 inhibitors, glucocorticoids, all- trans retinoic acids, and other agents that promote differentiation.
  • epigenetic or transcriptional modulators e.g., DNA methyltransferase inhibitors, histone deacetylase inhibitors (HDAC inhibitors), lysine methyltransferase inhibitors
  • antimitotic drugs e.g., taxanes and vinca
  • the compounds described herein or pharmaceutical compositions can be administered in combination with an anti-cancer therapy including, but not limited to, surgery, radiation therapy, transplantation (e.g., stem cell transplantation, bone marrow transplantation), immunotherapy, and chemotherapy.
  • an anti-cancer therapy including, but not limited to, surgery, radiation therapy, transplantation (e.g., stem cell transplantation, bone marrow transplantation), immunotherapy, and chemotherapy.
  • Additional pharmaceutical agents include small organic molecules such as drug compounds (e.g., compounds approved by the US Food and Drug Administration as provided in the Code of Federal Regulations (CFR)), peptides, proteins, carbohydrates, monosaccharides, oligosaccharides, polysaccharides, nucleoproteins, mucoproteins, lipoproteins, synthetic polypeptides or proteins, small molecules linked to proteins, glycoproteins, steroids, nucleic acids, DNAs, RNAs, nucleotides, nucleosides, oligonucleotides, antisense oligonucleotides, lipids, hormones, vitamins and cells.
  • kits e.g., pharmaceutical packs.
  • kits provided may comprise a pharmaceutical composition or compound described herein and a container (e.g., a vial, ampule, bottle, syringe, and/or dispenser package, or other suitable container).
  • a container e.g., a vial, ampule, bottle, syringe, and/or dispenser package, or other suitable container.
  • provided kits may optionally further include a second container comprising a pharmaceutical excipient for dilution or suspension of a pharmaceutical composition or compound
  • kits including a first container comprising a compound or pharmaceutical composition described herein.
  • the kits are useful for treating a disease or disorder (e.g., proliferative disease, hematological disease, neurological disease, painful condition, psychiatric disorder, or metabolic disorder) in a subject in need thereof.
  • a disease or disorder e.g., proliferative disease, hematological disease, neurological disease, painful condition, psychiatric disorder, or metabolic disorder
  • kits are useful for preventing a disease or disorder (e.g., proliferative disease, hematological disease, neurological disease, painful condition, psychiatric disorder, or metabolic disorder) in a subject in need thereof.
  • the kits are useful for reducing the risk of developing a disease or disorder (e.g., proliferative disease, hematological disease, neurological disease, painful condition, psychiatric disorder, or metabolic disorder) in a subject in need thereof.
  • the kits are useful for cellular engineering in a subject or cell.
  • a kit described herein further includes instructions for using the kit.
  • a kit described herein may also include information as required by a regulatory agency such as the U.S.
  • kits and instructions provide for treating a disease or disorder (e.g., proliferative disease, hematological disease, neurological disease, painful condition, psychiatric disorder, or metabolic disorder) in a subject in need thereof.
  • a disease or disorder e.g., proliferative disease, hematological disease, neurological disease, painful condition, psychiatric disorder, or metabolic disorder
  • the kits and instructions provide for preventing a disease or disorder (e.g., proliferative disease, hematological disease, neurological disease, painful condition, psychiatric disorder, or metabolic disorder) in a subject in need thereof.
  • kits and instructions provide for reducing the risk of developing a disease or disorder (e.g., proliferative disease, hematological disease, neurological disease, painful condition, psychiatric disorder, or metabolic disorder) in a subject in need thereof.
  • a disease or disorder e.g., proliferative disease, hematological disease, neurological disease, painful condition, psychiatric disorder, or metabolic disorder
  • the kits and instructions provide for cellular engineering in a subject or cell.
  • a kit described herein may include one or more additional pharmaceutical agents described herein as a separate composition.
  • a provided composition (a composition described herein, a composition of the present disclosure) is a composition comprising a compound of any of the formulae herein (e.g., Formulae (I), (I′), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), (XI)), or a pharmaceutically acceptable salt thereof.
  • a provided composition is a composition comprising a compound of any of the formulae herein (e.g., Formulae (I), (I′), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), (XI)), or a salt thereof.
  • a provided composition is a composition comprising a compound of any of the formulae herein (e.g., Formulae (I), (I′), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), (XI)).
  • the present disclosure provides a method of treating or preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a provided compound or provided composition.
  • the present disclosure provides a method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a provided compound or provided composition.
  • the present disclosure provides a method of preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a provided compound or provided composition.
  • the present disclosure provides a method of treating or preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formulae (I), (I′), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), or (XI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the present disclosure provides a method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formulae (I), (I′), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), or (XI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the present disclosure provides a method of preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formulae (I), (I′), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), or (XI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the present disclosure provides a method of treating or preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the present disclosure provides a method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the present disclosure provides a method of preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the present disclosure provides a method of treating or preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (I′), or a pharmaceutically
  • the present disclosure provides a method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (I′), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the present disclosure provides a method of preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (I′), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the present disclosure provides a method of treating or preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (II), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the present disclosure provides a method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (II), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the present disclosure provides a method of preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (II), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the present disclosure provides a method of treating or preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (III), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the present disclosure provides a method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (III), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the present disclosure provides a method of preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (III), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the present disclosure provides a method of treating or preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (IV), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the present disclosure provides a method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (IV), or a pharmaceutically acceptable salt thereof, or a pharmaceutical
  • the present disclosure provides a method of preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (IV), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the present disclosure provides a method of treating or preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (V), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the present disclosure provides a method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (V), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the present disclosure provides a method of preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (V), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the present disclosure provides a method of treating or preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (VI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the present disclosure provides a method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (VI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the present disclosure provides a method of preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (VI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the present disclosure provides a method of treating or preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (VII), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the present disclosure provides a method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (VII), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the present disclosure provides a method of preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (VII), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the present disclosure provides a method of treating or preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (VIII), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the present disclosure provides a method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (VIII), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the present disclosure provides a method of preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (VIII), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the present disclosure provides a method of treating or preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (IX), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the present disclosure provides a method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (IX), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the present disclosure provides a method of preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (IX), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the present disclosure provides a method of treating or preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (X), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the present disclosure provides a method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (X), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the present disclosure provides a method of preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (X), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the present disclosure provides a method of treating or preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (XI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the present disclosure provides a method of treating or preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (XI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • U1197.70243WO00 124/193 #13587456v2 disclosure provides a method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (XI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the present disclosure provides a method of preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (XI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the present disclosure provides a provided compound or provided composition for use in the treatment or prevention of a disease or disorder.
  • the present disclosure provides a provided compound or provided composition for use in the treatment of a disease or disorder. In some embodiments, the present disclosure provides a provided compound or provided composition for use in the prevention of a disease or disorder. [0418] In another aspect, the present disclosure provides a compound of Formulae (I), (I′), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), or (XI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, for use in the treatment or prevention of a disease or disorder.
  • the present disclosure provides a compound of Formulae (I), (I′), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), or (XI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, for use in the treatment of a disease or disorder.
  • the present disclosure provides a compound of Formulae (I), (I′), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), or (XI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, for use in the prevention of a disease or disorder.
  • the present disclosure provides a provided compound or provided composition for use in the manufacture of a medicament for the treatment or prevention of a disease or disorder. In some embodiments, the present disclosure provides a provided compound or provided composition for use in the manufacture of a medicament for the treatment of a disease or disorder. In some embodiments, the present disclosure provides a provided compound or provided composition for use in the manufacture of a medicament for the prevention of a disease or disorder.
  • the present disclosure provides a compound of Formulae (I), (I′), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), or (XI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, for use in the manufacture of a medicament for the treatment or prevention of a disease or disorder.
  • the present disclosure provides a compound of Formulae (I), (I′), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), or (XI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, for use in the manufacture of a medicament for the treatment of a disease or disorder.
  • the present disclosure provides a compound of Formulae (I), (I′), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), or (XI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, for use in the manufacture of a medicament for the prevention of a disease or disorder.
  • the method of treating or preventing a disease or disorder comprises modulating a target protein.
  • modulating the target protein comprises inhibition of the target protein.
  • modulating the target protein comprises agonism of the target protein.
  • modulating the target protein comprises antagonism of the target protein.
  • the disease or disorder is associated with the target protein.
  • the disease or disorder is associated with signal transducer and activator of transcription 3 (STAT3), Toll-like receptor 7 (TLR7), indoleamine 2,3-dioxygenase (IDO), nitric oxide synthase (NOS), arginase, programmed death-ligand 1 (PD-L1), protein kinase B, cluster of differentiation 73 (CD73), or stimulator of interferon genes (STING).
  • STAT3 signal transducer and activator of transcription 3
  • TLR7 Toll-like receptor 7
  • the disease or disorder is associated with indoleamine 2,3-dioxygenase (IDO). In some embodiments, the disease or disorder is associated with nitric oxide synthase (NOS). In some embodiments, the disease or disorder is associated with arginase. In some embodiments, the disease or disorder is associated with programmed death-ligand 1 (PD-L1). In some embodiments, the disease or disorder is associated with protein kinase B. In some embodiments, the disease or disorder is associated with cluster of differentiation 73 (CD73). In some embodiments, the disease or disorder is associated with stimulator of interferon genes (STING).
  • IDO indoleamine 2,3-dioxygenase
  • NOS nitric oxide synthase
  • the disease or disorder is associated with arginase.
  • the disease or disorder is associated with programmed death-ligand 1 (PD-L1).
  • the disease or disorder is associated with protein kinase B.
  • the disease or disorder is associated with cluster of differentiation 73
  • the disease or disorder is a genetic disease, proliferative disease, hematological disease, neurological disease, liver disease, spleen disease, lung disease, painful condition, psychiatric disorder, musculoskeletal disease, a metabolic disorder, inflammatory disease, autoimmune disease, or microbial infection.
  • the disease or disorder is a proliferative disease.
  • the proliferative disease is cancer (e.g., a solid tumor (e.g., brain tumor (e.g., glioma, glioblastoma), pancreatic cancer, ovarian cancer)).
  • the proliferative disease comprises a solid tumor (e.g., brain tumor (e.g., glioma, glioblastoma), pancreatic cancer, ovarian cancer).
  • the proliferative disease is a solid tumor (e.g., brain tumor (e.g., glioma, glioblastoma), pancreatic cancer, ovarian cancer).
  • the proliferative disease is a brain tumor (e.g., glioma, glioblastoma), pancreatic cancer, or ovarian cancer.
  • the proliferative disease is a brain tumor (e.g., glioma, glioblastoma).
  • the proliferative disease is glioma. In some embodiments, the proliferative disease is glioblastoma. In some embodiments, the proliferative disease is pancreatic cancer. In some embodiments, the proliferative disease is ovarian cancer. [0423] In some embodiments, the disease or disorder is a liver disease or a spleen disease. In some embodiments, the disease or disorder is a liver disease. In some embodiments, the disease or disorder is a spleen disease.
  • the disease or disorder is a microbial infection (e.g., sepsis). In some embodiments, the disease or disorder is sepsis.
  • the present disclosure provides a method of modulating a target protein, the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a provided compound or a provided composition.
  • the present disclosure provides a method of modulating a target protein, the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formulae (I), (I′), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), or (XI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • modulating the target protein comprises inhibition of the target protein.
  • modulating the target protein comprises agonism of the target protein.
  • modulating the target protein comprises antagonism of the target protein.
  • the target protein is signal transducer and activator of transcription 3 (STAT3), Toll-like receptor 7 (TLR7), indoleamine 2,3-dioxygenase (IDO), nitric oxide synthase (NOS), arginase, programmed death-ligand 1 (PD-L1), protein kinase B, cluster of differentiation 73 (CD73), or stimulator of interferon genes (STING).
  • STAT3 signal transducer and activator of transcription 3
  • the target protein is Toll- like receptor 7 (TLR7).
  • the target protein is indoleamine 2,3-dioxygenase (IDO).
  • the target protein is nitric oxide synthase (NOS). In some embodiments, the target protein is arginase. In some embodiments, the target protein is programmed death-ligand 1 (PD-L1). In some embodiments, the target protein is protein kinase B. In some embodiments, the target protein is cluster of differentiation 73 (CD73). In some embodiments, the target protein is stimulator of interferon genes (STING). [0429] In some embodiments, the present disclosure provides a method of modulating a target protein, the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • NOS nitric oxide synthase
  • the target protein is arginase.
  • the target protein is programmed death-ligand 1 (PD-L1).
  • the target protein protein kinase B.
  • the target protein is cluster of differentiation 73 (CD73
  • the method comprises inhibition of STAT3, agonism of TLR7, inhibition of IDO, inhibition of NOS, inhibition of arginase, inhibition of PD-L1, inhibition of protein kinase B, inhibition of CD73, or agonism of STING.
  • the method comprises inhibition of STAT3.
  • the method comprises agonism of TLR7.
  • the method comprises inhibition of IDO.
  • the method comprises inhibition of NOS.
  • the method comprises inhibition of arginase.
  • the method comprises inhibition of PD-L1.
  • the method comprises inhibition of protein kinase B.
  • the method comprises inhibition of CD73.
  • the method comprises agonism of STING.
  • the present disclosure provides a method of modulating a target protein, the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (I′), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the method comprises inhibition of STAT3, agonism of TLR7, inhibition of IDO, inhibition of NOS, inhibition of arginase, inhibition of PD-L1, inhibition of protein kinase B, inhibition of CD73, or agonism of STING.
  • the method comprises inhibition of STAT3.
  • the method comprises agonism of TLR7.
  • the method comprises inhibition of IDO. In some embodiments, the method comprises inhibition of NOS. In some embodiments, the method comprises inhibition of arginase. In some embodiments, the method comprises inhibition of PD-L1. In some embodiments, the method comprises inhibition of protein kinase B. In some embodiments, the method comprises inhibition of CD73. In some embodiments, the method comprises agonism of STING. [0431] In some embodiments, the present disclosure provides a method of modulating a target protein, the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (II), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the target protein is STAT3.
  • the method comprises inhibition of STAT3.
  • the present disclosure provides a method of modulating a target protein, the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (III), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the target protein is TLR7.
  • the method comprises agonism of TLR7.
  • the present disclosure provides a method of modulating a target protein, the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (IV), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the target protein is IDO.
  • the method comprises inhibition of IDO.
  • the present disclosure provides a method of modulating a target protein, the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (V), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the target protein is NOS. In some embodiments, the method comprises inhibition of NOS. [0435] In some embodiments, the present disclosure provides a method of modulating a target protein, the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (VI), or a pharmaceutically
  • the target protein is arginase. In some embodiments, the method comprises inhibition of arginase.
  • the present disclosure provides a method of modulating a target protein, the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (VII), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the target protein is PD-L1. In some embodiments, the method comprises inhibition of PD-L1.
  • the present disclosure provides a method of modulating a target protein, the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (VIII), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the target protein is protein kinase B.
  • the method comprises inhibition of protein kinase B.
  • the present disclosure provides a method of modulating a target protein, the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (IX), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the target protein is CD73.
  • the method comprises inhibition of CD73.
  • the present disclosure provides a method of modulating a target protein, the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (X), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the target protein is STING. In some embodiments, the method comprises agonism of STING. [0440] In some embodiments, the present disclosure provides a method of modulating a target protein, the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (XI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the target protein is STAT3. In some embodiments, the method comprises inhibition of STAT3.
  • the present disclosure provides a method of modulating signal transducer and activator of transcription 3 (STAT3), the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (II), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the modulating STAT3 comprises inhibition of STAT3.
  • the present disclosure provides a method of modulating Toll-like receptor 7 (TLR7), the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (III), or a pharmaceutically
  • the modulating TLR7 comprises agonism of TLR7.
  • the present disclosure provides a method of modulating indoleamine 2,3- dioxygenase (IDO), the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (IV), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the modulating IDO comprises inhibition of IDO.
  • the present disclosure provides a method of modulating nitric oxide synthase (NOS), the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (V), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • NOS nitric oxide synthase
  • the modulating NOS comprises inhibition of NOS.
  • the present disclosure provides a method of modulating arginase, the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (VI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the modulating arginase comprises inhibition of arginase.
  • the present disclosure provides a method of modulating programmed death- ligand 1 (PD-L1), the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (VII), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the modulating PD-L1 comprises inhibition of PD-L1.
  • the present disclosure provides a method of modulating protein kinase B, the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (VIII), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the modulating protein kinase B comprises inhibition of protein kinase B.
  • the present disclosure provides a method of modulating cluster of differentiation 73 (CD73), the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (IX), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the modulating CD73 comprises inhibition of CD73.
  • the present disclosure provides a method of modulating stimulator of interferon genes (STING), the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (X), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the modulating STING comprises agonism of STING.
  • the present disclosure provides a method of modulating signal transducer and activator of transcription 3 (STAT3), the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (XI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the modulating STAT3 comprises inhibition of STAT3.
  • the administration is by injection.
  • the subject is an animal. The animal may be of either sex and may be at any stage of development. In some embodiments, the subject described herein is a human.
  • the subject is a non-human animal. In some embodiments, the subject is a mammal. In some embodiments, the subject is a non-human mammal. In some embodiments, the subject is a domesticated animal, such as a dog, cat, cow, pig, horse, sheep, or goat. In some embodiments, the subject is a companion animal, such as a dog or cat. In some embodiments, the subject is a livestock animal, such as a cow, pig, horse, sheep, or goat. In some embodiments, the subject is a zoo animal. In another embodiment, the subject is a research animal, such as a rodent (e.g., mouse, rat), dog, pig, or non-human primate.
  • a rodent e.g., mouse, rat
  • the animal is a genetically engineered animal.
  • the animal is a transgenic animal (e.g., transgenic mice and transgenic pigs).
  • the subject is a fish or reptile.
  • Z n is an nth generation polyamidoamine (PAMAM) dendrimer; n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; each instance of R A is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; each of p and p1 is independently an integer between 1 and 2 (n+2) , inclusive; each of q and q1 is independently an integer between 0 and 2 (n+2) -1, inclusive; the sum of p and q is 2 (n+2)
  • PAMAM polyamidoamine
  • the present disclosure provides a method of preparing a compound of Formula (II-d): or a pharmaceutically acceptable salt thereof, the method comprising: providing a reaction vessel; adding to the reaction vessel a compound of Formula (XIII-b): or a salt thereof;
  • Z n is an nth generation polyamidoamine (PAMAM) dendrimer; n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; each instance of R A is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; each of p and p2 is independently an integer between 1 and 2 (n+2) , inclusive; each of q and q2 is independently an integer between 0 and 2 (n+2) -1,
  • the present disclosure provides a method of preparing a compound of Formula (XI-d): or a pharmaceutically acceptable salt thereof, the method comprising: providing a reaction vessel; adding to the reaction vessel a compound of Formula (XIII-b):
  • Z n is an nth generation polyamidoamine (PAMAM) dendrimer; n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; each instance of R A is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; each of p and p2 is independently an integer between 1 and 2 (n+2) , inclusive; each of q and q2 is independently an integer between 0 and 2 (PAMAM) dendrimer; n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; each instance of R A is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl,
  • the present disclosure provides a compound, or a pharmaceutically acceptable salt thereof, made by the method comprising: providing a reaction vessel; adding to the reaction vessel a compound of Formula (XIII-a): (XIII-a), or a salt thereof; adding to the reaction vessel a compound of Formula (XIV-a):
  • Z n is an nth generation polyamidoamine (PAMAM) dendrimer
  • n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10
  • each instance of R A is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl
  • p1 is an integer between 1 and 2 (n+2) , inclusive
  • q1 is an integer between 0 and 2 (n+2) -1, inclusive
  • the sum of p1 and q1 is 2 (n+2) .
  • the method is according to conditions provided in the Examples.
  • the present disclosure provides a compound, or a pharmaceutically acceptable salt thereof, made by the method comprising: providing a reaction vessel; adding to the reaction vessel a compound of Formula (XIII-b): or a salt thereof; adding to the reaction vessel a compound of Formula (XIV-b): or a salt thereof; and obtaining the compound, or pharmaceutically acceptable salt thereof; wherein: Z n is an nth generation polyamidoamine (PAMAM) dendrimer; n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10;
  • PAMAM polyamidoamine
  • each instance of R A is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl;
  • p2 is an integer between 1 and 2 (n+2) , inclusive;
  • q2 is an integer between 0 and 2 (n+2) -1, inclusive; and the sum of p2 and q2 is 2 (n+2) .
  • the method is according to conditions provided in the Examples.
  • the present disclosure provides a compound, or a pharmaceutically acceptable salt thereof, made by the method comprising: providing a reaction vessel; adding to the reaction vessel a compound of Formula (XIII-b): or a salt thereof; adding to the reaction vessel a compound of Formula (XIV-c): or a salt thereof; and obtaining the compound, or pharmaceutically acceptable salt thereof; wherein: Z n is an nth generation polyamidoamine (PAMAM) dendrimer; n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; each instance of R A is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optional
  • the NH2 surface dendrimer was used as such after the evaporation of methanol from the stock solution.
  • the OH and SA surface dendrimers were further functionalized with amine terminals to conjugate Cy5 mono NHS ester (Cytiva).
  • Cy5-labeling of OH dendrimers Step 1: Fmoc-GABA-OH (Sigma- Aldrich) was coupled with G6-OH using PyBOP (Merck) as a coupling reagent to produce an intermediate with protecting group Fmoc.
  • Step 2 The Fmoc protecting group was removed using piperidine (Sigma-Aldrich) – DMF (Sigma-Aldrich) mixture to produce bi-functional dendrimers.
  • Step 3 The conjugation of Cy5 mono NHS ester was carried out in the presence of borate buffer (pH 8.5) with pure bifunctional dendrimer to produce G6-OH-Cy5 conjugate. Similarly, amine surface G6 dendrimer was labeled with Cy5 using borate buffer (pH 8.5).
  • step 1 For the Cy5-labeling of SA dendrimers, in step 1, G6 succinamic acid surface dendrimer was coupled with N-Fmoc 1,5-diaminobutane hydrobromide (Sigma-Aldrich) using EDC.HCl (Sigma-Aldrich) as a coupling reagent. In step 2, the successful deprotection of Fmoc using piperidine (Sigma-Aldrich) DMF (Sigma-Aldrich) mixture resulted in bi-functional dendrimers.
  • piperidine Sigma-Aldrich
  • DMF Sigma-Aldrich
  • the GL261 glioma cells were cultured in Dulbecco’s Modified Eagles Medium (DMEM) (Invitrogen) with 1% penicillin-streptomycin (Invitrogen) and 10% fetal bovine serum (FBS) (Thermo Scientific). Cells were expanded in T75 flasks (Falcon) in a humidified incubator (Thermo Scientific) at 37°C with 5% CO 2 . All cell lines tested negative for mycoplasma based on DNA-based PCR tests. [0465] M-MDSC induction and culture.
  • DMEM Modified Eagles Medium
  • FBS fetal bovine serum
  • the dendrimer solutions were filtered through a 0.22 ⁇ m 13mm polyether sulfone (PES) syringe filter (Cytiva) before the size measurement within a 1 mL cuvette (Sarstedt).
  • PES polyether sulfone
  • To measure the ⁇ -potential, dendrimers were diluted in 1 ⁇ PBS buffer (pH 7.4) at 0.3mg/mL.
  • the dendrimer solutions were also filtered before the ⁇ -potential measurement within an omega cuvette (Anton Paar).
  • dendrimers were incubated with mouse serum from C57BL/6j mice (in-house generated) for 30min at 37°C at a concentration of 0.86 mg/mL to allow the formation of the dendrimer-serum protein complex.
  • mice and in vivo tumor models Transgenic CCR2 RFP/WT /CX3CR1 GFP/WT C57BL/6 mice were bred in-house at the University of Florida animal facility.
  • CCR2 RFP/WT /CX3CR1 GFP/WT were generated by cross-breeding CCR2 deficient mice (CCR2 RFP/RFP [B6.129(Cg)-CCR2 tm2.1lfv /J]) and CX3CR1 deficient mice (CX3CR1 GFP/GFP [B6.129P-CX3CR1 tm1Litt /J]). Wildtype C57BL/6 mice were purchased from The Jackson Laboratory. All procedures involving animal housing, care, and surgical procedures were following the guidelines of the University of Florida Institutional Animal Care and Use Committee. [0468] Mouse models of Glioma.
  • mice were anesthetized by controlled isoflurane inhalation, and their heads were shaved before intravenous analgesia administration.
  • Surgical sites were prepared using 2-3mm incisions at the midline of the skull.
  • Stereotactic injection of 2 ⁇ L at 1 ⁇ L/min (5.0 x 10 4 ) cells suspended in methylcellulose was performed at 2mm lateral from the bregma using a Hamilton syringe autonomously controlled by a micro-fluidic injection apparatus (Stoelting).
  • Post-injection the dermal incision was closed via suture and bone wax application. Animals were placed on a cage warmer for post-surgical monitoring.
  • tumor tissue was minced using a regular single-edge razor blade until a viscous suspension of cells was generated.
  • Cells were then transferred to a 50mL conical (Falcon) filled with Accumax dissociation buffer (Innovative Cell Technologies) and incubated in a 37°C water bath for 5 minutes.
  • Suspensions were then oscillated through a 1mL single-channel pipet tip for 40 cycles and strained through a 40 ⁇ m strainer into a 50mL conical, followed by dilution with 5mL FACS buffer (10% FBS, 1 ⁇ HBSS).
  • the resulting tumor cell interface between Percoll layers was removed (1mL) by a single channel pipet and transferred to a 1.5mL microcentrifuge tube.
  • Cells were centrifuged at 500 ⁇ g for 5 minutes at 4°C and washed and resuspended with ice-cold FACS Buffer.
  • Femurs were harvested and ends clipped with dissecting scissors after connective tissues were removed.
  • the isolated femurs were placed in 0.5mL microcentrifuge tubes with an 18G needle pierced bottom, cap removed, and tube nested within a secondary 1.5 microcentrifuge tube containing 100 ⁇ L ice cold ACK lysis buffer (Gibco).
  • Microcentrifuge tubes with femurs were centrifuged at 5,700 RPM for 20 seconds at 21°C to capture bone marrow.
  • Spleens were excised and transferred to a petri dish on ice for mincing using a regular single-edge razor blade (Personna) after injection with 1mL of ice-cold 1 ⁇ HBSS (Gibco) using a 3-inch 18G needle (Air-Tite) and 5mL syringe (BD).
  • Dispersed tissues were aspirated into a 5mL syringe via a 3-inch 18G needle and transferred to a 15mL conical.5mL of ice-cold 1 ⁇ HBSS (Gibco) was added, and cells were mechanically dissociated by the oscillation of the volume through the syringe and needle for 20 cycles.
  • U1197.70243WO00 139/193 #13587456v2 suspension was centrifuged at 380 x g for 5 minutes at 4°C.1mL ice cold ACK Lysis Buffer (Gibco) was added to bone marrow cells, leukocytes, and splenocyte to resuspend post centrifugation for 5 minutes and subsequently diluted with 5mL ice-cold FACS Buffer (10% FBS in 1 ⁇ HBSS) then strained through a 40 ⁇ m cell strainer. Cells from each tissue were isolated by centrifugation at 380 x g for 5 minutes at 4°C.
  • leukocytes were repeatedly cycled, up to four additional times, through ACK lysis buffer (Gibco) and FACS buffer wash as previously described. Viability was manually determined by cell count using a standard trypan blue (Corning) exclusion method. [0470] Single-cell suspensions were prepared as described in the above sections. Samples were stained with viability dye (Violet, Invitrogen) in 1 ⁇ PBS pH 7.4 (Gibco) at RT protected from light for 15 minutes. Cells were resuspended and washed with FACS buffer (10% FBS, 1 ⁇ HBSS) and stored on ice until analysis.
  • viability dye Violet, Invitrogen
  • CCR2 RFP/WT /CX3CR1 GFP/WT bone marrow cells were derived into M-MDSCs ex vivo as described herein (CCR2 RFP+ /CX3CR1 GFP+ ).
  • Cells were washed with FACS buffer (10% FBS, 1 ⁇ HBSS) and resuspended in serum-free 1 ⁇ HBSS (Gibco). Viability and concentration were determined by trypan blue exclusion. Cells were adjusted to a concentration of ⁇ 1 ⁇ 10 6 /mL in 1.5mL microcentrifuge tubes (Fisher Scientific) and centrifuged at 500 x g for 5 minutes at 4°C.
  • Cells were resuspended in 400 ⁇ L of dissolved dendrimer solution at a concentration range from 1-100 ⁇ g/mL in 1 ⁇ HBSS or 1 ⁇ PBS at room temperature and incubated for 30 minutes protected from light. Samples were then washed in 1 ⁇ HBSS, stained for viability, and resuspended with FACS buffer in three technical repeats and analyzed via Spectral Flow Cytometry as described herein.
  • dendrimer stock solutions were prepared by fully solubilizing dendrimers in 1 ⁇ PBS pH 7.4 (Gibco), followed with filtration through a 13mm 0.22 ⁇ m PES syringe filter (Cytiva). Solutions were diluted at room temperature to concertation of 0.86 mg/mL in either competent or heat-inactivated (60°C for 30 minutes) sex pooled, complement preserved, C57BL/6 murine serum (Charles River) in 1.5mL microcentrifuge tubes.
  • Dendrimer-serum stock solutions were then incubated at 37°C for 30 minutes and brought to room temperature before co-incubation with cells at escalating doses (5-100 ⁇ /mL) in three biological repeats.
  • the dendrimer uptake as indicated by Mean Fluorescence Intensity (MFI) was subjected to flow cytometry analysis by gating out the CCR2 RFP+ /CX3CR1 GFP+ (M-MDSC) population.
  • Brain (tumors), spleens, and femurs were excised, and connective tissues were removed prior to transfer to 5 mL of 4 % w/v PFA at 4°C for 1 hour (brains and spleens) or 72 hours (femurs) at 2–8°C. Brains (tumors) and spleens were then transferred to a 30% w/v sucrose (Fisher Scientific) in water (Corning) solution for ⁇ 24 hours in 15mL conical tubes stored at 2–8°C.
  • Femurs were subsequently transferred to 5 mL of decalcification solution (20% EDTA, 10N NaOH, pH 7.4) for 4 days at 2-8°C and then a 30% w/v sucrose solution for 24 hours at 2-8°C. All tissues were embedded in optimal cutting temperature compound (Fisher Scientific) and cryo-sectioned at 10 ⁇ m or 30 ⁇ m thick sections at -25°C. Sections were prepared by addition to microscopy slides (Fisher Scientific), washed for 3 repetitive cycles with cold 1 ⁇ Dulbecco’s Phosphate Buffered Saline (DPBS) in a staining dish.
  • decalcification solution 20% EDTA, 10N NaOH, pH 7.4
  • DPBS Phosphate Buffered Saline
  • an anti-mouse CD31-Spark YG 570 labeled mAb (BioLegend) was added to hydrophobic pen (Vector Laboratories) encircled sections at a concentration of 10 ⁇ g/mL. Slides were then mounted with Vectashield anti-fade mounting medium with DAPI stain (Vector Laboratories) and coverslip. Slides were sealed with CoverGrip sealant and subsequently stored at 2- 8°C protected from light in the staining tray. Sections were analyzed at high magnification using an inverted Nikon A1R confocal microscope.
  • Widefield fluorescent images were generated using a Keyence BZ-X800 or Nikon Ti-E for fluorescence microscopy. Widefield fluorescence microscopy images were processed using Nikon Elements software v5.21 and confocal fluorescence microscopy using Fiji v2.9.0. [0473] Quantification of dendrimer concentration in plasma. Blood was collected from euthanized CCR2 RFP/WT /CX3CR1 GFP/WT mice bearing 3–4-week KR158 or GL261 gliomas at 24- or 72-hours post-dendrimer administration (tail vein) as previously described herein. Plasma samples were thawed from -80°C storage to room temperature and diluted 1:4 with 1 ⁇ PBS (Gibco).
  • Samples were then transferred to a 96-well clear bottom black plate (Thermo Scientific) and analyzed for absolute fluorescence intensity from Cy5 (635/675 (ex/em), integration: 400ms, read height: 3.0mm) using a Molecular Devices SpectraMax iD3 Multi-Mode Microplate Reader. Samples were plotted against a standard curve of Cy5 in murine serum and interpolated post-transgenic murine plasma background subtraction. The percentage of the injected dose was calculated by dilution factors ⁇ estimated dendrimer concentration ⁇ plasma volume (estimated to be 1.8 mL/mouse) / total injected dose. Samples were evaluated by 3 technical repeats.
  • Example 2 Dendrimer Targeting and Uptake Studies [0475] Introduction. The focus of nanoparticles in vivo trafficking has been mostly on their tissue- level biodistribution and clearance. Recent progress in the nanomedicine field suggests that the targeting of nanoparticles to immune cells can be used to modulate the immune response and enhance therapeutic delivery to the diseased tissue. In the presence of tumor lesions, monocytic-myeloid- derived suppressor cells (M-MDSCs) expand significantly in the bone marrow, egress into peripheral blood, and traffic solid tumor, where they help maintain an immuno-suppressive tumor microenvironment.
  • M-MDSCs monocytic-myeloid- derived suppressor cells
  • M-MDSCs in the tumor and lymphoid organs (bone marrow, spleen, peripheral blood, and tumor) can efficiently endocytose hydroxyl dendrimers.
  • the trafficking of M-MDSCs from the bone marrow to the tumor contributed to the deposition of hydroxyl dendrimers in the tumor.
  • M-MDSCs showed different capacities of endocytosing dendrimers of different functionalities in vivo.
  • M-MDSCs and microglia showed a high capacity of taking up hydroxyl dendrimers and these two cellular compartments accounted for more than half the amount of the hydroxyl dendrimer deposition in the tumor. This differential uptake and targeting was mediated by the unique serum proteins associated with each dendrimer surface functionality. [0477] M-MDSCs are recruited to the tumor from bone marrow. To determine how dendrimers interact with the M-MDSCs and other immune cells in vivo, the profiles of M-MDSCs and other infiltrative immune cells were first characterized in a GL261 mouse glioma model.
  • the GL261 glioma model was set up using CCR2 RFP/WT CX3CR1 GFP/WT transgenic mice, which allows the direct surveillance of the profile of infiltrative immune cells via the endogenously expressed red fluorescent protein (RFP) for chemokine receptor two (CCR2) and green fluorescent protein (GFP) for C-X3-C motif chemokine receptor 1 (CX3CR1) 18, 31 .
  • RFP red fluorescent protein
  • CCR2 chemokine receptor two
  • GFP green fluorescent protein
  • CX3CR1 CX3-C motif chemokine receptor 1
  • GPCRs G-Protein Coupled Receptors
  • U1197.70243WO00 142/193 #13587456v2 cells have been shown to suppress both CD4+ and CD8+ T cells in mouse glioma model 29 .
  • flow cytometry analysis of the M-MDSC population at the bone marrow was performed, which serves as the hematopoietic tissue of M-MDSCs; at the blood, which serves as the conduit to their destination; and at the spleen, which serves as a temporary reservoir.
  • M- MDSCs accounted for 11.4 ⁇ 0.4% and 3.8 ⁇ 0.1% of total cells in the bone marrow and spleen of GL261 tumor-bearing mice (FIGs.1A-1B).
  • M-MDSCs CCR2 and its cognate receptors mediated M-MDSCs egress from bone marrow into peripheral blood 29 , in which the M-MDSCs comprised 8.5 ⁇ 1.5% of the blood leukocytes (FIGs.1A-1B); M-MDSCs infiltrated the glioma through peripheral blood, ultimately comprising 22.1 ⁇ 1.0% of the stromal cells in the GL261 tumor (FIG.1C).
  • M- MDSCs were shown as the RFP and GFP co-localized cells, as indicated by the arrows in FIG.1D.
  • the CCR2 RFP/WT CX3CR1 GFP/WT transgenic mice also enabled profiling of other immune cell subsets in the glioma TME.
  • the CCR2-/CX3CR1+ subsets (16.7 ⁇ 3.0%, FIG.1C, abbreviated as microglia) were CD45 low /MHC+/F4/80+/CD11c-/CD11b medium , likely representing the CNS tissue-resident microglia;
  • the CCR2+/CX3CR1- subsets (4.4 ⁇ 2.0%, FIG.1C, abbreviated as CCR2+) were CD45+/MHCII+/F4/80-/CD11c-/CD11b low , likely representing other infiltrate myeloid cells originated outside of the CNS;
  • the CCR2-/CX3CR1- subsets (26.4 ⁇ 4.9%, FIG.1C, abbreviated as other cells) were a collection of tumor cells and other tumor stroma cells.
  • CXCR1 int tumor stromal cells
  • PAMAM OH dendrimer (Generation 6) was used as a model dendrimer to probe the dendrimer uptake capacity of different cell subsets within the glioma TME and the lymphoid organs such as bone marrow, spleen, and blood.
  • Mice with established GL261 glioma were injected systemically with OH dendrimers at 50mg/kg – a dose that has been well-tolerated in vivo 32 .
  • the OH dendrimer was fluorescently labeled with a minimal amount of Cy5 dye ( ⁇ 5% by wt%) 27 .
  • tumor M- MDSCs have a higher capacity for endocytosing OH dendrimers than other cell subsets within the GL261 tumor.
  • the composition of all dendrimer-positive cells was evaluated within the GL261 tumor by gating out the dendrimer-positive populations from the whole tumor stroma cells.
  • GL261 glioma model well-recapitulates the histology of glioma 30 , it is well- established that the GL261 glioma model, unlike human glioblastoma, is immunogenic 30, 33, 34 . Specifically, GL261 has high MHC I expression and a high tumor mutational load 33 and responds well to checkpoint inhibitors 34 . The dendrimer interactions with M-MDSC and other immune infiltrative cells were next characterized in an immunosuppressive KR158 model with lower populations of infiltrative M-MDSC 35, 36 .
  • CCR2-/CX3CR1- subsets which are a collection of tumor cells and other tumor stroma cells, showed a strong negative correlation with OH dendrimer deposition (R ⁇ 0.6).
  • R ⁇ 0.6 OH dendrimer deposition
  • the selective uptake of OH dendrimer by tumor-associated microglia/macrophages has been reported in previous studies 26, 27 .
  • the ontogeny difference between CNS-resident microglia and bone marrow-derived macrophages has led to their different functions in cancer and different responses to macrophage-targeting therapeutics 37, 38 , indicating the importance of analyzing the cell-
  • the immunogenic GL261 tumor has a ‘hotter’ tumor milieu with more infiltrative immune cells than the KR158 tumor. Since the infiltrative myeloid cells within the tumor can contribute to the tumor-accumulation of nanoparticles 41, 42 , it is possible that the higher OH dendrimer deposition in the GL261 tumor was associated with the higher amount of infiltrative immune cells in the GL261 tumor.
  • the KR158 tumor showed a mushroom-like crown on top of the brain, while the GL261 histology was more representative of the human glioma (FIG.7). The histological difference could also contribute to the differential dendrimer uptake.
  • the trafficking kinetics of M-MDSC contribute to the dendrimer accumulation in the tumor.
  • the production of M-MDSCs is accelerated in the bone marrow, from which these cells are directly recruited to the brain tumor through peripheral blood or indirectly from the spleen, which serves as the temporary reservoir of M-MDSCs 18, 29, 31 (FIG.3A).
  • the cellular uptake of OH dendrimers was quantified (MFI) by M-MDSCs located in bone marrow, spleen, peripheral blood, and tumor (GL261) at 24 hours after dendrimer injection (50mg/kg).
  • the blood M-MDSCs showed the highest dendrimer uptake (MFI 1371 ⁇ 494), likely because blood M-MDSCs can directly access the dendrimers in the circulation without the limitation of any tissue barriers.
  • M-MDSCs are constantly being recruited to the tumor in large amounts during tumor development, blood M-MDSCs may carry endocytosed dendrimer to the tumor while they infiltrate the tumor stroma.
  • the change of the dendrimer-positive M-MDSCs percentage was analyzed between the 24- and 72-hour window in two cohorts of mice.
  • EPR Enhanced Permeability and Retention
  • Circulating myeloid cells such as inflammation-associated monocytes and granulocytes, may be able to actively transport nanoparticles from the blood to the inflamed tissue when they infiltrate the inflamed tissue 12, 41, 44 .
  • This alternative mechanism was confirmed in the present disclosure in a mouse model of glioma, by showing that highly tumor-infiltrative M-MDSCs can contribute to the tumor accumulation of OH dendrimers. This phenomenon can be leveraged to design M-MDSC-targeting therapeutics for enhanced tumor delivery.
  • Dendrimer surface functionality affects their interactions with M-MDSC in vivo.
  • Dendrimer surface functionality can significantly affect their in vivo behaviors, such as absorption, distribution, metabolism, elimination (ADME), and toxicity 28, 45-47 .
  • NH2 dendrimers cannot efficiently access M-MDSCs but can be readily taken up by M- MDSCs. Nanoparticles need to efficiently cross the tissue barriers (e.g. the blood vessels and tissue extracellular matrix) before successfully ‘targeting’ the M-MDSCs located in the tissue stroma. How dendrimer surface functionality affects their abilities to ‘target’ M-MDSCs was determined by measuring the percentage of dendrimer-positive M-MDSCs in tissue (% dendrimer+ M-MDSCs).
  • the bone marrow and the tumor are the origin and the destination of M-MDSC recruitment, therefore were selected as the tissues of interest in this study.
  • the % dendrimer+ M-MDSCs was significantly lower for NH2 than other dendrimers in the bone marrow of both KR158 tumor-bearing mice and healthy control (FIG.4A).
  • NH2 dendrimers also targeted less M-MDSCs (12.3 ⁇ 5.3%) than SA (28 ⁇ 12.7%) and OH dendrimers (19.0 ⁇ 6.3%) (FIG.4B).
  • the lower cell- targeting of NH2 dendrimers was likely due to their lack of ability to cross tissue barriers 28 .
  • mice that received SA dendrimers the M-MDSCs compartment and the CX3CR1 int compartment (cells that potentially derived from M-MDSCs) accounted for a higher fraction (30.3%) of the dendrimer-positive cells when compared to mice that received OH dendrimers (20.5%) (FIG.4F).
  • the difference between SA and OH dendrimer uptake was further validated in the GL261 tumor model. Within the GL261 tumor stroma, M-MDSCs took up more dendrimers of both SA and OH than other cell subsets (FIG.4G).
  • the dendrimer uptake was the highest for NH2, followed by SA, and OH dendrimers (FIG.5C).
  • FIG.5C the dendrimer uptake was the highest for NH2, followed by SA, and OH dendrimers.
  • the uptake of NH2 dendrimers had a greater decrease after serum incubation than the OH and SA dendrimers (FIG.5F), indicating the serum protein had a greater influence in mediating the interaction of M-MDSCs with NH2 dendrimers.
  • Serum proteins can be classified into opsonin (enhance uptake) and dysopsonin (reduce uptake) 51 .
  • dendrimers were incubated with either competent ‘active’ serum or ‘heat-inactivated’ mouse serum under 60°C for 30min (FIG.5G), for NH2 dendrimers, heat-inactivation of serum proteins reduced NH2 dendrimer uptake in a dose-depended manner for up to 60% (FIG.5H), indicating serum proteins associated with NH2 dendrimers actively mediated their uptake by M-MDSCs.
  • NPs with ultra- small architectures such as dendrimers in the 1 – 20nm size range.
  • dendrimers Compared to large NPs, dendrimers have sizes similar to proteins and may interact with serum proteins in different stoichiometries and configurations 51 .
  • dendrimers carry payloads on their surfaces. Therefore, the properties of the surface payload will affect how dendrimers interact with M-MDSCs.
  • NH2 and SA dendrimers were used to represent dendrimers carrying drug molecules of acidic and basic properties and compared them with -OH dendrimers (control).
  • M-MDSCs suppress the anti-tumor immune response locally at the TME and globally at the lymphoid organs.
  • nanoparticles need to efficiently target these cells both locally and globally.
  • U1197.70243WO00 149/193 #13587456v2 enable direct surveillance of M-MDSCs, it was shown that that M-MDSCs can infiltrate glioma through peripheral blood in large amounts.
  • Systemically injected hydroxyl dendrimers efficiently target M-MDSCs located in bone marrow, peripheral blood, spleen, and tumor.
  • M- MDSCs and microglia showed high capacity for endocytosis of hydroxyl dendrimers, and these two cellular compartments accounted for more than half the amount of the hydroxyl dendrimer deposition in the tumor.
  • dendrimer showed greater tumor deposition and higher efficiency of M-MDSC-targeting than the KR158 glioma model.
  • the recruitment of M-MDSCs from bone marrow to tumor contributed to the tumor deposition of hydroxyl dendrimers.
  • the surface functionality of dendrimers affects their ability to target M-MDSCs in vivo. Although amine dendrimers had the highest capacity of being endocytosed by M-MDSCs, they could not access these cells as efficiently as hydroxyl or succinamic acid dendrimers, potentially due to the lack of ability to cross tissue barriers.
  • M-MDSCs took up succinamic acid dendrimers more efficiently than hydroxyl dendrimers.
  • serum proteins can affect how dendrimers interact with M-MDSCs.
  • the serum proteins associated with amine dendrimers significantly enhanced their uptake by M-MDSCs, while serum proteins associated with hydroxyl and succinamic acid dendrimers slightly reduced their uptake by M-MDSCs.
  • dendrimer-based drug conjugates carry drug payload on their surfaces, the payload molecular properties could affect the in vivo fate, such as cell- and tissue-targeting of the final dendrimer-drug conjugates.
  • Example 3 Materials and Methods for Dendrimer-Drug Conjugates for Delivery to M-MDSCs [0490] General Information.
  • the NMR spectra 1 H, 1 H-decoupled 13 C were recorded with TMS as the internal standard.
  • the coupling constants (J values) are given in hertz.
  • the LLL12 57 and bi- functional dendrimer 58 were synthesized from literature methods.
  • the G6-OH dendrimer was purchased from Dendritech Inc and used after evaporation of their stock solvents. [0491] General procedure for the synthesis of LLL12 derived N-substituted tert-butyl-5- oxopentanoate.
  • the obtained solution was evaporated to dryness under reduced pressure at rt, and the final product was dissolved in water and subjected further dialysis against water for 6 h by changing the water after every 2 h.
  • the obtained solution was lyophilized for 32 h to obtain dendrimer-drug conjugate 12 as orange solid (30 mg).
  • the resulting mixture was irradiated in microwave (Biotage microwave reactor) at 45 o C for 7 h. After completion (monitored by TLC), the reaction mixture was diluted with DMF (1 mL) and dialyzed against DMF for 8 h by changing the solvent after every 4 h. The collected solvent was evaporated to dryness and residue was dissolved in water, lyophilized to get the dendrimer-drug conjugate, which was further purified by column chromatography (Sephadex g-25) to get pure dendrimer-drug conjugate 17. [0508] Drug conjugated G6-OH dendrimer via carbamate linker (17).
  • Step-2 To a solution of LLL12-hydrazone 18 (45 mg, 0.1419 mmol, 1.0 eq) was added acetyl-azide NHS ester 19 (28 mg, 0.1419 mmol, 1.0 eq) and DIEA (74 uL, 55 mg, 0.426 mmol, 3.0 eq) at rt. The resulting reaction mixture was stirred until completion (5 h, monitored by TLC), after completion the reaction mixture was concentrated on high vacuum and purified by silica gel column chromatography (gradient elution of ethyl acetate and hexane, 30/70-50/50) to provide 20.
  • Example 4 Development of Dendrimer-Drug Conjugates for Delivery to M-MDSCs [0512]
  • a novel approach based on the dendrimer targeting and uptake studies disclosed herein was developed to systemically deliver LLL12 – a small molecule inhibitor of STAT3 – by formulating LLL12 into dendrimer-based LLL12 conjugates (D-LLL12). This approach is shown in FIG.14.
  • D-LLL12 was formulated with fast- (D-LLL12C, compound 17) and slow-release (D- LLL12S, compound 12) profiles by reacting LLL12-linkers with bi-functional dendrimers derived from hydroxyl surface dendrimers (G6 PAMAM-OH) over 5 steps.
  • the D-LLL12 conjugates were fully characterized by NMR and HPLC, and IC50 efficacy and toxicity studies were conducted in comparison with LLL12 (FIGs.15-16).
  • the highest absorption wavelength was identified to in order to determine the minimum detectable concentration of LLL12 by HPLC (FIGs.18A-18E).
  • THP-1 STAT3-Luc reporter cell line is cultured in RPMI I640; 10% FBS (Heat- inactivated), 100U/mL penicillin, 100 ⁇ g/mL streptomycin (1xP/S), 25mM HEPES along with 1.0mM Sodium pyruvate; Puromycin (1 ⁇ g/mL); 0.05mM 2-mercaptoethanol, puromycin 1 ⁇ g/mL in the incubator at 37 o C in 5% CO2.
  • FBS Heat- inactivated
  • penicillin 100 ⁇ g/mL
  • streptomycin (1xP/S streptomycin
  • 25mM HEPES 25mM HEPES along with 1.0mM Sodium pyruvate
  • Puromycin (1 ⁇ g/mL) 0.05mM 2-mercaptoethanol, puromycin 1 ⁇ g/mL in the incubator at 37 o C in 5% CO2.
  • a black clear bottom 96-well plate assay is used for IC50 using the suspension THP-1 STAT3-Luc cells (FIG.35).
  • Day 0 – Cells are seeded at 60,000 cells/well in 100 ⁇ l of media (RPMI 1640 - without phenol red, 25mM HEPES, 100U/mL penicillin, 100 ⁇ g/mL streptomycin (1xP/S). The plate is incubated for 4 hours.
  • the free drug LLL12 and its dendrimer drug conjugate formulations - D-LLL12C (17) (CS0177) and D-LLL12S (12) (CS0186) are added to the wells at concentrations ranging from 11.1 ⁇ g/mL ⁇ 3.7 ⁇ g/mL ⁇ 1.23 ⁇ g/mL ⁇ 0.4111 ⁇ g/mL ⁇ 0.137 ⁇ g/mL ⁇ 0.0456 ⁇ g/mL ensuring triplicates per concentration.
  • the plate is further incubated for 72 hours treatment period. Controls (set up in triplicates) – 1. Cells only (Background control); 2. Cells + 15ul (150 ⁇ g/mL) D-Luc (Endogenous STAT3 expression); 3.
  • the concentration of test compounds versus bioluminescence is plotted and the data is analyzed by GraphPad prism. [0519] Results.
  • the IC50 study demonstrates that free drug (LLL12) has highest potency with the lowest IC50 value of 0.305 ⁇ g/mL (FIG.36A).
  • the IC50 study was repeated in three independent experiments (FIG.36B). The average IC50 and the standard deviation for each formulation are shown in Table 2. Table 2. IC50 values for LLL12 and dendrimer-LLL12 conjugates.
  • THP-1 cells are cultured in RPMI I640; 10% FBS (Heat-inactivated), 100U/mL penicillin, 100 ⁇ g/mL streptomycin (1xP/S), 25mM HEPES; 0.05mM 2-mercaptoethanol in the incubator at 37 o C in 5% CO 2 .
  • FBS Heat-inactivated
  • 100U/mL penicillin 100 ⁇ g/mL streptomycin (1xP/S)
  • 25mM HEPES 25mM HEPES
  • 2-mercaptoethanol in the incubator at 37 o C in 5% CO 2 .
  • Experimental procedure A black clear bottom 96-well plate assay is used with the suspension THP-1 cells.
  • Day 0 – Cells are seeded at 60,000 cells/well in 100 ⁇ l of media (RPMI 1640 - without phenol red, 25mM HEPES, 100U/mL penicillin, 100 ⁇ g/mL streptomycin (1xP/S). The plate is incubated for 4 hours. The free drug LLL12 and its dendrimer drug conjugate formulations - 17 (CS0177) and 12 (CS0186) are added to the wells at concentrations ranging from 300 ⁇ g/mL ⁇ 0.0456 ⁇ g/mL ensuring triplicates per concentration. The plate is further incubated for 72 hours treatment period. Controls (set up in triplicates) – 1. No cells + CellTitre blue reagent (Background control); 2.
  • Untreated cells (Negative control); 3. Cells + 1% Triton X-100 (Positive control).
  • Day 4 – 1% Triton X-100 is added as a positive control to account for maximum cell death.
  • the plate is incubated for 3 hours at 37 o C in 5% CO2.
  • the fluorescence is recorded by the SpectraMax iD3 plate reader at 560/590nm.
  • the fluorescence values of the culture medium background are subtracted from all the experimental and control wells.
  • the concentration of test compounds versus % viability is plotted, and the data is analyzed by GraphPad prism.
  • Zhang F Xu Z, Jolly KJ. Myeloid cell-mediated drug delivery: From nanomedicine to cell therapy. Adv Drug Deliv Rev.2023;197:114827. Epub 20230415. doi: 10.1016/j.addr.2023.114827. PubMed PMID: 37068659.
  • Zhao Z Ukidve A, Kim J, Mitragotri S. Targeting Strategies for Tissue-Specific Drug Delivery. Cell.2020;181(1):151-67. doi: 10.1016/j.cell.2020.02.001. PubMed PMID: 32243788. [0533] 6.
  • Marvel D Gabrilovich DI. Myeloid-derived suppressor cells in the tumor microenvironment: expect the unexpected. J Clin Invest.2015;125(9):3356-64. Epub 20150713. doi: 10.1172/JCI80005. PubMed PMID: 26168215; PMCID: PMC4588239. [0536] 9. Gabrilovich DI. Myeloid-Derived Suppressor Cells. Cancer Immunology Research. 2017;5(1):3-8. doi: 10.1158/2326-6066.Cir-16-0297. PubMed PMID: WOS:000392240600001. [0537] 10.
  • Zhang F Mastorakos P, Mishra MK, Mangraviti A, Hwang L, Zhou J, Hanes J, Brem H, Olivi A, Tyler B, Kannan RM.
  • Wu LP Ficker M, Christensen JB, Simberg D, Trohopoulos PN, Moghimi SM. Dendrimer end-terminal motif-dependent evasion of human complement and complement activation through IgM hitchhiking. Nat Commun.2021;12(1):4858. Epub 20210811. doi: 10.1038/s41467-021- 24960-6. PubMed PMID: 34381048; PMCID: PMC8357934. [0585] 53. (a) Zhang, F.; Magruder, J. T.; Lin, Y.-A.; Crawford, T. C.; Grimm, J.
  • the invention includes embodiments in which more than one, or all of the group members are present in, employed in, or otherwise relevant to a given product or process.
  • the disclosure encompasses all variations, combinations, and permutations in which one or more limitations, elements, clauses, and descriptive terms from one or more of the listed claims is introduced into another claim.
  • any claim that is dependent on another claim can be modified to include one or more limitations found in any other claims that is dependent on the same base claim.
  • elements are presented as lists, e.g., in Markush group format, each subgroup of the elements is also disclosed, and any element(s) can be removed from the group.

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Abstract

The present disclosure provides compounds of the formulae herein (e.g., Formulae (I), (I'), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), (XI)), and pharmaceutically acceptable salts thereof, and compositions and kits comprising the compounds, or pharmaceutically acceptable salts thereof, which are useful for the delivery of agents to immune cells (e.g., myeloid-derived suppressor cells (MDSCs)). The present disclosure also provides methods of treating or preventing diseases or disorders (e.g., diseases or disorders associated with MDSCs) by administering the compounds, or pharmaceutically acceptable salts thereof, or compositions thereof, to a subject in need thereof, and methods of preparing the compounds, or pharmaceutically acceptable salts thereof.

Description

SELECTIVE DELIVERY OF AGENTS TO MYELOID-DERIVED SUPPRESSOR CELLS WITH A DENDRIMER-BASED FORMULATION CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This application claims the benefit of and priority under 35 U.S.C. § 119(e) to U.S. Provisional Application Number 63/627,659, filed January 31, 2024, titled SELECTIVE DELIVERY OF AGENTS TO MYELOID-DERIVED SUPPRESSOR CELLS WITH A DENDRIMER-BASED FORMULATION, the contents of which are incorporated herewith by reference in their entirety. BACKGROUND OF THE INVENTION [0002] Nanoparticles have been used to deliver therapeutic payload for disease treatment. While most in vivo biodistribution studies of nanoparticles have been focused on their tissue-level accumulation and clearance1, 2, recent progress in the nanomedicine field suggested that targeting nanoparticles to immune cells can be used to modulate the immune response and to enhance therapeutic delivery to the disease region3-5. [0003] For example, monocytic-myeloid-derived suppressor cells (M-MDSCs) are important cellular targets in cancer6. M-MDSCs are pathologically activated monocytes with potent immunosuppressive activities. Clinically, a high burden of M-MDSCs is associated with poor prognosis of many solid tumors7. In cancer, M-MDSCs help create and maintain an immunosuppressive tumor microenvironment (TME)8, 9. These cells can suppress anti-tumor T cells and promote regulatory T cells and anti-inflammatory myeloid cells8, 9. As such, there is an urgent need to develop delivery strategies to target M-MDSCs for cancer treatment. M-MDSCs are also associated with sepsis. [0004] In addition to their immunosuppressive features, M-MDSCs, as phagocytes, could also significantly affect the in vivo fate of nanoparticles. Historic studies have established the roles of myeloid cells in clearing nanoparticles10, 11, while emerging evidence has shown that myeloid cells in circulation can take up nanoparticles and actively transport them to the inflamed tissue4, 12, 13. M- MDSCs are significantly elevated in the peripheral blood of high-grade glioblastoma patients, accounting for as much as 10% of total cells in the peripheral blood and 30% of total peripheral blood mononuclear cells14, 15. In the presence of tumor lesions, the bone marrow accelerates monopoiesis and enhances the egress of M-MDSCs to the systemic circulation, leading to significant expansion of their population in the peripheral blood and in the spleen16. Tumor constantly recruits M-MDSCs in large amounts through CCR2-mediated chemotaxis to replenish the tumor-associated macrophages (TAMs)17, 18. Because of the abundance of M-MDSCs and their constant infiltration to the tumor sites, M-MDSCs have potential in mediating nanoparticle deposition at the tumors. [0005] Given the roles of M-MDSCs in establishing TME and mediating nanoparticle tumor- targeting, many efforts have tried to establish the correlation between nanoparticle physiochemical properties, such as size, surface charge, and targeting ligand to their targeting of M-MDSCs19-22.
U1197.70243WO00 1/193 #13587456v2 However, a significant challenge in studying the cell targeting behaviors of nanoparticles is that when nanoparticles are injected into the blood, multiple serum proteins such as immunoglobulins, fibrinogen, complement proteins, and apolipoproteins readily adsorb to the nanoparticle surface, forming a ‘protein corona’23, 24. The protein corona masks nanoparticle interactions with the cell surface and alters the nanoparticle’s cellular tropism23, 24. The nanoparticle-associated proteins dictate nanoparticle interactions with cells and more broadly their in vivo targeting behaviors, while the physical properties of nanoparticles may play a secondary role in this process. [0006] Dendrimers represent a class of ultra-small nanoparticles with sub-10nm size and carrying drug payload on their surfaces. About 26 dendrimer-based therapeutics with various types of payloads are currently under Phase I-III clinical trials25. Systemically administrated hydroxyl-terminated PAMAM dendrimer has been shown to selectively target TAMs in murine glioblastoma (glioma) models26, 27. However, given the heterogeneous nature of TAMs, it is unclear what subset(s) are being targeted and what mechanism mediates the selective cell-targeting. [0007] Signal transducer and activator of transcription 3 (STAT3) is a transcription factor highly activated in MDSCs that promotes angiogenesis, host immunosuppression, and tumor invasion. Clinical translation of STAT3 inhibitors has been hampered because of poor solubility and bioavailability, off-target effects, and limited efficacy in clinical trials. SUMMARY OF THE INVENTION [0008] The present disclosure provides an approach for the systemic delivery of agents, such as LLL12, a small molecule inhibitor of STAT3, by formulating the agents into dendrimer-based conjugates. [0009] In one aspect, the present disclosure provides compounds of Formula (I): and pharmaceutically acceptable salts thereof, wherein L, X, Y, Z, n, p, and q are as defined herein. [0010] In another aspect, the present disclosure provides a compound of Formula (I′): and pharmaceutically acceptable salts thereof, wherein L, X, Y, Z, n, p, and q are as defined herein.
U1197.70243WO00 2/193 #13587456v2 [0011] In another aspect, the present disclosure provides compounds of Formula (II): and pharmaceutically acceptable salts thereof, wherein L, Y, Zn, n, p, and q are as defined herein. [0012] In another aspect, the present disclosure provides compounds of Formula (III): and pharmaceutically acceptable salts thereof, wherein L, Y, Zn, n, p, and q are as defined herein. [0013] In another aspect, the present disclosure provides compounds of Formula (IV): and pharmaceutically acceptable salts thereof, wherein L, Y, Zn, n, p, and q are as defined herein. [0014] In another aspect, the present disclosure provides compounds of Formula (V): and pharmaceutically acceptable salts thereof, wherein L, Y, Zn, n, p, and q are as defined herein. [0015] In another aspect, the present disclosure provides compounds of Formula (VI): and pharmaceutically acceptable salts thereof, wherein L, Y, Zn, n, p, and q are as defined herein.
U1197.70243WO00 3/193 #13587456v2 [0016] In another aspect, the present disclosure provides compounds of Formula (VII): and pharmaceutically acceptable salts thereof, wherein L, Y, Zn, n, p, and q are as defined herein. [0017] In another aspect, the present disclosure provides compounds of Formula (VIII): and pharmaceutically acceptable salts thereof, wherein L, Y, Zn, n, p, and q are as defined herein. [0018] In another aspect, the present disclosure provides compounds of Formula (IX): and pharmaceutically acceptable salts thereof, wherein L, Y, Zn, n, p, and q are as defined herein.
U1197.70243WO00 4/193 #13587456v2 [0019] In another aspect, the present disclosure provides compounds of Formula (X): and pharmaceutically acceptable salts thereof, wherein L, Y, Zn, n, p, and q are as defined herein. [0020] In another aspect, the present disclosure provides compounds of Formula (XI): and pharmaceutically acceptable salts thereof, wherein L, Y, Zn, n, p, and q are as defined herein. [0021] In another aspect, the present disclosure provides a method of treating or preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formulae (I), (I′), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), or (XI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. [0022] In another aspect, the present disclosure provides a method of modulating a target protein, the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formulae (I), (I′), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), or (XI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. [0023] In another aspect, the present disclosure provides kits comprising a provided compound or provided composition and instructions for its use. [0024] It should be appreciated that the foregoing concepts, and the additional concepts discussed below, may be arranged in any suitable combination, as the present disclosure is not limited in this respect. Further, other advantages and novel features of the present disclosure will become apparent from the following detailed description of various non-limiting embodiments when considered in conjunction with the accompanying drawings.
U1197.70243WO00 5/193 #13587456v2 BRIEF DESCRIPTION OF THE DRAWINGS [0025] FIGs.1A-1D show that CCR2RFP/WTCX3CR1GFP/WT transgenic mice enable direct surveillance of M-MDSCs in mouse glioma model. FIG.1A shows that in mice with established GL261 tumors, the pie graph shows the flow cytometry measurement of the average percentage of M-MDSCs that reside in the reservoir tissues such as bone marrow, spleen, and peripheral blood leukocytes. At 3-4 weeks after tumor initiation, tissues from 6 tumor-bearing mice were analyzed. FIG.1B shows a gating strategy for the M-MDSCs cells in each tissue: the M-MDSCs are defined as the CCR2+/CX3CR1+ population. FIG.1C shows the gating strategy and the average percentage of major cell subsets in the GL261 tumor stroma. CCR2+/CX3CR1+ cells: M-MDSCs; CCR2- /CX3CR1+ cells: likely representing the CNS tissue-resident microglia; CCR2-/CX3CR1meidum cells: CX3CR1int, likely represent immune cells infiltrate brain tumor from external sources; CCR2+/CX3CR1- cells: CCR2+, likely represent other infiltrate myeloid cells originated outside of the CNS. CCR2-/CX3CR1- cells: other cells, a collection of tumor cells and other tumor stroma cells. At 3-4 weeks after tumor initiation, tissues from 6 tumor-bearing mice were analyzed. FIG.1D shows a confocal microscopy image of tumor (KR158). Arrow indicates the CCR2+/CX3CR1+ M-MDSCs.. [0026] FIGs.2A-2H show that tumor M-MDSCs efficiently endocytose dendrimers with high capacity. Cy5-labeled OH dendrimers (50mg/kg) were systemically injected into GL261 and KR158 tumor bearing transgenic mice (CCR2WT/RFPCX3CR1WT/GFP) at 3-4 weeks post-implantation; at 24 hours post injection, OH dendrimers uptake in myeloid subsets (indicated by Cy5 Median Fluorescence Intensity (MFI)) and the composition of dendrimer-positive cells (indicated by percentage of dendrimer-positive subset) within the tumor were analyzed through flow cytometry. FIG.2A shows the capacity of each cell subsets to uptake OH dendrimers within the GL261 gliomas at 24 hours post-injection. This is indicated by the MFI, which representatively measures the median number of dendrimers deposited per single cell. FIG.2B shows the statistical analysis of FIG.2A. FIG.2C shows a representative density plot of GL261 tumor stromal cells (left panel) and in the same tumor, the density plot of OH dendrimer-positive cells (right panel). M-MDSCs are shown in the box. FIG.2D shows the statistical analysis of the density plot of dendrimer-positive cells in FIG. 2C (right panel), which shows the composition of dendrimer-positive cells in each cellular compart within the GL261 tumor as a percent of a whole (n=6). FIG.2E shows the comparison of the mean, upper and lower quartiles of OH dendrimer deposition in tumor as measured by MFI between GL261 and KR158 gliomas. FIG.2F shows the comparison of OH dendrimer uptake capacity (MFI) in each cell subsets between GL261 and KR158 tumors. FIG.2G shows the composition of dendrimer- positive cells in each cellular compart within the KR158 tumor as a percent of a whole (n=6). FIG. 2H shows the correlation analysis between the abundance of cell subset (indicated by the percentage of cell subset within all tumor stroma cells) and the dendrimer deposition within the tumor (indicated by Cy5 MFI of all tumor cells) for both GL261 and KR158 tumors. The Pearson correlation coefficient (R) is based on 95% confidence interval. Weak correlation, R>0.2 or R<-0.2; strong
U1197.70243WO00 6/193 #13587456v2 correlation, R>0.6 or R<-0.6. For all in vivo experiment, data were generated based on 6 mice of both male and female sexes, *p<0.05, **p<0.01, ***p<0.001, ns=not statistically significant. [0027] FIGs.3A-3E show the trafficking kinetics of M-MDSC contributes to the dendrimer accumulation in the tumor. FIG.3A shows a graphical illustration shows the trafficking kinetics of M-MDSCs. M-MDSCs are recruited to the brain tumor from bone marrow (hematopoiesis organ) or spleen (temporary reservoir) through systemic circulation (blood). FIG.3B shows a heatmap that shows the OH dendrimer uptake capacity (indicated by Cy5 MFI) for M-MDSCs in bone marrow, spleen, blood, and tumor in GL261 glioma bearing mice (average MFI from 6 mice). FIG.3C shows a histogram that shows the overall OH dendrimer uptake capacity (indicated by Cy5 MFI) for white blood cells isolated from the peripheral blood of GL261 tumor-bearing mice. Control cells (no dendrimer injection). Cell subsets (lymphocytes, monocytes, granulocytes) were gated based on the scattered plot FSC vs. SSC. FIG.3D shows a comparison of the percentage of dendrimer-positive M- MDSCs between 24 hours vs.72 hours after dendrimer injection. Data was obtained from GL261 tumor-bearing mice. FIG.3E shows quantification of OH dendrimer concentrations in the plasma of GL261 tumor-bearing C57BL/6 mice at 24 hours (n=7) and 72 hours (n=12) post-dendrimer injection. For all other experiments in this figure, flow cytometry analyses were based on 6 GL261 tumor- bearing mice that received systemic injection of 50 mg/kg OH dendrimers. *p<0.05, **p<0.01. [0028] FIGs.4A-4J show that dendrimer surface chemistry affects their interactions with M-MDSC in vivo. To determine how dendrimer surface chemistry affect their interaction with M-MDSCs in vivo, dendrimers with different terminal groups (i.e. Succinamic acid: SA, hydroxyl: OH, and amine: NH2) were systemically injected into healthy (n=3) or tumor bearing mice (n=4) at tolerable doses (50mg/kg for SA and OH, 10mg/kg for NH2). At 24 hours after dendrimer injection, the bone marrow and tumor were isolated for flow cytometry analysis. FIG.4A shows a comparison of the percentage of dendrimer-positive M-MDSCs (% dendrimer+ M-MDSCs) in the bone marrow of healthy (left) and KR158 tumor-bearing mice (right) between SA, OH, and NH2 dendrimers. The % dendrimer+ M- MDSCs measure dendrimer’s ability to ‘target’ M-MDSCs in the tissue. FIG.4B shows comparison of the percentage of dendrimer-positive M-MDSCs in the KR158 tumor between SA, OH, and NH2 dendrimers. FIG.4C shows representative histograms comparing the Cy5 MFI of dendrimer-positive cells for OH, SA, and NH2 dendrimers in the bone marrow of KR158 tumor-bearing mice (top) and healthy mice (bottom) respectively. FIGs.4D-4E show a comparison of the M-MDSC’s capacity to endocytose SA, OH, and NH2 dendrimers (indicated by MFI). Plot shows M-MDSCs isolated from the bone marrow of healthy mice (left) and KR158 tumor bearing mice (right) (FIG.4D) and the KR158 tumors (FIG.4E). FIG.4F shows a comparison of SA and OH dendrimers for the composition of dendrimer-positive cells within the KR158 tumor. FIG.4G shows the heatmap shows the uptake capacity of SA and OH dendrimers (indicated by Cy5 MFI) by different cell subsets within the GL261 tumor. Data displayed is median. FIGs.4H-4I show statistical analysis comparing the SA and OH dendrimers in terms of their uptake capacity (indicated by Cy5 MFI of dendrimer-positive
U1197.70243WO00 7/193 #13587456v2 cells) by M-MDSCs within the tumor (FIG.4H) and bone marrow (FIG.4I) of the GL261 tumor bearing mice. FIG.4J shows a comparison of SA and OH dendrimers for the composition of dendrimer-positive cells within the GL261 tumor. For all statistical analyses, *p<0.05, **p<0.01, ****p<0.0001. [0029] FIGs.5A-5I show the serum proteins associated with dendrimers dictate the interaction between dendrimers and M-MDSCs. FIG.5A shows a schematic illustration of the generation of M- MDSC from the bone marrow of CCR2WT/RFPCX3CR1WT/GFP transgenic mice. To generate the M- MDSCs, bone marrow cells were isolated and were cultured in KR158 conditioned media for 5 days. FIG.5B shows flow cytometry analysis shows that exposure of bone marrow cells to KR158 conditioned media for 5 days enriched the CCR2+/CX3CR1+ cells (M-MDSCs) from less than 10% to approximately 59%. FIG.5C shows the histogram compares the uptake of OH, SA, and NH2 dendrimers (indicated by Cy5 MFI) by ex vivo generated M-MDSCs in the absence of mouse serum. To generate the plot, M-MDSCs were incubated with dendrimers at room temperature (RT) for 30 minutes. FIG.5D shows a comparison of ζ-potentials (zeta potentials) (mV) of serum proteins (triangle) and NH2, OH, and SA dendrimers before (circle) and after incubation with mouse serum (square). Dendrimers were incubated at 0.86 mg/mL in normal murine serum at 37°C for 30 minutes. ζ-potentials (mV) determined using DLS. The experiment was repeated for 3 times (n=3). FIG.5E shows a schematic illustration of the experiment flow that determines the influence of serum proteins on the dendrimer uptake by M-MDSCs. FIG.5F shows the plot based on the experiment flow in FIG.5E, which shows dose-dependent dendrimer uptake (indicated by Cy5 MFI) by M-MDSCs after co- incubation with either PBS (solid line) or mouse serum (dotted line) for NH2 (circle), OH (square), and SA (triangle) dendrimers. Each date point is an average of 3 independent experiment. FIG.5G shows a schematic illustration of the experiment flow that determines how heat-inactivation of serum affect the uptake of NH2, OH, and SA dendrimers by M-MDSCs. FIG.5H shows the plot based on the experiment flow in (FIG.5G), which shows the fold change of dendrimer uptake (x-axis) before and after heat-inactivation of mouse serum as a function of dendrimer dose (y-axis). NH2: circle, OH: square, SA: triangle. Negative fold change indicates heat inactivation of serum decreased dendrimer uptake. Positive fold change indicates heat inactivation of serum enhanced dendrimer uptake. Each data point is an average of 3 independent experiment. FIG.5I shows a representative scattered plot of heat inactivation of mouse serum enhanced the uptake of OH dendrimers by M-MDSCs. **p<0.01, ***p<0.005. [0030] FIG.6A shows the capacity of each cell subsets to uptake OH dendrimers within the KR158 gliomas at 24 hours post-injection. This is indicated by the MFI, which representatively measures the median number of dendrimers deposited per single cell. FIG.6B shows the statistical analysis of FIG.6A. FIG.6C shows confocal microscopy image of dendrimer (white) distribution within the tumor (KR158) at 24 hours postinjection. Arrow and star indicate the intracellular localization of OH dendrimer with CCR2+/CX3CR1+ M-MDSCs.
U1197.70243WO00 8/193 #13587456v2 [0031] FIG.7 shows representative fluorescence image of GL261 and KR158 tumor. Arrow indicates the tumor stroma. [0032] FIG.8A shows murine bone marrow sagittal cross section showing OH dendrimer (Cy5) signal 24 hours post 50mg/kg injection of (OH) dendrimer. Image generated by fluorescence microscopy 4X magnification panel and optimized for brightness and contrast using Fiji. Box highlighting image zoom panel. Below showing OH dendrimer in the monocyte rich red marrow of spongy bone (circle). FIG.8B shows murine spleen sagittal cross section showing dendrimer (Cy5) signal 24 hours post 50mg/kg injection of OH dendrimer. Image generated by fluorescence microscopy 10X magnification panel. Box highlighting image zoom panel. Below showing dendrimer deposition in monocyte rich red pulp and exclusion from lymphocyte white pulp zones. The Cy5 labelling of the G6 PAMAM dendrimer was carried out by following previously published work53. The amine surface G6 PAMAM dendrimer (50 mg) was dissolved in borate buffer (2 mL, pH 8.5) at room temperature. The reaction mixture was cooled to 0 ℃and Cy5 mono NHS ester (3.6 eq, 2.5 mg) in DMSO (1 mL) was added. The reaction mixture was allowed to stir overnight and lyophilized. The obtained crude product was dissolved in water and dialyzed (membrane MWCO = 12-14 kDa) against pure DI water for 24 h with successive change of water every 3 h. The collected water in dialysis bag was lyophilized to get the amine surface D-Cy5 (48 mg). The Cy5 labelling on hydroxyl surface dendrimer was conducted using bi-functional hydroxyl surface dendrimer by following the above procedure. The hydroxyl surface bi-functional dendrimer and acid surface bi-functional dendrimer were synthesized using literature method54. The acid surface bi-functional dendrimer (0.000590 mmol, 1.0 eq, 50 mg) was dissolved in DMSO (2 mL) at room temperature, to this solution was added Cy5 NHS ester (0.0022 mmol, 3.66 eq, 1.72 mg) and DIEA (0.0065 mmol, 11 eq, 1.5 uL). The resulting reaction mixture was allowed to stir overnight and dialyzed against DMSO for 8 h by changing the solvent at least two times, which was further dialyzed against DMF for 12 h by changing the solvent two times. The collected solvent was evaporated under high vacuum and compound was dissolved in water, dialyzed against pure DI water for 6 h by changing the solvent every 2 hours. The collected water was lyophilized for 24 h to get pure acid surface D-Cy5 (45 mg). [0033] FIGs.9A-9B show flow cytometry analyses of dendrimer uptake in blood leukocytes. FIG. 9A shows a comparison of OH dendrimer uptake (MFI) in leukocyte subpopulations of GL261 and KR158 tumor-bearing mice. n=6. FIG.9B shows a comparison of the percent of dendrimer positive leukocytes by subpopulations of GL261 and KR158 tumorbearing mice. n=6. *p<0.05, **p<0.001, ***p=0.0001, ****p<0.0001. [0034] FIGs 10A-10B show the number-average mean-based size (FIG.10A) and z-potential (mV) (FIG.10B) of hydroxyl (OH), succinamic acid (SA), and amine (NH2) dendrimers. The size and z- potential were measured by dynamic light scattering (DLS) and electrophoretic light scattering (ELS). FIG.10C shows a schematic illustration of the labeling of Cy5 to NH2 (top), OH (middle), and SA (bottom) dendrimers.
U1197.70243WO00 9/193 #13587456v2 [0035] FIG.11A shows in vitro evaluation of the dose-dependent toxicity of NH2, OH, and SA dendrimers on primary M-MDSCs by cell-titer blue assay. Positive control: cells treated with 1% Triton-X, Negative control: cell without treatment. FIG.11B shows that at 24 hours after systemic injection, NH2 dendrimers (10mg/kg) co-localized with the endothelial cell (indicated by arrow) in tumor stroma of a KR158 glioma established from CCR2WT/RFPCX3CR1WT/GFP transgenic mouse. [0036] FIG.12A shows NH2 dendrimer distribution within the KR158 tumor at 24 hours post- injection (10mg/kg), Image based on the widefield fluorescence microscopy (left panel) with zoom (right panel). FIG.12B shows OH dendrimer distribution within the KR158 tumor at 24 hours post- injection (50mg/kg), Image based on widefield fluorescence microscopy (left panel) with zoom (right panel). FIG.12C shows SA dendrimer distribution within the KR158 tumor at 24 hours post-injection (50mg/kg), Image based on widefield fluorescence microscopy (left panel) with zoom (right panel). FIG.12D shows 10X merge confocal of tumor from FIG.12A. FIG.12E shows 10X merge confocal of tumor from tumor shown in FIG.12B. FIG.12F shows 10X merge confocal of tumor from tumor shown in FIG.12C. FIG.12G shows 60X merge confocal for amine dendrimer tumor shown in FIG. 12D. FIG.12H shows 60X merge confocal for amine dendrimer tumor shown in FIG.12E. FIG.12I shows 60X merge confocal for amine dendrimer tumor shown in FIG.12F. [0037] FIGs.13A-13B show the comparison of overall tumor depositions of SA and OH dendrimers (indicated by Cy5 MFI) in KR158 tumor (FIG.13A) and GL261 tumor (FIG.13B). NS: no statistical significance. [0038] FIG.14 shows a schematic of delivery of LLL12 via a dendrimer drug conjugate. [0039] FIG.15 shows a schematic of characterization of the dendrimer drug conjugate by dynamic light scattering (DLS). [0040] FIG.16 shows a schematic of characterization of drug release quantification using high pressure liquid chromatography (HPLC). [0041] FIG.17 shows the zeta potential of PAMAM dendrimers (mV) using electrophoretic light scattering in 10 mM NaCl at 0.03 mg/mL. Positive (NH3) refers to a PAMAM dendrimer with positively charged amine surface groups. Neutral (OH) refers to a PAMAM dendrimer with neutral hydroxyl surface groups. Negative (SA) refers to a PAMAM dendrimer with negatively charged succinamic acid surface groups. [0042] FIGs.18A-18H show HPLC studies of G6 PAMAM OH dendrimer at 210 nm (FIG.18A), LLL12 at 250 nm (FIG.18B), LLL12-linker at 250 nm (FIG.18C), dendrimer-LLL12 conjugate 12 at 250 nm (FIG.18D), dendrimer-LLL12 conjugate 17 at 250 nm (FIG.18E), dendrimer-LLL12 conjugate 17 release profile at pH 4.5 at 250 nm (FIG.18F), dendrimer-LLL12 conjugate 17 release profile at 210 nm (FIG.18G), and dendrimer-LLL12 conjugate 17 release profile at pH 7.4 at 250 nm (FIG.18H). [0043] FIGs.19A-19G show IC50 efficacy and toxicity studies. FIG.19A shows a schematic for the IC50 studies. FIG.19B shows a THP-1STAT3-LucIL-6 response curve. FIG.19C shows a dose response
U1197.70243WO00 10/193 #13587456v2 curve in THP-1STAT3-Luc. FIGs.19D-19E show the LLL12 IC50 mass. FIG.19F shows the IC50 for dendrimer-LLL12 conjugate 17. FIG.19G shows a comparison of toxicity between LLL12 and dendrimer-LLL12 conjugate 17. [0044] FIGs.20A-20B show a UV-vis spectrum of LLL12 (FIG.20A) and a calibration curve (FIG. 20B). [0045] FIGs.21A-21B show 1H NMR (FIG.21A) and 13C NMR (FIG.21B) spectra of 1. [0046] FIGs.22A-22B show 1H NMR (FIG.22A) and 13C NMR (FIG.22B) spectra of 3. [0047] FIGs.23A-23B show 1H NMR (FIG.23A) and 13C NMR (FIG.23B) spectra of 4. [0048] FIG.24 shows 1H NMR spectrum of 6. [0049] FIG.25 shows 1H NMR spectrum of 9. [0050] FIG.26 shows 1H NMR spectrum of 10. [0051] FIG.27 shows 1H NMR spectrum of 12. [0052] FIGs.28A-28B show 1H NMR (FIG.28A) and 13C NMR (FIG.28B) spectra of 14. [0053] FIG.29 shows 1H NMR spectrum of 16. [0054] FIG.30 shows 1H NMR spectrum of 17. [0055] FIGs.31A-31B show 1H NMR (FIG.31A) and 13C NMR (FIG.31B) spectra of 18. [0056] FIGs.32A-32B show 1H NMR (FIG.32A) and 13C NMR (FIG.32B) spectra of 20. [0057] FIGs.33A-33B show 1H NMR spectra of 21 from 0 ppm to 14 ppm (FIG.33A) and 0 ppm to 9 ppm (FIG.32B). [0058] FIGs.34A-34B show (FIG.34A) and Zeta Potential (FIG.34B) of dendrimer conjugates of the present disclosure, along with unfunctionalized G6-OH. Size and Zeta Potential were calculated in 10mM NaCl. [0059] FIG.35 shows a schematic of efficacy (IC50) and toxicity analyses for dendrimer-drug conjugates. [0060] FIGs.36A-36B show representative IC50 curves for free drug (LLL12), dendrimer-LLL12 conjugates D-LLL12C (17), D-LLL12S (12), and D- LLL12H (21) (FIG.36A) and average IC50 values from 3 repeated experiments (FIG.36B). [0061] FIGs.37A-37C show toxicity (measured by cell viability) for 17 (FIG.37A), 12 (FIG.37B), and 21 (FIG.37C), with LLL12 as a control. [0062] FIGs.38A-38E show the therapeutic window. FIGs.38A-38D show the overlays of the IC50 curve (bioluminescence; curve with open circles) and the viability curve (curve with shaded triangles) for LLL12 (FIG.38A), G6-LLL12C (D-LLL12C, 17) (FIG.38B), G6-LLL12S (D-LLL12S, 12) (FIG.38C), and G6-LLL12H (D-LLL12H, 21) (FIG.38D). FIG.38E shows the calculated therapeutic windows.
U1197.70243WO00 11/193 #13587456v2 DEFINITIONS [0063] Definitions of specific functional groups and chemical terms are described in more detail below. The chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75th Ed., inside cover, and specific functional groups are generally defined as described therein. Additionally, general principles of organic chemistry, as well as specific functional moieties and reactivity, are described in Thomas Sorrell, Organic Chemistry, University Science Books, Sausalito, 1999;Michael B. Smith, March’s Advanced Organic Chemistry, 7th Edition, John Wiley & Sons, Inc., New York, 2013; Richard C. Larock, Comprehensive Organic Transformations, John Wiley & Sons, Inc., New York, 2018; and Carruthers, Some Modern Methods of Organic Synthesis, 3rd Edition, Cambridge University Press, Cambridge, 1987. [0064] Compounds described herein can comprise one or more asymmetric centers, and thus can exist in various stereoisomeric forms, e.g., enantiomers and/or diastereomers. For example, the compounds described herein can be in the form of an individual enantiomer, diastereomer or geometric isomer, or can be in the form of a mixture of stereoisomers, including racemic mixtures and mixtures enriched in one or more stereoisomer. Isomers can be isolated from mixtures by methods known to those skilled in the art, including chiral high pressure liquid chromatography (HPLC) and the formation and crystallization of chiral salts; or preferred isomers can be prepared by asymmetric syntheses. See, for example, Jacques et al., Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981); Wilen et al., Tetrahedron 33:2725 (1977); Eliel, E.L. Stereochemistry of Carbon Compounds (McGraw–Hill, NY, 1962); and Wilen, S.H., Tables of Resolving Agents and Optical Resolutions p.268 (E.L. Eliel, Ed., Univ. of Notre Dame Press, Notre Dame, IN 1972). The present disclosure additionally encompasses compounds as individual isomers substantially free of other isomers, and alternatively, as mixtures of various isomers. [0065] When a range of values (“range”) is listed, it encompasses each value and sub-range within the range. A range is inclusive of the values at the two ends of the range unless otherwise provided. For example “C1-6 alkyl” encompasses, C1, C2, C3, C4, C5, C6, C1–6, C1–5, C1–4, C1–3, C1–2, C2–6, C2–5, C2– 4, C2–3, C3–6, C3–5, C3–4, C4–6, C4–5, and C5–6 alkyl. [0066] The term “alkyl” refers to a radical of a straight-chain or branched saturated hydrocarbon group having from 1 to 20 carbon atoms (“C1–20 alkyl”). In some embodiments, an alkyl group has 1 to 12 carbon atoms (“C1–12 alkyl”). In some embodiments, an alkyl group has 1 to 10 carbon atoms (“C1–10 alkyl”). In some embodiments, an alkyl group has 1 to 9 carbon atoms (“C1–9 alkyl”). In some embodiments, an alkyl group has 1 to 8 carbon atoms (“C1–8 alkyl”). In some embodiments, an alkyl group has 1 to 7 carbon atoms (“C1–7 alkyl”). In some embodiments, an alkyl group has 1 to 6 carbon atoms (“C1–6 alkyl”). In some embodiments, an alkyl group has 1 to 5 carbon atoms (“C1–5 alkyl”). In some embodiments, an alkyl group has 1 to 4 carbon atoms (“C1–4 alkyl”). In some embodiments, an alkyl group has 1 to 3 carbon atoms (“C1–3 alkyl”). In some embodiments, an alkyl group has 1 to 2
U1197.70243WO00 12/193 #13587456v2 carbon atoms (“C1–2 alkyl”). In some embodiments, an alkyl group has 1 carbon atom (“C1 alkyl”). In some embodiments, an alkyl group has 2 to 6 carbon atoms (“C2-6 alkyl”). Examples of C1–6 alkyl groups include methyl (C1), ethyl (C2), propyl (C3) (e.g., n-propyl, isopropyl), butyl (C4) (e.g., n-butyl, tert-butyl, sec-butyl, isobutyl), pentyl (C5) (e.g., n-pentyl, 3-pentanyl, amyl, neopentyl, 3-methyl-2- butanyl, tert-amyl), and hexyl (C6) (e.g., n-hexyl). Additional examples of alkyl groups include n- heptyl (C7), n-octyl (C8), n-dodecyl (C12), and the like. Unless otherwise specified, each instance of an alkyl group is independently unsubstituted (an “unsubstituted alkyl”) or substituted (a “substituted alkyl”) with one or more substituents (e.g., halogen, such as F). In certain embodiments, the alkyl group is an unsubstituted C1–12 alkyl (such as unsubstituted C1–6 alkyl, e.g., −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 or s-Bu), unsubstituted isobutyl (i-Bu)). In certain embodiments, the alkyl group is a substituted C1–12 alkyl (such as substituted C1–6 alkyl, e.g., –CH2F, –CHF2, –CF3, – CH2CH2F, –CH2CHF2, –CH2CF3, or benzyl (Bn)). [0067] The term “haloalkyl” is a substituted alkyl group, wherein one or more of the hydrogen atoms are independently replaced by a halogen, e.g., fluoro, bromo, chloro, or iodo. “Perhaloalkyl” is a subset of haloalkyl, and refers to an alkyl group wherein all of the hydrogen atoms are independently replaced by a halogen, e.g., fluoro, bromo, chloro, or iodo. [0068] The term “heteroalkyl” refers to an alkyl group, which further includes at least one heteroatom (e.g., 1, 2, 3, or 4 heteroatoms) selected from oxygen, nitrogen, or sulfur within (e.g., inserted between adjacent carbon atoms of) and/or placed at one or more terminal position(s) of the parent chain. In certain embodiments, a heteroalkyl group refers to a saturated group having from 1 to 20 carbon atoms and 1 or more heteroatoms within the parent chain (“C1–20 heteroalkyl”). In certain embodiments, a heteroalkyl group refers to a saturated group having from 1 to 12 carbon atoms and 1 or more heteroatoms within the parent chain (“C1–12 heteroalkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 11 carbon atoms and 1 or more heteroatoms within the parent chain (“C1–11 heteroalkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 10 carbon atoms and 1 or more heteroatoms within the parent chain (“C1–10 heteroalkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 9 carbon atoms and 1 or more heteroatoms within the parent chain (“C1–9 heteroalkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 8 carbon atoms and 1 or more heteroatoms within the parent chain (“C1–8 heteroalkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 7 carbon atoms and 1 or more heteroatoms within the parent chain (“C1–7 heteroalkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 6 carbon atoms and 1 or more heteroatoms within the parent chain (“C1–6 heteroalkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 5 carbon atoms and 1 or 2 heteroatoms within the parent chain (“C1–5 heteroalkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 4 carbon
U1197.70243WO00 13/193 #13587456v2 atoms and 1or 2 heteroatoms within the parent chain (“C1–4 heteroalkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 3 carbon atoms and 1 heteroatom within the parent chain (“C1–3 heteroalkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 2 carbon atoms and 1 heteroatom within the parent chain (“C1–2 heteroalkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 carbon atom and 1 heteroatom (“C1 heteroalkyl”). In some embodiments, a heteroalkyl group is a saturated group having 2 to 6 carbon atoms and 1 or 2 heteroatoms within the parent chain (“C2-6 heteroalkyl”). Unless otherwise specified, each instance of a heteroalkyl group is independently unsubstituted (an “unsubstituted heteroalkyl”) or substituted (a “substituted heteroalkyl”) with one or more substituents. In certain embodiments, the heteroalkyl group is an unsubstituted C1–12 heteroalkyl. In certain embodiments, the heteroalkyl group is a substituted C1–12 heteroalkyl. [0069] The term “alkenyl” refers to a radical of a straight-chain or branched hydrocarbon group having from 1 to 20 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 1 to 20 carbon atoms (“C1-20 alkenyl”). In some embodiments, an alkenyl group has 1 to 12 carbon atoms (“C1–12 alkenyl”). In some embodiments, an alkenyl group has 1 to 11 carbon atoms (“C1–11 alkenyl”). In some embodiments, an alkenyl group has 1 to 10 carbon atoms (“C1–10 alkenyl”). In some embodiments, an alkenyl group has 1 to 9 carbon atoms (“C1–9 alkenyl”). In some embodiments, an alkenyl group has 1 to 8 carbon atoms (“C1–8 alkenyl”). In some embodiments, an alkenyl group has 1 to 7 carbon atoms (“C1–7 alkenyl”). In some embodiments, an alkenyl group has 1 to 6 carbon atoms (“C1–6 alkenyl”). In some embodiments, an alkenyl group has 1 to 5 carbon atoms (“C1–5 alkenyl”). In some embodiments, an alkenyl group has 1 to 4 carbon atoms (“C1–4 alkenyl”). In some embodiments, an alkenyl group has 1 to 3 carbon atoms (“C1–3 alkenyl”). In some embodiments, an alkenyl group has 1 to 2 carbon atoms (“C1–2 alkenyl”). In some embodiments, an alkenyl group has 1 carbon atom (“C1 alkenyl”). The one or more carbon-carbon double bonds can be internal (such as in 2-butenyl) or terminal (such as in 1-butenyl). Examples of C1–4 alkenyl groups include methylidenyl (C1), ethenyl (C2), 1-propenyl (C3), 2-propenyl (C3), 1-butenyl (C4), 2-butenyl (C4), butadienyl (C4), and the like. Examples of C1–6 alkenyl groups include the aforementioned C2-4 alkenyl groups as well as 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 (an “unsubstituted alkenyl”) or substituted (a “substituted alkenyl”) with one or more substituents. In certain embodiments, the alkenyl group is an unsubstituted C1-20 alkenyl. In certain embodiments, the alkenyl group is a substituted C1-20 alkenyl. In an alkenyl group, a C=C double bond for which the stereochemistry is not specified (e.g., −CH=CHCH3 or ) may be in the (E)- or (Z)-configuration.
U1197.70243WO00 14/193 #13587456v2 [0070] The term “heteroalkenyl” refers to an alkenyl group, which further includes at least one heteroatom (e.g., 1, 2, 3, or 4 heteroatoms) selected from oxygen, nitrogen, or sulfur within (e.g., inserted between adjacent carbon atoms of) and/or placed at one or more terminal position(s) of the parent chain. [0071] The term “alkynyl” refers to a radical of a straight-chain or branched hydrocarbon group having from 1 to 20 carbon atoms and one or more carbon-carbon triple bonds (e.g., 1, 2, 3, or 4 triple bonds) (“C1-20 alkynyl”). [0072] The term “heteroalkynyl” refers to an alkynyl group, which further includes at least one heteroatom (e.g., 1, 2, 3, or 4 heteroatoms) selected from oxygen, nitrogen, or sulfur within (e.g., inserted between adjacent carbon atoms of) and/or placed at one or more terminal position(s) of the parent chain. [0073] The term “carbocyclyl” or “carbocyclic” refers to a radical of a non-aromatic cyclic hydrocarbon group having from 3 to 14 ring carbon atoms (“C3-14 carbocyclyl”) and zero heteroatoms in the non-aromatic ring system. [0074] The term “heterocyclyl” or “heterocyclic” refers to a radical of a 3- to 14-membered non- aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“3–14 membered heterocyclyl”). [0075] 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 ^ electrons shared in a cyclic array) having 6–14 ring carbon atoms and zero heteroatoms provided in the aromatic ring system (“C6-14 aryl”). [0076] The term “heteroaryl” refers to a radical of a 5-14 membered monocyclic or polycyclic (e.g., bicyclic, tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10, or 14 ^ electrons shared in a cyclic array) having ring carbon atoms and 1–4 ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-14 membered heteroaryl”). [0077] The term “unsaturated bond” refers to a double or triple bond. [0078] The term “unsaturated” or “partially unsaturated” refers to a moiety that includes at least one double or triple bond. [0079] The term “saturated” or “fully saturated” refers to a moiety that does not contain a double or triple bond, e.g., the moiety only contains single bonds. [0080] Affixing the suffix “-ene” to a group indicates the group is a divalent moiety, e.g., alkylene is the divalent moiety of alkyl, alkenylene is the divalent moiety of alkenyl, heteroalkylene is the divalent moiety of heteroalkyl, and heteroalkenylene is the divalent moiety of heteroalkenyl. [0081] A group is optionally substituted unless expressly provided otherwise. The term “optionally substituted” refers to being substituted or unsubstituted. In certain embodiments, alkyl, alkenyl, heteroalkyl, and heteroalkenyl groups are optionally substituted. “Optionally substituted” refers to a group which is substituted or unsubstituted (e.g., “substituted” or “unsubstituted” alkyl, “substituted”
U1197.70243WO00 15/193 #13587456v2 or “unsubstituted” alkenyl, “substituted” or “unsubstituted” heteroalkyl, “substituted” or “unsubstituted” heteroalkenyl). In general, the term “substituted” means that at least one hydrogen present on a group is replaced with a permissible substituent, e.g., a substituent which upon substitution results in a stable compound, e.g., a compound which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, or other reaction. Unless otherwise indicated, a “substituted” group has a substituent at one or more substitutable positions of the group, and when more than one position in any given structure is substituted, the substituent is either the same or different at each position. The term “substituted” is contemplated to include substitution with all permissible substituents of organic compounds, and includes any of the substituents described herein that results in the formation of a stable compound. The present disclosure contemplates any and all such combinations in order to arrive at a stable compound. For purposes of this disclosure, heteroatoms such as nitrogen may have hydrogen substituents and/or any suitable substituent as described herein which satisfy the valencies of the heteroatoms and results in the formation of a stable moiety. The disclosure is not limited in any manner by the exemplary substituents described herein. [0082] Exemplary carbon atom substituents include halogen, −CN, −NO2, −N3, −SO2H, −SO3H, −OH, −ORaa, −ON(Rbb)2, −N(Rbb)2, −N(Rbb)3+X−, −N(ORcc)Rbb, −SH, −SRaa, −SSRcc, −C(=O)Raa, −CO2H, −CHO, −C(ORcc)2, −CO2Raa, −OC(=O)Raa, −OCO2Raa, −C(=O)N(Rbb)2, −OC(=O)N(Rbb)2, −NRbbC(=O)Raa, −NRbbCO2Raa, −NRbbC(=O)N(Rbb)2, −C(=NRbb)Raa, −C(=NRbb)ORaa, −OC(=NRbb)Raa, −OC(=NRbb)ORaa, −C(=NRbb)N(Rbb)2, −OC(=NRbb)N(Rbb)2, −NRbbC(=NRbb)N(Rbb)2, −C(=O)NRbbSO2Raa, −NRbbSO2Raa, −SO2N(Rbb)2, −SO2Raa, −SO2ORaa, −OSO2Raa, −S(=O)Raa, −OS(=O)Raa, −Si(Raa)3, −OSi(Raa)3 −C(=S)N(Rbb)2, −C(=O)SRaa, −C(=S)SRaa, −NRbbP(=O)(Raa)2, −NRbbP(=O)(ORcc)2, −NRbbP(=O)(N(Rbb)2)2, −P(Rcc)2, −P(ORcc)2, −P(Rcc)3+X−, −P(ORcc)3+X−, −P(Rcc)4, −P(ORcc)4, −OP(Rcc)2, −OP(Rcc)3+X−, −OP(ORcc)2, −OP(ORcc)3+X−, −OP(Rcc)4, −OP(ORcc)4, −B(Raa)2, −B(ORcc)2, −BRaa(ORcc), C1–20 alkyl, C1–20 perhaloalkyl, C1–20 alkenyl, C1–20 alkynyl, heteroC1–20 alkyl, heteroC1–20 alkenyl, heteroC1–20 alkynyl, C3-10 carbocyclyl, 3- 14 membered heterocyclyl, C6-14 aryl, and 5-14 membered heteroaryl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rdd groups; wherein X− is a counterion; or two geminal hydrogens on a carbon atom are replaced with the group =O, =S, =NN(Rbb)2, =NNRbbC(=O)Raa, =NNRbbC(=O)ORaa, =NNRbbS(=O)2Raa, =NRbb, or =NORcc; wherein: each instance of Raa is, independently, selected from C1–20 alkyl, C1–20 perhaloalkyl, C1–20 alkenyl, C1–20 alkynyl, heteroC1–20 alkyl, heteroC1–20alkenyl, heteroC1–20alkynyl, C3-10 carbocyclyl, 3-14 membered heterocyclyl, C6-14 aryl, and 5-14 membered heteroaryl, or two Raa groups are joined to form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl
U1197.70243WO00 16/193 #13587456v2 ring, wherein each of the alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rdd groups; each instance of Rbb is, independently, selected from hydrogen, −OH, −ORaa, −N(Rcc)2, −CN, −C(=O)Raa, −C(=O)N(Rcc)2, −CO2Raa, −SO2Raa, −C(=NRcc)ORaa, −C(=NRcc)N(Rcc)2, −SO2N(Rcc)2, −SO2Rcc, −SO2ORcc, −SORaa, −C(=S)N(Rcc)2, −C(=O)SRcc, −C(=S)SRcc, −P(=O)(Raa)2, −P(=O)(ORcc)2, −P(=O)(N(Rcc)2)2, C1–20 alkyl, C1–20 perhaloalkyl, C1–20 alkenyl, C1–20 alkynyl, heteroC1–20alkyl, heteroC1–20alkenyl, heteroC1–20alkynyl, C3-10 carbocyclyl, 3-14 membered heterocyclyl, C6-14 aryl, and 5-14 membered heteroaryl, or two Rbb groups are joined to form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rdd groups; each instance of Rcc is, independently, selected from hydrogen, C1–20 alkyl, C1–20 perhaloalkyl, C1–20 alkenyl, C1–20 alkynyl, heteroC1–20 alkyl, heteroC1–20 alkenyl, heteroC1–20 alkynyl, C3-10 carbocyclyl, 3-14 membered heterocyclyl, C6-14 aryl, and 5-14 membered heteroaryl, or two Rcc groups are joined to form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rdd groups; each instance of Rdd is, independently, selected from halogen, −CN, −NO2, −N3, −SO2H, −SO3H, −OH, −ORee, −ON(Rff)2, −N(Rff)2, −N(Rff)3+X−, −N(ORee)Rff, −SH, −SRee, −SSRee, −C(=O)Ree, −CO2H, −CO2Ree, −OC(=O)Ree, −OCO2Ree, −C(=O)N(Rff)2, −OC(=O)N(Rff)2, −NRffC(=O)Ree, −NRffCO2Ree, −NRffC(=O)N(Rff)2, −C(=NRff)ORee, −OC(=NRff)Ree, −OC(=NRff)ORee, −C(=NRff)N(Rff)2, −OC(=NRff)N(Rff)2, −NRffC(=NRff)N(Rff)2, −NRffSO2Ree, −SO2N(Rff)2, −SO2Ree, −SO2ORee, −OSO2Ree, −S(=O)Ree, −Si(Ree)3, −OSi(Ree)3, −C(=S)N(Rff)2, −C(=O)SRee, −C(=S)SRee, −SC(=S)SRee, −P(=O)(ORee)2, −P(=O)(Ree)2, −OP(=O)(Ree)2, −OP(=O)(ORee)2, C1–10 alkyl, C1–10 perhaloalkyl, C1–10 alkenyl, C1–10 alkynyl, heteroC1–10alkyl, heteroC1–10alkenyl, heteroC1– 10alkynyl, C3-10 carbocyclyl, 3-10 membered heterocyclyl, C6-10 aryl, and 5-10 membered heteroaryl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rgg groups, or two geminal Rdd substituents are joined to form =O or =S; wherein X− is a counterion; each instance of Ree is, independently, selected from C1–10 alkyl, C1–10 perhaloalkyl, C1–10 alkenyl, C1–10 alkynyl, heteroC1–10 alkyl, heteroC1–10 alkenyl, heteroC1–10 alkynyl, C3-10 carbocyclyl, C6-10 aryl, 3-10 membered heterocyclyl, and 3-10 membered heteroaryl, wherein
U1197.70243WO00 17/193 #13587456v2 each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rgg groups; each instance of Rff is, independently, selected from hydrogen, C1–10 alkyl, C1–10 perhaloalkyl, C1–10 alkenyl, C1–10 alkynyl, heteroC1–10 alkyl, heteroC1–10 alkenyl, heteroC1–10 alkynyl, C3-10 carbocyclyl, 3-10 membered heterocyclyl, C6-10 aryl, and 5-10 membered heteroaryl, or two Rff groups are joined to form a 3-10 membered heterocyclyl or 5-10 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rgg groups; each instance of Rgg is, independently, halogen, −CN, −NO2, −N3, −SO2H, −SO3H, −OH, −OC1–6 alkyl, −ON(C1–6 alkyl)2, −N(C1–6 alkyl)2, −N(C1–6 alkyl)3+X−, −NH(C1–6 alkyl)2+X−, −NH2(C1–6 alkyl) +X−, −NH3+X−, −N(OC1–6 alkyl)(C1–6 alkyl), −N(OH)(C1–6 alkyl), −NH(OH), −SH, −SC1–6 alkyl, −SS(C1–6 alkyl), −C(=O)(C1–6 alkyl), −CO2H, −CO2(C1–6 alkyl), −OC(=O)(C1–6 alkyl), −OCO2(C1–6 alkyl), −C(=O)NH2, −C(=O)N(C1–6 alkyl)2, −OC(=O)NH(C1–6 alkyl), −NHC(=O)( C1–6 alkyl), −N(C1–6 alkyl)C(=O)( C1–6 alkyl), −NHCO2(C1–6 alkyl), −NHC(=O)N(C1–6 alkyl)2, −NHC(=O)NH(C1–6 alkyl), −NHC(=O)NH2, −C(=NH)O(C1–6 alkyl), −OC(=NH)(C1–6 alkyl), −OC(=NH)OC1–6 alkyl, −C(=NH)N(C1–6 alkyl)2, −C(=NH)NH(C1–6 alkyl), −C(=NH)NH2, −OC(=NH)N(C1–6 alkyl)2, −OC(NH)NH(C1– 6 alkyl), −OC(NH)NH2, −NHC(NH)N(C1–6 alkyl)2, −NHC(=NH)NH2, −NHSO2(C1–6 alkyl), −SO2N(C1–6 alkyl)2, −SO2NH(C1–6 alkyl), −SO2NH2, −SO2C1–6 alkyl, −SO2OC1–6 alkyl, −OSO2C1–6 alkyl, −SOC1–6 alkyl, −Si(C1–6 alkyl)3, −OSi(C1–6 alkyl)3 −C(=S)N(C1–6 alkyl)2, C(=S)NH(C1–6 alkyl), C(=S)NH2, −C(=O)S(C1–6 alkyl), −C(=S)SC1–6 alkyl, −SC(=S)SC1–6 alkyl, −P(=O)(OC1–6 alkyl)2, −P(=O)(C1–6 alkyl)2, −OP(=O)(C1–6 alkyl)2, −OP(=O)(OC1–6 alkyl)2, C1–10 alkyl, C1–10 perhaloalkyl, C1–10 alkenyl, C1–10 alkynyl, heteroC1–10 alkyl, heteroC1–10 alkenyl, heteroC1–10 alkynyl, C3-10 carbocyclyl, C6-10 aryl, 3-10 membered heterocyclyl, or 5-10 membered heteroaryl; or two geminal Rgg substituents can be joined to form =O or =S; and each X− is a counterion. [0083] The term “halo” or “halogen” refers to fluorine (fluoro, −F), chlorine (chloro, −Cl), bromine (bromo, −Br), or iodine (iodo, −I). [0084] The term “hydroxyl” or “hydroxy” refers to the group −OH. The term “substituted hydroxyl” or “substituted hydroxy,” by extension, refers to a hydroxyl group wherein the oxygen atom directly attached to the parent molecule is substituted with a group other than hydrogen, and includes groups selected from −ORaa, −ON(Rbb)2, −OC(=O)SRaa, −OC(=O)Raa, −OCO2Raa, −OC(=O)N(Rbb)2, −OC(=NRbb)Raa, −OC(=NRbb)ORaa, −OC(=NRbb)N(Rbb)2, −OS(=O)Raa, −OSO2Raa, −OSi(Raa)3,
U1197.70243WO00 18/193 #13587456v2 −OP(Rcc)2, −OP(Rcc)3 +X−, −OP(ORcc)2, −OP(ORcc)3 +X−, −OP(=O)(Raa)2, −OP(=O)(ORcc)2, and −OP(=O)(N(Rbb))2, wherein X−, Raa, Rbb, and Rcc are as defined herein. [0085] Nitrogen atoms can be substituted or unsubstituted as valency permits, and include primary, secondary, tertiary, and quaternary nitrogen atoms. Exemplary nitrogen atom substituents include hydrogen, −OH, −ORaa, −N(Rcc)2, −CN, −C(=O)Raa, −C(=O)N(Rcc)2, −CO2Raa, −SO2Raa, −C(=NRbb)Raa, −C(=NRcc)ORaa, −C(=NRcc)N(Rcc)2, −SO2N(Rcc)2, −SO2Rcc, −SO2ORcc, −SORaa, −C(=S)N(Rcc)2, −C(=O)SRcc, −C(=S)SRcc, −P(=O)(ORcc)2, −P(=O)(Raa)2, −P(=O)(N(Rcc)2)2, C1–20 alkyl, C1–20 perhaloalkyl, C1–20 alkenyl, C1–20 alkynyl, hetero C1–20 alkyl, hetero C1–20 alkenyl, hetero C1– 20 alkynyl, C3-10 carbocyclyl, 3-14 membered heterocyclyl, C6-14 aryl, and 5-14 membered heteroaryl, or two Rcc groups attached to an N atom are joined to form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rdd groups, and wherein Raa, Rbb, Rcc and Rdd are as defined above. [0086] In certain embodiments, the substituent present on the nitrogen atom is a nitrogen protecting group (also referred to herein as an “amino protecting group”). Nitrogen protecting groups include −OH, −ORaa, −N(Rcc)2, −C(=O)Raa, −C(=O)N(Rcc)2, −CO2Raa, −SO2Raa, −C(=NRcc)Raa, −C(=NRcc)ORaa, −C(=NRcc)N(Rcc)2, −SO2N(Rcc)2, −SO2Rcc, −SO2ORcc, −SORaa, −C(=S)N(Rcc)2, −C(=O)SRcc, −C(=S)SRcc, C1–10 alkyl, C1–20 alkenyl, C1–20 alkynyl, hetero C1–20 alkyl, hetero C1–20 alkenyl, hetero C1–20 alkynyl, C3-10 carbocyclyl, 3-14 membered heterocyclyl, C6-14 aryl, and 5-14 membered heteroaryl groups, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rdd groups, and wherein Raa, Rbb, Rcc and Rdd are as defined 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. Wuts, 3rd edition, John Wiley & Sons, 1999, incorporated herein by reference. [0087] In certain embodiments, each oxygen atom substituent is independently substituted (e.g., substituted with one or more halogen) or unsubstituted C1-10 alkyl, −C(=O)Raa, −CO2Raa, −C(=O)N(Rbb)2, or an oxygen protecting group. In certain embodiments, each oxygen atom substituents is independently substituted (e.g., substituted with one or more halogen) or unsubstituted C1-6 alkyl, −C(=O)Raa, −CO2Raa, −C(=O)N(Rbb)2, or an oxygen protecting group, wherein Raa is hydrogen, substituted (e.g., substituted with one or more halogen) or unsubstituted C1-10 alkyl, or an oxygen protecting group when attached to an oxygen atom; and each Rbb is independently hydrogen, substituted (e.g., substituted with one or more halogen) or unsubstituted C1-10 alkyl, or a nitrogen protecting group. In certain embodiments, each oxygen atom substituent is independently substituted (e.g., substituted with one or more halogen) or unsubstituted C1-6 alkyl or an oxygen protecting group. [0088] In certain embodiments, the substituent present on an oxygen atom is an oxygen protecting group (also referred to herein as an “hydroxyl protecting group”). Oxygen protecting groups include
U1197.70243WO00 19/193 #13587456v2 −Raa, −N(Rbb)2, −C(=O)SRaa, −C(=O)Raa, −CO2Raa, −C(=O)N(Rbb)2, −C(=NRbb)Raa, −C(=NRbb)ORaa, −C(=NRbb)N(Rbb)2, −S(=O)Raa, −SO2Raa, −Si(Raa)3, −P(Rcc)2, −P(Rcc)3 +X−, −P(ORcc)2, −P(ORcc)3 +X−, −P(=O)(Raa)2, −P(=O)(ORcc)2, and −P(=O)(N(Rbb) 2)2, wherein X−, Raa, Rbb, and Rcc are as defined 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. Wuts, 3rd edition, John Wiley & Sons, 1999, incorporated herein by reference. [0089] A “counterion” or “anionic counterion” is a negatively charged group associated with a positively charged group in order to maintain electronic neutrality. An anionic counterion may be monovalent (e.g., including one formal negative charge). An anionic counterion may also be multivalent (e.g., including more than one formal negative charge), such as divalent or trivalent. Exemplary counterions include halide ions (e.g., F–, Cl–, Br–, I–), NO3–, ClO4–, OH–, H2PO4–, HCO3−, HSO4–, sulfonate ions (e.g., methansulfonate, trifluoromethanesulfonate, p–toluenesulfonate, benzenesulfonate, 10–camphor sulfonate, naphthalene–2–sulfonate, naphthalene–1–sulfonic acid–5– sulfonate, ethan–1–sulfonic acid–2–sulfonate, and the like), carboxylate ions (e.g., acetate, propanoate, benzoate, glycerate, lactate, tartrate, glycolate, gluconate, and the like), BF4−, PF4–, PF6–, AsF6–, SbF6–, B[3,5-(CF3)2C6H3]4]–, B(C6F5)4−, BPh4–, Al(OC(CF3)3)4–, and carborane anions (e.g., CB11H12– or (HCB11Me5Br6)–). Exemplary counterions which may be multivalent include CO32−, HPO42−, PO43−, B4O72−, SO42−, S2O32−, carboxylate anions (e.g., tartrate, citrate, fumarate, maleate, malate, malonate, gluconate, succinate, glutarate, adipate, pimelate, suberate, azelate, sebacate, salicylate, phthalates, aspartate, glutamate, and the like), and carboranes. [0090] A “leaving group” (LG) is an art-understood term referring to an atomic or molecular fragment that departs with a pair of electrons in heterolytic bond cleavage, wherein the molecular fragment is an anion or neutral molecule. As used herein, a leaving group can be an atom or a group capable of being displaced by a nucleophile. See e.g., Smith, March Advanced Organic Chemistry 6th ed. (501–502). In some embodiments, the leaving group is an internal leaving group such as an epoxide. [0091] Use of the phrase “at least one instance” refers to 1, 2, 3, 4, or more instances, but also encompasses a range, e.g., for example, from 1 to 4, from 1 to 3, from 1 to 2, from 2 to 4, from 2 to 3, or from 3 to 4 instances, inclusive. [0092] A “non-hydrogen group” refers to any group that is defined for a particular variable that is not hydrogen. [0093] The term “heteroatom” refers to an atom that is not hydrogen or carbon. In certain embodiments, the heteroatom is nitrogen. In certain embodiments, the heteroatom is oxygen. In certain embodiments, the heteroatom is sulfur. [0094] These and other exemplary substituents are described in more detail in the Detailed Description, Examples, and Claims. The present disclosure is not limited in any manner by the above exemplary listing of substituents.
U1197.70243WO00 20/193 #13587456v2 [0095] As used herein, the term “salt” refers to any and all salts, and encompasses pharmaceutically acceptable salts. Salts include ionic compounds that result from the neutralization reaction of an acid and a base. A salt is composed of one or more cations (positively charged ions) and one or more anions (negative ions) so that the salt is electrically neutral (without a net charge). Salts of the compounds of the present disclosure include those derived from inorganic and organic acids and bases. Examples of acid addition salts are salts of an amino group formed with inorganic acids, such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid, or with organic acids, such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid or by using other methods known in the art such as ion exchange. Other salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2–hydroxy–ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2–naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3–phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, hippurate, and the like. Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N+(C1–4 alkyl)4 salts. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Further salts include ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, lower alkyl sulfonate, and aryl sulfonate. [0096] The term “pharmaceutically acceptable salt” refers to those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response, and the like, and are commensurate with a reasonable benefit/risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, Berge et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19, incorporated herein by reference. Pharmaceutically acceptable salts of the compounds of the present disclosure include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable, nontoxic acid addition salts are salts of an amino group formed with inorganic acids, such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid or with organic acids, such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid or by using other methods known in the art such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2- hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate,
U1197.70243WO00 21/193 #13587456v2 methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and the like. Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium, and N+(C1-4 alkyl)4 salts. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, lower alkyl sulfonate, and aryl sulfonate. [0097] It is also to be understood that compounds that have the same molecular formula but differ in the nature or sequence of bonding of their atoms or the arrangement of their atoms in space are termed “isomers”. Isomers that differ in the arrangement of their atoms in space are termed “stereoisomers”. [0098] Stereoisomers that are not mirror images of one another are termed “diastereomers” and those that are non-superimposable mirror images of each other are termed “enantiomers”. When a compound has an asymmetric center, for example, it is bonded to four different groups, a pair of enantiomers is possible. An enantiomer can be characterized by the absolute configuration of its asymmetric center and is described by the R- and S-sequencing rules of Cahn and Prelog, or by the manner in which the molecule rotates the plane of polarized light and designated as dextrorotatory or levorotatory (i.e., as (+) or (−)-isomers respectively). A chiral compound can exist as either individual enantiomer or as a mixture thereof. A mixture containing equal proportions of the enantiomers is called a “racemic mixture”. [0099] The terms “composition” and “formulation” are used interchangeably. [0100] A “subject” to which administration is contemplated refers to a human (i.e., male or female of any age group, e.g., pediatric subject (e.g., infant, child, or adolescent) or adult subject (e.g., young adult, middle-aged adult, or senior adult)) or non-human animal. In certain embodiments, the non- human animal is a mammal (e.g., primate (e.g., cynomolgus monkey or rhesus monkey), commercially relevant mammal (e.g., cattle, pig, horse, sheep, goat, cat, or dog), or bird (e.g., commercially relevant bird, such as chicken, duck, goose, or turkey)). In certain embodiments, the non-human animal is a fish, reptile, or amphibian. The non-human animal may be a male or female at any stage of development. The non-human animal may be a transgenic animal or genetically engineered animal. The term “patient” refers to a human subject in need of treatment of a disease. [0101] The term “biological sample” refers to any sample including tissue samples (such as tissue sections and needle biopsies of a tissue); cell samples (e.g., cytological smears (such as Pap or blood smears) or samples of cells obtained by microdissection); samples of whole organisms (such as samples of yeasts or bacteria); or cell fractions, fragments or organelles (such as obtained by lysing cells and separating the components thereof by centrifugation or otherwise). Other examples of biological samples include blood, serum, urine, semen, fecal matter, cerebrospinal fluid, interstitial
U1197.70243WO00 22/193 #13587456v2 fluid, mucous, tears, sweat, pus, biopsied tissue (e.g., obtained by a surgical biopsy or needle biopsy), nipple aspirates, milk, vaginal fluid, saliva, swabs (such as buccal swabs), or any material containing biomolecules that is derived from a first biological sample. [0102] The term “target tissue” refers to any biological tissue of a subject (including a group of cells, a body part, or an organ) or a part thereof, including blood and/or lymph vessels, which is the object to which a compound, particle, and/or composition of the present disclosure is delivered. A target tissue may be an abnormal or unhealthy tissue, which may need to be treated. A target tissue may also be a normal or healthy tissue that is under a higher than normal risk of becoming abnormal or unhealthy, which may need to be prevented. In certain embodiments, the target tissue is the liver. In certain embodiments, the target tissue is the lung. A “non-target tissue” is any biological tissue of a subject (including a group of cells, a body part, or an organ) or a part thereof, including blood and/or lymph vessels, which is not a target tissue. [0103] The term “administer,” “administering,” or “administration” refers to implanting, absorbing, ingesting, injecting, inhaling, or otherwise introducing a compound described herein, or a composition thereof, in or on a subject. [0104] The terms “treatment,” “treat,” and “treating” refer to reversing, alleviating, delaying the onset of, or inhibiting the progress of a disease described herein. In some embodiments, treatment may be administered after one or more signs or symptoms of the disease have developed or have been observed. In other embodiments, treatment may be administered in the absence of signs or symptoms of the disease. For example, treatment may be administered to a susceptible subject prior to the onset of symptoms (e.g., in light of a history of symptoms and/or in light of exposure to a pathogen). Treatment may also be continued after symptoms have resolved, for example, to delay or prevent recurrence. [0105] The terms “condition,” “disease,” and “disorder” are used interchangeably. [0106] An “effective amount” of a compound described herein refers to an amount sufficient to elicit the desired biological response. An effective amount of a compound described herein may vary depending on such factors as the desired biological endpoint, severity of side effects, disease, or disorder, the identity, pharmacokinetics, and pharmacodynamics of the particular compound, the condition being treated, the mode, route, and desired or required frequency of administration, the species, age and health or general condition of the subject. In certain embodiments, an effective amount is a therapeutically effective amount. In certain embodiments, an effective amount is a prophylactic treatment. In certain embodiments, an effective amount is the amount of a compound described herein in a single dose. In certain embodiments, an effective amount is the combined amounts of a compound described herein in multiple doses. In certain embodiments, the desired dosage is delivered three times a day, two times a day, once a day, every other day, every third day, every week, every two weeks, every three weeks, or every four weeks. In certain embodiments, the
U1197.70243WO00 23/193 #13587456v2 desired dosage is delivered using multiple administrations (e.g., two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, or more administrations). [0107] In certain embodiments, an effective amount of a compound for administration one or more times a day to a 70 kg adult human comprises about 0.0001 mg to about 3000 mg, about 0.0001 mg to about 2000 mg, about 0.0001 mg to about 1000 mg, about 0.001 mg to about 1000 mg, about 0.01 mg to about 1000 mg, about 0.1 mg to about 1000 mg, about 1 mg to about 1000 mg, about 1 mg to about 100 mg, about 10 mg to about 1000 mg, or about 100 mg to about 1000 mg, of a compound per unit dosage form. [0108] In certain embodiments, the compounds of the present disclosure are administered orally or parenterally at dosage levels sufficient to deliver from about 0.001 mg/kg to about 100 mg/kg, from about 0.01 mg/kg to about 50 mg/kg, preferably from about 0.1 mg/kg to about 40 mg/kg, preferably from about 0.5 mg/kg to about 30 mg/kg, from about 0.01 mg/kg to about 10 mg/kg, from about 0.1 mg/kg to about 10 mg/kg, and more preferably from about 1 mg/kg to about 25 mg/kg, of subject body weight per day, one or more times a day, to obtain the desired therapeutic effect. [0109] It will be appreciated that dose ranges as described herein provide guidance for the administration of provided pharmaceutical compositions to an adult. The amount to be administered to, for example, a child or an adolescent can be determined by a medical practitioner or person skilled in the art and can be lower or the same as that administered to an adult. [0110] A “therapeutically effective amount” of a compound described herein is an amount sufficient to provide a therapeutic benefit in the treatment of a condition or to delay or minimize one or more symptoms associated with the condition. A therapeutically effective amount of a compound means an amount of therapeutic agent, alone or in combination with other therapies, which provides a therapeutic benefit in the treatment of the condition. The term “therapeutically effective amount” can encompass an amount that improves overall therapy, reduces or avoids symptoms, signs, or causes of the condition, and/or enhances the therapeutic efficacy of another therapeutic agent. In certain embodiments, a therapeutically effective amount is an amount sufficient for modulating a target protein in a subject or a cell, tissue, or biological sample. In certain embodiments, a therapeutically effective amount is an amount sufficient for treating cancer (e.g., a solid tumor (e.g., brain tumor (e.g., glioblastoma), pancreatic cancer, ovarian cancer)). In certain embodiments, a therapeutically effective amount is an amount sufficient for modulating a target protein in a subject or a cell, tissue, or biological sample and treating cancer (e.g., a solid tumor (e.g., brain tumor (e.g., glioblastoma), pancreatic cancer, ovarian cancer)). [0111] A “prophylactically effective amount” of a compound described herein is an amount sufficient to prevent a condition, or one or more symptoms associated with the condition or prevent its recurrence. A prophylactically effective amount of a compound means an amount of a therapeutic agent, alone or in combination with other agents, which provides a prophylactic benefit in the prevention of the condition. The term “prophylactically effective amount” can encompass an amount
U1197.70243WO00 24/193 #13587456v2 that improves overall prophylaxis or enhances the prophylactic efficacy of another prophylactic agent. In certain embodiments, a prophylactically effective amount is an amount sufficient for modulating a target protein in a subject or a cell, tissue, or biological sample. In certain embodiments, a prophylactically effective amount is an amount sufficient for treating cancer (e.g., a solid tumor (e.g., brain tumor (e.g., glioblastoma), pancreatic cancer, ovarian cancer)). In certain embodiments, a prophylactically effective amount is an amount sufficient for modulating a target protein in a subject or a cell, tissue, or biological sample and treating cancer (e.g., a solid tumor (e.g., brain tumor (e.g., glioblastoma), pancreatic cancer, ovarian cancer)). [0112] The term “prevent,” “preventing,” or “prevention” refers to a prophylactic treatment of a subject who is not and was not with a disease but is at risk of developing the disease or who was with a disease, is not with the disease, but is at risk of regression of the disease. In certain embodiments, the subject is at a higher risk of developing the disease or at a higher risk of regression of the disease than an average healthy member of a population. [0113] The term “inhibit” or “inhibition” in the context of enzymes, for example, in the context of STAT3, IDO, NOS, arginase, PD-L1, protein kinase B, or CD73, refers to a reduction in the activity of the enzyme. In some embodiments, the term refers to a reduction of the level of enzyme activity, e.g., STAT3, IDO, NOS, arginase, PD-L1, protein kinase B, or CD73 activity, to a level that is statistically significantly lower than an initial level, which may, for example, be a baseline level of enzyme activity. In some embodiments, the term refers to a reduction of the level of enzyme activity, e.g., STAT3, IDO, NOS, arginase, PD-L1, protein kinase B, or CD73 activity, to a level that is less than 75%, less than 50%, less than 40%, less than 30%, less than 25%, less than 20%, less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, less than 0.01%, less than 0.001%, or less than 0.0001% of an initial level, which may, for example, be a baseline level of enzyme activity. [0114] The term “agonist” or “agonism” in the context of enzymes, for example, in the context of TLR7 or STING, refers to the activation of a receptor to produce an enzyme response. In some embodiments, the term refers to an increase of the level of enzyme response, e.g., TLR7 or STING response, to a level that is statistically significantly higher than an initial level, which may, for example, be a baseline level of enzyme response. In some embodiments, the term refers to an increase of the level of enzyme response, e.g., TLR7 or STING response, to a level that is greater than 100%, greater than 125%, greater than 150%, greater than 175%, greater than 200%, greater than 300%, greater than 400%, greater than 500%, greater than 600%, greater than 700%, greater than 800%, greater than 900%, or greater than 1000% of an initial level, which may, for example, be a baseline level of enzyme response. [0115] The term “antagonist” or “antagonism” in the context of enzymes, for example, refers to blocking activation of a receptor to prevent an enzyme response. In some embodiments, the term refers to a reduction of the level of enzyme response, to a level that is statistically significantly lower
U1197.70243WO00 25/193 #13587456v2 than an initial level, which may, for example, be a baseline level of enzyme response. In some embodiments, the term refers to a reduction of the level of enzyme response, to a level that is less than 75%, less than 50%, less than 40%, less than 30%, less than 25%, less than 20%, less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, less than 0.01%, less than 0.001%, or less than 0.0001% of an initial level, which may, for example, be a baseline level of enzyme response. [0116] A “proliferative disease” refers to a disease that occurs due to abnormal growth or extension by the multiplication of cells (Walker, Cambridge Dictionary of Biology; Cambridge University Press: Cambridge, UK, 1990). A proliferative disease may be associated with: 1) the pathological proliferation of normally quiescent cells; 2) the pathological migration of cells from their normal location (e.g., metastasis of neoplastic cells); 3) the pathological expression of proteolytic enzymes such as the matrix metalloproteinases (e.g., collagenases, gelatinases, and elastases); or 4) the pathological angiogenesis as in proliferative retinopathy and tumor metastasis. Exemplary proliferative diseases include cancers (i.e., “malignant neoplasms”), benign neoplasms, angiogenesis, inflammatory diseases, and autoimmune diseases. [0117] The term “angiogenesis” refers to the physiological process through which new blood vessels form from pre-existing vessels. Angiogenesis is distinct from vasculogenesis, which is the de novo formation of endothelial cells from mesoderm cell precursors. The first vessels in a developing embryo form through vasculogenesis, after which angiogenesis is responsible for most blood vessel growth during normal or abnormal development. Angiogenesis is a vital process in growth and development, as well as in wound healing and in the formation of granulation tissue. However, angiogenesis is also a fundamental step in the transition of tumors from a benign state to a malignant one, leading to the use of angiogenesis inhibitors in the treatment of cancer. Angiogenesis may be chemically stimulated by angiogenic proteins, such as growth factors (e.g., VEGF). “Pathological angiogenesis” refers to abnormal (e.g., excessive or insufficient) angiogenesis that amounts to and/or is associated with a disease. [0118] The terms “neoplasm” and “tumor” are used herein interchangeably and refer to an abnormal mass of tissue wherein the growth of the mass surpasses and is not coordinated with the growth of a normal tissue. A neoplasm or tumor may be “benign” or “malignant,” depending on the following characteristics: degree of cellular differentiation (including morphology and functionality), rate of growth, local invasion, and metastasis. A “benign neoplasm” is generally well differentiated, has characteristically slower growth than a malignant neoplasm, and remains localized to the site of origin. In addition, a benign neoplasm does not have the capacity to infiltrate, invade, or metastasize to distant sites. Exemplary benign neoplasms include, but are not limited to, lipoma, chondroma, adenomas, acrochordon, senile angiomas, seborrheic keratoses, lentigos, and sebaceous hyperplasias. In some cases, certain “benign” tumors may later give rise to malignant neoplasms, which may result from additional genetic changes in a subpopulation of the tumor’s neoplastic cells, and these tumors
U1197.70243WO00 26/193 #13587456v2 are referred to as “pre-malignant neoplasms.” An exemplary pre-malignant neoplasm is a teratoma. In contrast, a “malignant neoplasm” is generally poorly differentiated (anaplasia) and has characteristically rapid growth accompanied by progressive infiltration, invasion, and destruction of the surrounding tissue. Furthermore, a malignant neoplasm generally has the capacity to metastasize to distant sites. The term “metastasis,” “metastatic,” or “metastasize” refers to the spread or migration of cancerous cells from a primary or original tumor to another organ or tissue and is typically identifiable by the presence of a “secondary tumor” or “secondary cell mass” of the tissue type of the primary or original tumor and not of that of the organ or tissue in which the secondary (metastatic) tumor is located. For example, a prostate cancer that has migrated to bone is said to be metastasized prostate cancer and includes cancerous prostate cancer cells growing in bone tissue. [0119] The term “cancer” refers to a class of diseases characterized by the development of abnormal cells that proliferate uncontrollably and have the ability to infiltrate and destroy normal body tissues. See e.g., Stedman’s Medical Dictionary, 25th ed.; Hensyl ed.; Williams & Wilkins: Philadelphia, 1990. Exemplary cancers include, but are not limited to, acoustic neuroma; adenocarcinoma; adrenal gland cancer; anal cancer; angiosarcoma (e.g., lymphangiosarcoma, lymphangioendotheliosarcoma, hemangiosarcoma); appendix cancer; benign monoclonal gammopathy; biliary cancer (e.g., cholangiocarcinoma); bladder cancer; breast cancer (e.g., adenocarcinoma of the breast, papillary carcinoma of the breast, mammary cancer, medullary carcinoma of the breast); brain cancer (e.g., meningioma, glioblastomas, glioma (e.g., astrocytoma, oligodendroglioma), medulloblastoma); bronchus cancer; carcinoid tumor; cervical cancer (e.g., cervical adenocarcinoma); choriocarcinoma; chordoma; craniopharyngioma; colorectal cancer (e.g., colon cancer, rectal cancer, colorectal adenocarcinoma); connective tissue cancer; epithelial carcinoma; ependymoma; endotheliosarcoma (e.g., Kaposi’s sarcoma, multiple idiopathic hemorrhagic sarcoma); endometrial cancer (e.g., uterine cancer, uterine sarcoma); esophageal cancer (e.g., adenocarcinoma of the esophagus, Barrett’s adenocarcinoma); Ewing’s sarcoma; ocular cancer (e.g., intraocular melanoma, retinoblastoma); familiar hypereosinophilia; gall bladder cancer; gastric cancer (e.g., stomach adenocarcinoma); gastrointestinal stromal tumor (GIST); germ cell cancer; head and neck cancer (e.g., head and neck squamous cell carcinoma, oral cancer (e.g., oral squamous cell carcinoma), throat cancer (e.g., laryngeal cancer, pharyngeal cancer, nasopharyngeal cancer, oropharyngeal cancer)); hematopoietic cancers (e.g., leukemia such as acute lymphocytic leukemia (ALL) (e.g., B-cell ALL, T-cell ALL), acute myelocytic leukemia (AML) (e.g., B-cell AML, T-cell AML), chronic myelocytic leukemia (CML) (e.g., B-cell CML, T-cell CML), and chronic lymphocytic leukemia (CLL) (e.g., B-cell CLL, T-cell CLL)); lymphoma such as Hodgkin lymphoma (HL) (e.g., B-cell HL, T-cell HL) and non- Hodgkin lymphoma (NHL) (e.g., B-cell NHL such as diffuse large cell lymphoma (DLCL) (e.g., diffuse large B-cell lymphoma), follicular lymphoma, chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL), mantle cell lymphoma (MCL), marginal zone B-cell lymphomas (e.g., mucosa-associated lymphoid tissue (MALT) lymphomas, nodal marginal zone B-cell
U1197.70243WO00 27/193 #13587456v2 lymphoma, splenic marginal zone B-cell lymphoma), primary mediastinal B-cell lymphoma, Burkitt lymphoma, lymphoplasmacytic lymphoma (i.e., Waldenström’s macroglobulinemia), hairy cell leukemia (HCL), immunoblastic large cell lymphoma, precursor B-lymphoblastic lymphoma and primary central nervous system (CNS) lymphoma; and T-cell NHL such as precursor T-lymphoblastic lymphoma/leukemia, peripheral T-cell lymphoma (PTCL) (e.g., cutaneous T-cell lymphoma (CTCL) (e.g., mycosis fungoides, Sezary syndrome), angioimmunoblastic T-cell lymphoma, extranodal natural killer T-cell lymphoma, enteropathy type T-cell lymphoma, subcutaneous panniculitis-like T- cell lymphoma, and anaplastic large cell lymphoma); a mixture of one or more leukemia/lymphoma as described above; and multiple myeloma (MM)), heavy chain disease (e.g., alpha chain disease, gamma chain disease, mu chain disease); hemangioblastoma; hypopharynx cancer; inflammatory myofibroblastic tumors; immunocytic amyloidosis; kidney cancer (e.g., nephroblastoma a.k.a. Wilms’ tumor, renal cell carcinoma); liver cancer (e.g., hepatocellular cancer (HCC), malignant hepatoma); lung cancer (e.g., bronchogenic carcinoma, small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), adenocarcinoma of the lung); leiomyosarcoma (LMS); mastocytosis (e.g., systemic mastocytosis); muscle cancer; myelodysplastic syndrome (MDS); mesothelioma; myeloproliferative disorder (MPD) (e.g., polycythemia vera (PV), essential thrombocytosis (ET), agnogenic myeloid metaplasia (AMM) a.k.a. myelofibrosis (MF), chronic idiopathic myelofibrosis, chronic myelocytic leukemia (CML), chronic neutrophilic leukemia (CNL), hypereosinophilic syndrome (HES)); neuroblastoma; neurofibroma (e.g., neurofibromatosis (NF) type 1 or type 2, schwannomatosis); neuroendocrine cancer (e.g., gastroenteropancreatic neuroendoctrine tumor (GEP-NET), carcinoid tumor); osteosarcoma (e.g.,bone cancer); ovarian cancer (e.g., cystadenocarcinoma, ovarian embryonal carcinoma, ovarian adenocarcinoma); papillary adenocarcinoma; pancreatic cancer (e.g., pancreatic andenocarcinoma, intraductal papillary mucinous neoplasm (IPMN), Islet cell tumors); penile cancer (e.g., Paget’s disease of the penis and scrotum); pinealoma; primitive neuroectodermal tumor (PNT); plasma cell neoplasia; paraneoplastic syndromes; intraepithelial neoplasms; prostate cancer (e.g., prostate adenocarcinoma); rectal cancer; rhabdomyosarcoma; salivary gland cancer; skin cancer (e.g., squamous cell carcinoma (SCC), keratoacanthoma (KA), melanoma, basal cell carcinoma (BCC)); small bowel cancer (e.g., appendix cancer); soft tissue sarcoma (e.g., malignant fibrous histiocytoma (MFH), liposarcoma, malignant peripheral nerve sheath tumor (MPNST), chondrosarcoma, fibrosarcoma, myxosarcoma); sebaceous gland carcinoma; small intestine cancer; sweat gland carcinoma; synovioma; testicular cancer (e.g., seminoma, testicular embryonal carcinoma); thyroid cancer (e.g., papillary carcinoma of the thyroid, papillary thyroid carcinoma (PTC), medullary thyroid cancer); urethral cancer; vaginal cancer; and vulvar cancer (e.g., Paget’s disease of the vulva). [0120] The terms “inflammatory disease” and “inflammatory condition” are used interchangeably herein, and refer to a disease or condition caused by, resulting from, or resulting in inflammation. Inflammatory diseases and conditions include those diseases, disorders or conditions that are
U1197.70243WO00 28/193 #13587456v2 characterized by signs of pain (dolor, from the generation of noxious substances and the stimulation of nerves), heat (calor, from vasodilatation), redness (rubor, from vasodilatation and increased blood flow), swelling (tumor, from excessive inflow or restricted outflow of fluid), and/or loss of function (functio laesa, which can be partial or complete, temporary or permanent. Inflammation takes on many forms and includes, but is not limited to, acute, adhesive, atrophic, catarrhal, chronic, cirrhotic, diffuse, disseminated, exudative, fibrinous, fibrosing, focal, granulomatous, hyperplastic, hypertrophic, interstitial, metastatic, necrotic, obliterative, parenchymatous, plastic, productive, proliferous, pseudomembranous, purulent, sclerosing, seroplastic, serous, simple, specific, subacute, suppurative, toxic, traumatic, and/or ulcerative inflammation. The term “inflammatory disease” may also refer to a dysregulated inflammatory reaction that causes an exaggerated response by macrophages, granulocytes, and/or T-lymphocytes leading to abnormal tissue damage and/or cell death. An inflammatory disease can be either an acute or chronic inflammatory condition and can result from infections or non-infectious causes. Inflammatory diseases include, without limitation, atherosclerosis, arteriosclerosis, autoimmune disorders, multiple sclerosis, systemic lupus erythematosus, polymyalgia rheumatica (PMR), gouty arthritis, degenerative arthritis, tendonitis, bursitis, psoriasis, cystic fibrosis, arthrosteitis, rheumatoid arthritis, inflammatory arthritis, Sjogren’s syndrome, giant cell arteritis, progressive systemic sclerosis (scleroderma), ankylosing spondylitis, polymyositis, dermatomyositis, pemphigus, pemphigoid, diabetes (e.g., Type I), myasthenia gravis, Hashimoto’s thyroiditis, Graves’ disease, Goodpasture’s disease, mixed connective tissue disease, sclerosing cholangitis, inflammatory bowel disease, Crohn’s disease, ulcerative colitis, pernicious anemia, inflammatory dermatoses, usual interstitial pneumonitis (UIP), asbestosis, silicosis, bronchiectasis, berylliosis, talcosis, pneumoconiosis, sarcoidosis, desquamative interstitial pneumonia, lymphoid interstitial pneumonia, giant cell interstitial pneumonia, cellular interstitial pneumonia, extrinsic allergic alveolitis, Wegener’s granulomatosis and related forms of angiitis (temporal arteritis and polyarteritis nodosa), inflammatory dermatoses, hepatitis, delayed-type hypersensitivity reactions (e.g., poison ivy dermatitis), pneumonia, respiratory tract inflammation, Adult Respiratory Distress Syndrome (ARDS), encephalitis, immediate hypersensitivity reactions, asthma, hayfever, allergies, acute anaphylaxis, rheumatic fever, glomerulonephritis, pyelonephritis, cellulitis, cystitis, chronic cholecystitis, ischemia (ischemic injury), reperfusion injury, allograft rejection, host-versus-graft rejection, appendicitis, arteritis, blepharitis, bronchiolitis, bronchitis, cervicitis, cholangitis, chorioamnionitis, conjunctivitis, dacryoadenitis, dermatomyositis, endocarditis, endometritis, enteritis, enterocolitis, epicondylitis, epididymitis, fasciitis, fibrositis, gastritis, gastroenteritis, gingivitis, ileitis, iritis, laryngitis, myelitis, myocarditis, nephritis, omphalitis, oophoritis, orchitis, osteitis, otitis, pancreatitis, parotitis, pericarditis, pharyngitis, pleuritis, phlebitis, pneumonitis, proctitis, prostatitis, rhinitis, salpingitis, sinusitis, stomatitis, synovitis, testitis, tonsillitis, urethritis, urocystitis, uveitis, vaginitis, vasculitis, vulvitis, vulvovaginitis, angitis, chronic bronchitis, osteomyelitis, optic neuritis, temporal arteritis, transverse myelitis, necrotizing fasciitis, and
U1197.70243WO00 29/193 #13587456v2 necrotizing enterocolitis. An ocular inflammatory disease includes, but is not limited to, post-surgical inflammation. [0121] Additional exemplary inflammatory conditions include, but are not limited to, inflammation associated with acne, anemia (e.g., aplastic anemia, hemolytic autoimmune anemia), asthma, arteritis (e.g., polyarteritis, temporal arteritis, periarteritis nodosa, Takayasu’s arteritis), arthritis (e.g., crystalline arthritis, osteoarthritis, psoriatic arthritis, gouty arthritis, reactive arthritis, rheumatoid arthritis and Reiter’s arthritis), ankylosing spondylitis, amylosis, amyotrophic lateral sclerosis, autoimmune diseases, allergies or allergic reactions, atherosclerosis, bronchitis, bursitis, chronic prostatitis, conjunctivitis, Chagas disease, chronic obstructive pulmonary disease, cermatomyositis, diverticulitis, diabetes (e.g., type I diabetes mellitus, Type II diabetes mellitus), a skin condition (e.g., psoriasis, eczema, burns, dermatitis, pruritus (itch)), endometriosis, Guillain-Barre syndrome, infection, ischemic heart disease, Kawasaki disease, glomerulonephritis, gingivitis, hypersensitivity, headaches (e.g., migraine headaches, tension headaches), ileus (e.g., postoperative ileus and ileus during sepsis), idiopathic thrombocytopenic purpura, interstitial cystitis (painful bladder syndrome), gastrointestinal disorder (e.g., selected from peptic ulcers, regional enteritis, diverticulitis, gastrointestinal bleeding, eosinophilic gastrointestinal disorders (e.g., eosinophilic esophagitis, eosinophilic gastritis, eosinophilic gastroenteritis, eosinophilic colitis), gastritis, diarrhea, gastroesophageal reflux disease (GORD, or its synonym GERD), inflammatory bowel disease (IBD) (e.g., Crohn’s disease, ulcerative colitis, collagenous colitis, lymphocytic colitis, ischemic colitis, diversion colitis, Behcet’s syndrome, indeterminate colitis) and inflammatory bowel syndrome (IBS)), lupus, multiple sclerosis, morphea, myasthenia gravis, myocardial ischemia, nephrotic syndrome, pemphigus vulgaris, pernicious anemia, peptic ulcers, polymyositis, primary biliary cirrhosis, neuroinflammation associated with brain disorders (e.g., Parkinson’s disease, Huntington’s disease, and Alzheimer’s disease), prostatitis, chronic inflammation associated with cranial radiation injury, pelvic inflammatory disease, reperfusion injury, regional enteritis, rheumatic fever, systemic lupus erythematosus, scleroderma, sarcoidosis, spondyloarthopathies, Sjogren’s syndrome, thyroiditis, transplantation rejection, tendonitis, trauma or injury (e.g., frostbite, chemical irritants, toxins, scarring, burns, physical injury), vasculitis, vitiligo and Wegener’s granulomatosis. In certain embodiments, the inflammatory disorder is selected from arthritis (e.g., rheumatoid arthritis), inflammatory bowel disease, inflammatory bowel syndrome, asthma, psoriasis, endometriosis, interstitial cystitis and prostatitis. In certain embodiments, the inflammatory condition is an acute inflammatory condition (e.g., for example, inflammation resulting from infection). In certain embodiments, the inflammatory condition is a chronic inflammatory condition (e.g., conditions resulting from asthma, arthritis and inflammatory bowel disease). The compounds may also be useful in treating inflammation associated with trauma and non-inflammatory myalgia. The compounds disclosed herein may also be useful in treating inflammation associated with cancer.
U1197.70243WO00 30/193 #13587456v2 [0122] An “autoimmune disease” refers to a disease arising from an inappropriate immune response of the body of a subject against substances and tissues normally present in the body. In other words, the immune system mistakes some part of the body as a pathogen and attacks its own cells. This may be restricted to certain organs (e.g., in autoimmune thyroiditis) or involve a particular tissue in different places (e.g., Goodpasture’s disease which may affect the basement membrane in both the lung and kidney). The treatment of autoimmune diseases is typically with immunosuppression, e.g., medications which decrease the immune response. Exemplary autoimmune diseases include, but are not limited to, glomerulonephritis, Goodpasture’s syndrome, necrotizing vasculitis, lymphadenitis, peri-arteritis nodosa, systemic lupus erythematosis, rheumatoid arthritis, psoriatic arthritis, , psoriasis, ulcerative colitis, systemic sclerosis, dermatomyositis/polymyositis, anti-phospholipid antibody syndrome, scleroderma, pemphigus vulgaris, ANCA-associated vasculitis (e.g., Wegener’s granulomatosis, microscopic polyangiitis), uveitis, Sjogren’s syndrome, Crohn’s disease, Reiter’s syndrome, ankylosing spondylitis, Lyme disease, Guillain-Barré syndrome, Hashimoto’s thyroiditis, and cardiomyopathy. [0123] The term “liver disease” or “hepatic disease” refers to damage to or a disease of the liver. Non-limiting examples of liver disease include intrahepatic cholestasis (e.g., alagille syndrome, biliary liver cirrhosis), fatty liver (e.g., alcoholic fatty liver, Reye’s syndrome), hepatic vein thrombosis, hepatolenticular degeneration (i.e., Wilson’s disease), hepatomegaly, liver abscess (e.g., amebic liver abscess), liver cirrhosis (e.g., alcoholic, biliary, and experimental liver cirrhosis), alcoholic liver diseases (e.g., fatty liver, hepatitis, cirrhosis), parasitic liver disease (e.g., hepatic echinococcosis, fascioliasis, amebic liver abscess), jaundice (e.g., hemolytic, hepatocellular, cholestatic jaundice), cholestasis, portal hypertension, liver enlargement, ascites, hepatitis (e.g., alcoholic hepatitis, animal hepatitis, chronic hepatitis (e.g., autoimmune, hepatitis B, hepatitis C, hepatitis D, drug induced chronic hepatitis), toxic hepatitis, viral human hepatitis (e.g., hepatitis A, hepatitis B, hepatitis C, hepatitis D, hepatitis E), granulomatous hepatitis, secondary biliary cirrhosis, hepatic encephalopathy, varices, primary biliary cirrhosis, primary sclerosing cholangitis, hepatocellular adenoma, hemangiomas, bile stones, liver failure (e.g., hepatic encephalopathy, acute liver failure), angiomyolipoma, calcified liver metastases, cystic liver metastases, fibrolamellar hepatocarcinoma, hepatic adenoma, hepatoma, hepatic cysts (e.g., Simple cysts, Polycystic liver disease, hepatobiliary cystadenoma, choledochal cyst), mesenchymal tumors (mesenchymal hamartoma, infantile hemangioendothelioma, hemangioma, peliosis hepatis, lipomas, inflammatory pseudotumor), epithelial tumors (e.g., bile duct hamartoma, bile duct adenoma), focal nodular hyperplasia, nodular regenerative hyperplasia, hepatoblastoma, hepatocellular carcinoma, cholangiocarcinoma, cystadenocarcinoma, tumors of blood vessels, angiosarcoma, Karposi’s sarcoma, hemangioendothelioma, embryonal sarcoma, fibrosarcoma, leiomyosarcoma, rhabdomyosarcoma, carcinosarcoma, teratoma, carcinoid, squamous carcinoma, primary lymphoma, peliosis hepatis,
U1197.70243WO00 31/193 #13587456v2 erythrohepatic porphyria, hepatic porphyria (e.g., acute intermittent porphyria, porphyria cutanea tarda), and Zellweger syndrome. [0124] Immune disorders, such as auto-immune disorders, include, but are not limited to, arthritis (including rheumatoid arthritis, spondyloarthopathies, gouty arthritis, degenerative joint diseases such as osteoarthritis, systemic lupus erythematosus, Sjogren’s syndrome, ankylosing spondylitis, undifferentiated spondylitis, Behcet’s disease, haemolytic autoimmune anaemias, multiple sclerosis, amyotrophic lateral sclerosis, amylosis, acute painful shoulder, psoriatic, and juvenile arthritis), asthma, atherosclerosis, osteoporosis, bronchitis, tendonitis, bursitis, skin condition (e.g., psoriasis, eczema, burns, dermatitis, pruritus (itch)), enuresis, eosinophilic disease, gastrointestinal disorder (e.g., selected from peptic ulcers, regional enteritis, diverticulitis, gastrointestinal bleeding, eosinophilic gastrointestinal disorders (e.g., eosinophilic esophagitis, eosinophilic gastritis, eosinophilic gastroenteritis, eosinophilic colitis), gastritis, diarrhea, gastroesophageal reflux disease (GORD, or its synonym GERD), inflammatory bowel disease (IBD) (e.g., Crohn’s disease, ulcerative colitis, collagenous colitis, lymphocytic colitis, ischaemic colitis, diversion colitis, Behcet’s syndrome, indeterminate colitis) and inflammatory bowel syndrome (IBS)), and disorders ameliorated by a gastroprokinetic agent (e.g., ileus, postoperative ileus and ileus during sepsis; gastroesophageal reflux disease (GORD, or its synonym GERD); eosinophilic esophagitis, gastroparesis such as diabetic gastroparesis; food intolerances and food allergies and other functional bowel disorders, such as non-ulcerative dyspepsia (NUD) and non-cardiac chest pain (NCCP, including costo-chondritis)). [0125] A “microbial infection” refers to an infection with a microorganism, such as a fungus, bacteria or virus. In certain embodiments, the microbial infection is an infection with a fungus, i.e., a fungal infection. In certain embodiments, the microbial infection is an infection with a virus, i.e., a viral infection. In certain embodiments, the microbial infection is an infection with a bacteria, i.e., a bacterial infection. Various microbial infections include, but are not limited to, skin infections, GI infections, urinary tract infections, genito-urinary infections, sepsis, blood infections, and systemic infections. [0126] The term “particle” refers to a small object, fragment, or piece of a substance that may be a single element, inorganic material, organic material, or mixture thereof. Examples of particles include polymeric particles, single-emulsion particles, double-emulsion particles, coacervates, liposomes, microparticles, nanoparticles, macroscopic particles, pellets, crystals, aggregates, composites, pulverized, milled or otherwise disrupted matrices, and cross-linked protein or polysaccharide particles, each of which have an average characteristic dimension of about less than about 1 mm and at least 1 nm, where the characteristic dimension, or “critical dimension,” of the particle is the smallest cross-sectional dimension of the particle. A particle may be composed of a single substance or multiple substances. In certain embodiments, the particle is not a viral particle. In other embodiments, the particle is not a liposome. In certain embodiments, the particle is not a micelle. In
U1197.70243WO00 32/193 #13587456v2 certain embodiments, the particle is substantially solid throughout. In certain embodiments, the particle is a nanoparticle. In certain embodiments, the particle is a microparticle. [0127] The term “nanoparticle” refers to a particle having an average (e.g., mean) dimension (e.g., diameter) of between about 1 nanometer (nm) and about 1 micrometer (µm) (e.g., between about 1 nm and about 300 nm, between about 1 nm and about 100 nm, between about 1 nm and about 30 nm, between about 1 nm and about 10 nm, or between about 1 nm and about 3 nm), inclusive. [0128] The term “microparticle” refers to a particle having an average (e.g., mean) dimension (e.g., diameter) of between about 1 micrometer (µm) and about 1 millimeter (mm) (e.g., between about 1 µm and about 100 µm, between about 1 µm and about 30 µm, between about 1 µm and about 10 µm, or between about 1 µm and about 3 µm), inclusive. [0129] The “hydrodynamic diameter” of a particle refers to the diameter of a solid sphere that would exhibit the same hydrodynamic friction as the particle (e.g., the diameter of a solid sphere that diffuses at the same rate as the particle). Hydrodynamic diameter can be measured through various techniques including dynamic light scattering (DLS) and nanoparticle tracking analysis (NTA). [0130] The terms “polydispersity index” or “PDI” refer to the degree of non-uniformity of a size distribution of particles (e.g., the broadness of a molecular weight distribution). PDI can be measured through various techniques including dynamic light scattering (DLS). [0131] The term “zeta potential” refers to the potential difference between the surface of a particle and the surrounding liquid the particles are dispersed in (e.g., the surface charge of nanoparticles in solution). Zeta potential can be measured through various techniques including electrophoretic light scattering and electroacoustic phenomenon. [0132] The term “pKa” refers to the negative decadic logarithm of the ionization constant (Ka) of an acid; equal to the pH value at which equal concentrations of the acid and conjugate base forms of a substance (often a buffer) are present. [0133] The term “about X,” where X is a number or percentage, refers to a number or percentage that is between 99.5% and 100.5%, between 99% and 101%, between 98% and 102%, between 97% and 103%, between 96% and 104%, between 95% and 105%, between 92% and 108%, or between 90% and 110%, inclusive, of X. [0134] The term “dendrimer” refers to a molecular architecture with an interior core, interior layers (or “generations”) of repeating units which are attached to and extend from this interior core, each layer having one or more branching points, and the outermost generation having terminal functional groups on an exterior surface. Examples of dendrimers include, but are not limited to, poly(amidoamine) (PAMAM), polyester, polylysine, and poly(propylene imine) (PPI). In some embodiments, terminal functional groups of a dendrimer include one or more hydroxyl groups, one or more amine groups, and/or one or more carboxyl groups. [0135] The term “PAMAM dendrimer” means poly(amidoamine) dendrimer, which may contain different cores, with amidoamine building blocks. The method for making them is known to those of
U1197.70243WO00 33/193 #13587456v2 skill in the art and generally, involves a two-step iterative reaction sequence that produces concentric shells (generations) of dendritic 0-alanine units around a central initiator core. This PAMAM core- shell architecture grows linearly in diameter as a function of added shells (generations). Meanwhile, the surface groups amplify exponentially at each generation according to dendritic-branching mathematics. PAMAM dendrimers can have carboxylic acid, amine and/or hydroxyl terminations and can be any generation of dendrimers including, but not limited to, generation 1 PAMAM dendrimers, generation 2 PAMAM dendrimers, generation 3 PAMAM dendrimers, generation 4 PAMAM dendrimers, generation 5 PAMAM dendrimers, generation 6 PAMAM dendrimers, generation 7 PAMAM dendrimers, generation 8 PAMAM dendrimers, generation 9 PAMAM dendrimers, or generation 10 PAMAM dendrimers. In some embodiments, the number of terminal sites on a dendrimer can depend on the particular dendrimeric scaffold and its generation. For example, in some embodiments, a dendrimer is based on a generation 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 PAMAM dendrimeric scaffold, which have 4, 8, 16, 32, 64, 128, 256, 512, 1024, 2048, and 4096 terminal sites, respectively. DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS [0136] The aspects described herein are not limited to specific embodiments, systems, compositions, methods, or configurations, and as such can, of course, vary. The terminology used herein is for the purpose of describing particular aspects only and, unless specifically defined herein, is not intended to be limiting. Compounds [0137] In one aspect, the present disclosure provides a compound of Formula (I): or a pharmaceutically acceptable salt thereof, wherein: Zn is an nth generation polyamidoamine (PAMAM) dendrimer; n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; each instance of Y is independently –ORA, –N(RA)2, or –C(=O)ORA; each instance of RA is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; each instance of L is independently optionally substituted alkylene, optionally substituted heterocyclylene, optionally substituted arylene, optionally substituted heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted alkylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted
U1197.70243WO00 34/193 #13587456v2 heteroarylene are independently replaced with –O–, –NR1–, –S–, –S(=O)–, –S(=O)2–, –C(=O)–, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene; each instance of R1 is independently hydrogen or optionally substituted alkyl; each instance of X is independently a conjugated agent; p is an integer between 1 and 2(n+2), inclusive; q is an integer between 0 and 2(n+2)-1, inclusive; and the sum of p and q is 2(n+2). [0138] In another aspect, the present disclosure provides a compound of Formula (I′): or a pharmaceutically acceptable salt thereof, wherein: Zn is an nth generation polyamidoamine (PAMAM) dendrimer; n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; each instance of Y is independently –ORA, –N(RA)2, or –C(=O)ORA; each instance of RA is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; each instance of L is independently optionally substituted alkylene, optionally substituted heterocyclylene, optionally substituted arylene, optionally substituted heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted alkylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene are independently replaced with –O–, –NR1–, –S–, –S(=O)–, –S(=O)2–, –C(=O)–, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene; each instance of R1 is independently hydrogen or optionally substituted alkyl; each instance of X is independently selected from formulae (ii) to (xi):
U1197.70243WO00 35/193 #13587456v2 p is an integer between 1 and 2(n+2), inclusive; q is an integer between 0 and 2(n+2)-1, inclusive; and the sum of p and q is 2(n+2). [0139] In another aspect, the present disclosure provides a compound of Formula (I′): or a pharmaceutically acceptable salt thereof, wherein: Zn is an nth generation polyamidoamine (PAMAM) dendrimer; n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; each instance of Y is independently –ORA, –N(RA)2, or –C(=O)ORA; each instance of RA is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl;
U1197.70243WO00 36/193 #13587456v2 each instance of L is independently optionally substituted alkylene, optionally substituted heterocyclylene, optionally substituted arylene, optionally substituted heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted alkylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene are independently replaced with –O–, –NR1–, –S–, –S(=O)–, –S(=O)2–, –C(=O)–, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene; each instance of R1 is independently hydrogen or optionally substituted alkyl; each instance of X is independently selected from formulae (ii) to (x): p is an integer between 1 and 2(n+2), inclusive;
U1197.70243WO00 37/193 #13587456v2 q is an integer between 0 and 2(n+2)-1, inclusive; and the sum of p and q is 2(n+2). [0140] In another aspect, the present disclosure provides a compound of Formula (II): or a pharmaceutically acceptable salt thereof, wherein: Zn is an nth generation polyamidoamine (PAMAM) dendrimer; n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; each instance of Y is independently –ORA, –N(RA)2, or –C(=O)ORA; each instance of RA is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; each instance of L is independently optionally substituted alkylene, optionally substituted heterocyclylene, optionally substituted arylene, optionally substituted heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted alkylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene are independently replaced with –O–, –NR1–, –S–, –S(=O)–, –S(=O)2–, –C(=O)–, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene; each instance of R1 is independently hydrogen or optionally substituted alkyl; p is an integer between 1 and 2(n+2), inclusive; q is an integer between 0 and 2(n+2)-1, inclusive; and the sum of p and q is 2(n+2). [0141] In another aspect, the present disclosure provides a compound of Formula (III): or a pharmaceutically acceptable salt thereof, wherein: Zn is an nth generation polyamidoamine (PAMAM) dendrimer; n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10;
U1197.70243WO00 38/193 #13587456v2 each instance of Y is independently –ORA, –N(RA)2, or –C(=O)ORA; each instance of RA is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; each instance of L is independently optionally substituted alkylene, optionally substituted heterocyclylene, optionally substituted arylene, optionally substituted heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted alkylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene are independently replaced with –O–, –NR1–, –S–, –S(=O)–, –S(=O)2–, –C(=O)–, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene; each instance of R1 is independently hydrogen or optionally substituted alkyl; p is an integer between 1 and 2(n+2), inclusive; q is an integer between 0 and 2(n+2)-1, inclusive; and the sum of p and q is 2(n+2). [0142] In another aspect, the present disclosure provides a compound of Formula (IV): or a pharmaceutically acceptable salt thereof, wherein: Zn is an nth generation polyamidoamine (PAMAM) dendrimer; n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; each instance of Y is independently –ORA, –N(RA)2, or –C(=O)ORA; each instance of RA is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; each instance of L is independently optionally substituted alkylene, optionally substituted heterocyclylene, optionally substituted arylene, optionally substituted heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted alkylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene are independently replaced with –O–, –NR1–, –S–, –S(=O)–, –S(=O)2–, –C(=O)–,
U1197.70243WO00 39/193 #13587456v2 optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene; each instance of R1 is independently hydrogen or optionally substituted alkyl; p is an integer between 1 and 2(n+2), inclusive; q is an integer between 0 and 2(n+2)-1, inclusive; and the sum of p and q is 2(n+2). [0143] In another aspect, the present disclosure provides a compound of Formula (V): or a pharmaceutically acceptable salt thereof, wherein: Zn is an nth generation polyamidoamine (PAMAM) dendrimer; n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; each instance of Y is independently –ORA, –N(RA)2, or –C(=O)ORA; each instance of RA is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; each instance of L is independently optionally substituted alkylene, optionally substituted heterocyclylene, optionally substituted arylene, optionally substituted heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted alkylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene are independently replaced with –O–, –NR1–, –S–, –S(=O)–, –S(=O)2–, –C(=O)–, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene; each instance of R1 is independently hydrogen or optionally substituted alkyl; p is an integer between 1 and 2(n+2), inclusive; q is an integer between 0 and 2(n+2)-1, inclusive; and the sum of p and q is 2(n+2). [0144] In another aspect, the present disclosure provides a compound of Formula (VI): or a pharmaceutically acceptable salt thereof, wherein: Zn is an nth generation polyamidoamine (PAMAM) dendrimer; n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10;
U1197.70243WO00 40/193 #13587456v2 each instance of Y is independently –ORA, –N(RA)2, or –C(=O)ORA; each instance of RA is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; each instance of L is independently optionally substituted alkylene, optionally substituted heterocyclylene, optionally substituted arylene, optionally substituted heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted alkylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene are independently replaced with –O–, –NR1–, –S–, –S(=O)–, –S(=O)2–, –C(=O)–, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene; each instance of R1 is independently hydrogen or optionally substituted alkyl; p is an integer between 1 and 2(n+2), inclusive; q is an integer between 0 and 2(n+2)-1, inclusive; and the sum of p and q is 2(n+2). [0145] In another aspect, the present disclosure provides a compound of Formula (VII): or a pharmaceutically acceptable salt thereof, wherein: Zn is an nth generation polyamidoamine (PAMAM) dendrimer; n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; each instance of Y is independently –ORA, –N(RA)2, or –C(=O)ORA; each instance of RA is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; each instance of L is independently optionally substituted alkylene, optionally substituted heterocyclylene, optionally substituted arylene, optionally substituted heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted alkylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene are independently replaced with –O–, –NR1–, –S–, –S(=O)–, –S(=O)2–, –C(=O)–,
U1197.70243WO00 41/193 #13587456v2 optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene; each instance of R1 is independently hydrogen or optionally substituted alkyl; p is an integer between 1 and 2(n+2), inclusive; q is an integer between 0 and 2(n+2)-1, inclusive; and the sum of p and q is 2(n+2). [0146] In another aspect, the present disclosure provides a compound of Formula (VIII): or a pharmaceutically acceptable salt thereof, wherein: Zn is an nth generation polyamidoamine (PAMAM) dendrimer; n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; each instance of Y is independently –ORA, –N(RA)2, or –C(=O)ORA; each instance of RA is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; each instance of L is independently optionally substituted alkylene, optionally substituted heterocyclylene, optionally substituted arylene, optionally substituted heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted alkylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene are independently replaced with –O–, –NR1–, –S–, –S(=O)–, –S(=O)2–, –C(=O)–, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene; each instance of R1 is independently hydrogen or optionally substituted alkyl; p is an integer between 1 and 2(n+2), inclusive; q is an integer between 0 and 2(n+2)-1, inclusive; and the sum of p and q is 2(n+2).
U1197.70243WO00 42/193 #13587456v2 [0147] In another aspect, the present disclosure provides a compound of Formula (IX): or a pharmaceutically acceptable salt thereof, wherein: Zn is an nth generation polyamidoamine (PAMAM) dendrimer; n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; each instance of Y is independently –ORA, –N(RA)2, or –C(=O)ORA; each instance of RA is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; each instance of L is independently optionally substituted alkylene, optionally substituted heterocyclylene, optionally substituted arylene, optionally substituted heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted alkylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene are independently replaced with –O–, –NR1–, –S–, –S(=O)–, –S(=O)2–, –C(=O)–, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene; each instance of R1 is independently hydrogen or optionally substituted alkyl; p is an integer between 1 and 2(n+2), inclusive; q is an integer between 0 and 2(n+2)-1, inclusive; and the sum of p and q is 2(n+2).
U1197.70243WO00 43/193 #13587456v2 [0148] In another aspect, the present disclosure provides a compound of Formula (X): or a pharmaceutically acceptable salt thereof, wherein: Zn is an nth generation polyamidoamine (PAMAM) dendrimer; n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; each instance of Y is independently –ORA, –N(RA)2, or –C(=O)ORA; each instance of RA is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; each instance of L is independently optionally substituted alkylene, optionally substituted heterocyclylene, optionally substituted arylene, optionally substituted heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted alkylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene are independently replaced with –O–, –NR1–, –S–, –S(=O)–, –S(=O)2–, –C(=O)–, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene; each instance of R1 is independently hydrogen or optionally substituted alkyl; p is an integer between 1 and 2(n+2), inclusive; q is an integer between 0 and 2(n+2)-1, inclusive; and the sum of p and q is 2(n+2). [0149] In another aspect, the present disclosure provides a compound of Formula (XI): or a pharmaceutically acceptable salt thereof, wherein: Zn is an nth generation polyamidoamine (PAMAM) dendrimer;
U1197.70243WO00 44/193 #13587456v2 n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; each instance of Y is independently –ORA, –N(RA)2, or –C(=O)ORA; each instance of RA is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; each instance of L is independently optionally substituted alkylene, optionally substituted heterocyclylene, optionally substituted arylene, optionally substituted heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted alkylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene are independently replaced with –O–, –NR1–, –S–, –S(=O)–, –S(=O)2–, –C(=O)–, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene; each instance of R1 is independently hydrogen or optionally substituted alkyl; p is an integer between 1 and 2(n+2), inclusive; q is an integer between 0 and 2(n+2)-1, inclusive; and the sum of p and q is 2(n+2). Zn, n, p, and q [0150] As generally described herein, Zn is an nth generation polyamidoamine (PAMAM) dendrimer. [0151] In some embodiments, the nth generation PAMAM dendrimer is prepared by a two-step iterative reaction sequence that produces concentric shells (generations) of dendritic 0-alanine units around a central initiator core. In some embodiments, the nth generation PAMAM dendrimer is a generation 1 PAMAM dendrimer, generation 2 PAMAM dendrimer, generation 3 PAMAM dendrimer, generation 4 PAMAM dendrimer, generation 5 PAMAM dendrimer, generation 6 PAMAM dendrimer, generation 7 PAMAM dendrimer, generation 8 PAMAM dendrimer, generation 9 PAMAM dendrimer, or generation 10 PAMAM dendrimer. In some embodiments, the nth generation PAMAM dendrimer is a generation 6 PAMAM dendrimer. [0152] In some embodiments, Zn is an nth generation PAMAM dendrimer (i.e., Zn is the core of an nth generation PAMAM dendrimer, and does not include the terminal groups of the dendrimer). By means of non-limiting example, in some embodiments, Z0 is a 0th generation dendrimer (i.e., Z0 is the core of a 0th generation PAMAM dendrimer). In some embodiments, Z0 is a 0th generation dendrimer (i.e., Z0 is the core of a 0th generation PAMAM dendrimer) of formula:
U1197.70243WO00 45/193 #13587456v2 . [0153] By means of an additional non-limiting example, in some embodiments, Z1 is a 1st generation PAMAM dendrimer (i.e., Z1 is the core of a 1st generation PAMAM dendrimer). In some embodiments, Z1 is a 1st generation PAMAM dendrimer (i.e., Z1 is the core of a 1st generation PAMAM dendrimer) of formula: . [0154] As a further non-limiting example, the compound of Formula (I) or Formula (I′), wherein: Z1 is an 1st generation polyamidoamine (PAMAM) dendrimer; n is 1; each instance of Y is independently can be represented by the formula: ,
U1197.70243WO00 46/193 #13587456v2 or a pharmaceutically acceptable salt thereof. [0155] As generally described herein, n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. [0156] In some embodiments, n is 0, 1, 2, 3, 4, 5, 6, 7, 8, or 9. In some embodiments, n is 0, 1, 2, 3, 4, 5, 6, 7, or 8. In some embodiments, n is 0, 1, 2, 3, 4, 5, 6, or 7. In some embodiments, n is 0, 1, 2, 3, 4, 5, or 6. In some embodiments, n is 0, 1, 2, 3, 4, or 5. In some embodiments, n is 0, 1, 2, 3, or 4. In some embodiments, n is 0, 1, 2, or 3. In some embodiments, n is 0, 1, or 2. In some embodiments, n is 0 or 1. In some embodiments, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, n is 1, 2, 3, 4, 5, 6, 7, 8, or 9. In some embodiments, n is 1, 2, 3, 4, 5, 6, 7, or 8. In some embodiments, n is 1, 2, 3, 4, 5, 6, or 7. In some embodiments, n is 1, 2, 3, 4, 5, or 6. In some embodiments, n is 1, 2, 3, 4, or 5. In some embodiments, n is 1, 2, 3, or 4. In some embodiments, n is 1, 2 or 3. In some embodiments, n is 1 or 2. In some embodiments, n is 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, n is 2, 3, 4, 5, 6, 7, 8, or 9. In some embodiments, n is 2, 3, 4, 5, 6, 7, or 8. In some embodiments, n is 2, 3, 4, 5, 6, or 7. In some embodiments, n is 2, 3, 4, 5, or 6. In some embodiments, n is 2, 3, 4, or 5. In some embodiments, n is 2, 3, or 4. In some embodiments, n is 2 or 3. In some embodiments, n is 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, n is 3, 4, 5, 6, 7, 8, or 9. In some embodiments, n is 3, 4, 5, 6, 7, or 8. In some embodiments, n is 3, 4, 5, 6, or 7. In some embodiments, n is 3, 4, 5, or 6. In some embodiments, n is 3, 4, or 5. In some embodiments, n is 3 or 4. [0157] In some embodiments, n is 4 or greater. In some embodiments, n is 4, 5, 6, 7, 8, 9, or 10. In some embodiments, n is 4, 5, 6, 7, 8, or 9. In some embodiments, n is 4, 5, 6, 7, or 8. In some embodiments, n is 4, 5, 6, or 7. In some embodiments, n is 4, 5, or 6. In some embodiments, n is 4 or 5. In some embodiments, n is 5 or greater. In some embodiments, n is 5, 6, 7, 8, 9, or 10. In some embodiments, n is 5, 6, 7, 8, or 9. In some embodiments, n is 5, 6, 7, or 8. In some embodiments, n is 5, 6, or 7. In some embodiments, n is 5 or 6. In some embodiments, n is 6 or greater. In some embodiments, n is 6, 7, 8, 9, or 10. In some embodiments, n is 6, 7, 8, or 9. In some embodiments, n is 6, 7, or 8. In some embodiments, n is 6 or 7. In some embodiments, n is 7 or greater. In some embodiments, n is 7, 8, 9, or 10. In some embodiments, n is 7, 8, or 9. In some embodiments, n is 7 or 8. In some embodiments, n is 8 or greater. In some embodiments, n is 8, 9, or 10. In some embodiments, n is 8 or 9. In some embodiments, n is 9 or greater. In some embodiments, n is 9 or 10. [0158] In some embodiments, n is 0. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5. In some embodiments, n is 6. In some embodiments, n is 7. In some embodiments, n is 8. In some embodiments, n is 9. In some embodiments, n is 10. [0159] In some embodiments, n is 0, such that Zn is Z0, and Z0 is a 0th generation PAMAM dendrimer. In some embodiments, n is 1, such that Zn is Z1, and Z1 is a 1st generation PAMAM dendrimer. In some embodiments, n is 2, such that Zn is Z2, and Z2 is a 2nd generation PAMAM dendrimer. In some embodiments, n is 3, such that Zn is Z3, and Z3 is a 3rd generation PAMAM dendrimer. In some embodiments, n is 4, such that Zn is Z4, and Z4 is a 4th generation PAMAM
U1197.70243WO00 47/193 #13587456v2 dendrimer. In some embodiments, n is 5, such that Zn is Z5, and Z5 is a 5th generation PAMAM dendrimer. In some embodiments, n is 6, such that Zn is Z6, and Z6 is a 6th generation PAMAM dendrimer. In some embodiments, n is 7, such that Zn is Z7, and Z7 is a 7th generation PAMAM dendrimer. In some embodiments, n is 8, such that Zn is Z8, and Z8 is a 8th generation PAMAM dendrimer. In some embodiments, n is 9, such that Zn is Z9, and Z9 is a 9th generation PAMAM dendrimer. In some embodiments, n is 10, such that Zn is Z10, and Z10 is a 10th generation PAMAM dendrimer. [0160] In some embodiments, Zn is Z4, Z5, Z6, Z7, Z8, Z9, or Z10. In some embodiments, Zn is Z4, Z5, Z6, Z7, Z8, or Z9. In some embodiments, Zn is Z4, Z5, Z6, Z7, or Z8. In some embodiments, Zn is Z4, Z5, Z6, or Z7. In some embodiments, Zn is Z4, Z5, or Z6. In some embodiments, Zn is Z5, Z6, Z7, Z8, Z9, or Z10. In some embodiments, Zn is Z5, Z6, Z7, Z8, or Z9. In some embodiments, Zn is Z5, Z6, Z7, or Z8. In some embodiments, Zn is Z5, Z6, or Z7. In some embodiments, Zn is Z5 or Z6. In some embodiments, Zn is Z6, Z7, Z8, Z9, or Z10. In some embodiments, Zn is Z6, Z7, Z8, or Z9. In some embodiments, Zn is Z6, Z7, or Z8. In some embodiments, Zn is Z6 or Z7. [0161] As generally described herein, p is an integer between 1 and 2(n+2), inclusive; q is an integer between 0 and 2(n+2)-1, inclusive; and the sum of p and q is 2(n+2). [0162] In some embodiments, n is 0, such that the sum of p and q is 4. In some embodiments, n is 1, such that the sum of p and q is 8. In some embodiments, n is 2, such that the sum of p and q is 16. In some embodiments, n is 3, such that the sum of p and q is 32. [0163] In some embodiments, n is 4, such that the sum of p and q is 64. In some embodiments, the sum of p and q is 64. In some embodiments, the sum of p and q is 64, p is an integer between 1 and 10, inclusive, and q is an integer between 54 and 63, inclusive. In some embodiments, the sum of p and q is 64, p is an integer between 1 and 5, inclusive, and q is an integer between 59 and 63, inclusive. [0164] In some embodiments, n is 5, such that the sum of p and q is 128. In some embodiments, the sum of p and q is 128. In some embodiments, the sum of p and q is 128, p is an integer between 1 and 20, inclusive, and q is an integer between 108 and 127, inclusive. In some embodiments, the sum of p and q is 128, p is an integer between 1 and 15, inclusive, and q is an integer between 113 and 127, inclusive. In some embodiments, the sum of p and q is 128, p is an integer between 1 and 10, inclusive, and q is an integer between 118 and 127, inclusive. In some embodiments, the sum of p and q is 128, p is an integer between 1 and 5, inclusive, and q is an integer between 123 and 127, inclusive. [0165] In some embodiments, n is 6, such that the sum of p and q is 256. In some embodiments, the sum of p and q is 256. In some embodiments, the sum of p and q is 256, p is an integer between 1 and 40, inclusive, and q is an integer between 216 and 255, inclusive. In some embodiments, the sum of p and q is 256, p is an integer between 5 and 35, inclusive, and q is an integer between 221 and 251, inclusive. In some embodiments, the sum of p and q is 256, p is an integer between 8 and 32,
U1197.70243WO00 48/193 #13587456v2 inclusive, and q is an integer between 224 and 248, inclusive. In some embodiments, the sum of p and q is 256, p is an integer between 10 and 30, inclusive, and q is an integer between 226 and 246, inclusive. In some embodiments, the sum of p and q is 256, p is an integer between 12 and 28, inclusive, and q is an integer between 228 and 244, inclusive. In some embodiments, the sum of p and q is 256, p is an integer between 14 and 26, inclusive, and q is an integer between 230 and 242, inclusive. In some embodiments, the sum of p and q is 256, p is an integer between 15 and 25, inclusive, and q is an integer between 231 and 241, inclusive. In some embodiments, the sum of p and q is 256, p is an integer between 16 and 24, inclusive, and q is an integer between 232 and 240, inclusive. In some embodiments, the sum of p and q is 256, p is an integer between 17 and 23, inclusive, and q is an integer between 233 and 239, inclusive. In some embodiments, the sum of p and q is 256, p is an integer between 18 and 22, inclusive, and q is an integer between 234 and 238, inclusive. In some embodiments, the sum of p and q is 256, p is an integer between 19 and 21, inclusive, and q is an integer between 235 and 237, inclusive. [0166] In some embodiments, the sum of p and q is 256, p is an integer between 1 and 30, inclusive, and q is an integer between 226 and 255, inclusive. In some embodiments, the sum of p and q is 256, p is an integer between 3 and 27, inclusive, and q is an integer between 229 and 253, inclusive. In some embodiments, the sum of p and q is 256, p is an integer between 5 and 25, inclusive, and q is an integer between 231 and 251, inclusive. In some embodiments, the sum of p and q is 256, p is an integer between 7 and 23, inclusive, and q is an integer between 233 and 249, inclusive. In some embodiments, the sum of p and q is 256, p is an integer between 9 and 21, inclusive, and q is an integer between 235 and 247, inclusive. In some embodiments, the sum of p and q is 256, p is an integer between 10 and 20, inclusive, and q is an integer between 236 and 246, inclusive. In some embodiments, the sum of p and q is 256, p is an integer between 11 and 19, inclusive, and q is an integer between 237 and 245, inclusive. In some embodiments, the sum of p and q is 256, p is an integer between 12 and 18, inclusive, and q is an integer between 238 and 244, inclusive. In some embodiments, the sum of p and q is 256, p is an integer between 13 and 17, inclusive, and q is an integer between 239 and 243, inclusive. In some embodiments, the sum of p and q is 256, p is an integer between 14 and 16, inclusive, and q is an integer between 240 and 242, inclusive. [0167] In some embodiments, p is 1 and q is 255. In some embodiments, p is 2 and q is 254. In some embodiments, p is 3 and q is 253. In some embodiments, p is 4 and q is 252. In some embodiments, p is 5 and q is 251. In some embodiments, p is 6 and q is 250. In some embodiments, p is 7 and q is 249. In some embodiments, p is 8 and q is 248. In some embodiments, p is 9 and q is 247. In some embodiments, p is 10 and q is 246. In some embodiments, p is 11 and q is 245. In some embodiments, p is 12 and q is 244. In some embodiments, p is 13 and q is 243. In some embodiments, p is 14 and q is 242. In some embodiments, p is 15 and q is 241. In some embodiments, p is 16 and q is 240. In some embodiments, p is 17 and q is 239. In some embodiments, p is 18 and q is 238. In some embodiments, p is 19 and q is 237. In some embodiments, p is 20 and q is 236. In some embodiments,
U1197.70243WO00 49/193 #13587456v2 p is 21 and q is 235. In some embodiments, p is 22 and q is 234. In some embodiments, p is 23 and q is 233. In some embodiments, p is 24 and q is 232. In some embodiments, p is 25 and q is 231. In some embodiments, p is 26 and q is 230. In some embodiments, p is 27 and q is 229. In some embodiments, p is 28 and q is 228. In some embodiments, p is 29 and q is 227. In some embodiments, p is 30 and q is 226. In some embodiments, p is 31 and q is 225. In some embodiments, p is 32 and q is 224. In some embodiments, p is 33 and q is 223. In some embodiments, p is 34 and q is 222. In some embodiments, p is 35 and q is 221. In some embodiments, p is 36 and q is 220. In some embodiments, p is 37 and q is 219. In some embodiments, p is 38 and q is 218. In some embodiments, p is 39 and q is 217. In some embodiments, p is 40 and q is 216. In some embodiments, p is 15 and q is 241; or p is 20 and q is 236. [0168] In some embodiments, n is 7, such that the sum of p and q is 512. In some embodiments, the sum of p and q is 512. In some embodiments, the sum of p and q is 512, p is an integer between 1 and 80, inclusive, and q is an integer between 432 and 511, inclusive. In some embodiments, the sum of p and q is 512, p is an integer between 1 and 40, inclusive, and q is an integer between 472 and 511, inclusive. [0169] In some embodiments, n is 8, such that the sum of p and q is 1024. In some embodiments, the sum of p and q is 1024. In some embodiments, the sum of p and q is 1024, p is an integer between 1 and 160, inclusive, and q is an integer between 864 and 1023, inclusive. In some embodiments, the sum of p and q is 512, p is an integer between 1 and 80, inclusive, and q is an integer between 944 and 1023, inclusive. [0170] In some embodiments, n is 9, such that the sum of p and q is 2048. In some embodiments, the sum of p and q is 2048. In some embodiments, the sum of p and q is 2048, p is an integer between 1 and 320, inclusive, and q is an integer between 1728 and 2047, inclusive. In some embodiments, the sum of p and q is 2048, p is an integer between 1 and 160, inclusive, and q is an integer between 1888 and 2047, inclusive. [0171] In some embodiments, n is 10, such that the sum of p and q is 4096. In some embodiments, the sum of p and q is 4096. In some embodiments, the sum of p and q is 4096, p is an integer between 1 and 640, inclusive, and q is an integer between 3456 and 4095, inclusive. In some embodiments, the sum of p and q is 4096, p is an integer between 1 and 320, inclusive, and q is an integer between 3776 and 4095, inclusive. [0172] In some embodiments, p is at least 1. In some embodiments, p is at least 2. In some embodiments, p is at least 3. In some embodiments, p is at least 4. In some embodiments, p is at least 5. In some embodiments, p is at least 6. In some embodiments, p is at least 7. In some embodiments, p is at least 8. In some embodiments, p is at least 9. In some embodiments, p is at least 10. In some embodiments, p is at least 11. In some embodiments, p is at least 12. In some embodiments, p is at least 13. In some embodiments, p is at least 14. In some embodiments, p is at least 15. In some
U1197.70243WO00 50/193 #13587456v2 embodiments, p is at least 16. In some embodiments, p is at least 17. In some embodiments, p is at least 18. In some embodiments, p is at least 19. In some embodiments, p is at least 20. L and R1 [0173] As generally described herein, each instance of L is independently optionally substituted alkylene, optionally substituted heterocyclylene, optionally substituted arylene, optionally substituted heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted alkylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene are independently replaced with –O–, –NR1–, –S–, – S(=O)–, –S(=O)2–, –C(=O)–, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene. [0174] In some embodiments, at least one instance of L is optionally substituted alkylene, optionally substituted heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted alkylene or optionally substituted heteroarylene are independently replaced with –O–, –NR1–, –S–, –S(=O)–, –S(=O)2–, –C(=O)–, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene. In some embodiments, at least one instance of L is optionally substituted alkylene, optionally substituted heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted alkylene or optionally substituted heteroarylene are independently replaced with –O–, –NR1–, –C(=O)–, or optionally substituted heteroarylene. In some embodiments, at least one instance of L is optionally substituted alkylene, optionally substituted heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted alkylene are independently replaced with –O–, –NR1–, –C(=O)–, or optionally substituted heteroarylene. [0175] In some embodiments, at least one instance of L is optionally substituted C1-20 alkylene, optionally substituted 5-6 membered heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted C1-20 alkylene or optionally substituted 5- 6 membered heteroarylene are independently replaced with –O–, –NR1–, –S–, –S(=O)–, –S(=O)2–, – C(=O)–, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene. In some embodiments, at least one instance of L is optionally substituted C1-20 alkylene, optionally substituted 5-6 membered heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted C1-20 alkylene or optionally substituted 5-6 membered heteroarylene are independently replaced with –O–, –NR1–, –C(=O)–, or optionally substituted heteroarylene. In some embodiments, at least one instance of L is optionally substituted C1-20 alkylene, optionally substituted 5-6 membered heteroarylene, or a combination thereof, optionally wherein one or more backbone
U1197.70243WO00 51/193 #13587456v2 carbon atoms in the optionally substituted C1-20 alkylene are independently replaced with –O–, –NR1–, –C(=O)–, or optionally substituted heteroarylene. [0176] In some embodiments, at least one instance of L is optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted linear C1- 20 alkylene or optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms are independently replaced with –O–, –NR1–, –S–, –S(=O)–, –S(=O)2–, –C(=O)–, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene. In some embodiments, at least one instance of L is optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted linear C1-20 alkylene or optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms are independently replaced with –O–, –NR1–, –C(=O)–, or optionally substituted heteroarylene. In some embodiments, at least one instance of L is optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted linear C1-20 alkylene are independently replaced with –O–, –NR1–, –C(=O)–, or optionally substituted heteroarylene. [0177] In some embodiments, at least one instance of L is optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein one or more backbone carbon atoms in the optionally substituted linear C1-20 alkylene are independently replaced with –O–, –NR1–, –C(=O)–, or optionally substituted heteroarylene. In some embodiments, at least one instance of L is optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1, 2, 3, 4, or 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –O–. In some embodiments, at least one instance of L is optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1 backbone carbon atom in the optionally substituted linear C1-20 alkylene is replaced with –O–. In some embodiments, at least one instance of L is optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 2 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –O–. In some embodiments, at least one instance of L is optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 3 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –O–. In some embodiments, at least one instance of L is optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof,
U1197.70243WO00 52/193 #13587456v2 wherein at least 4 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –O–. In some embodiments, at least one instance of L is optionally substituted linear C1- 20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 5 backbone carbon atoms in the optionally substituted linear C1- 20 alkylene are replaced with –O–. [0178] In some embodiments, at least one instance of L is optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1, 2, 3, 4, or 5 backbone carbon atoms in the optionally substituted linear C1- 20 alkylene are replaced with –NR1–. In some embodiments, at least one instance of L is optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1 backbone carbon atoms in the optionally substituted linear C1-20 alkylene is replaced with –NR1–. In some embodiments, at least one instance of L is optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 2 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –NR1–. In some embodiments, at least one instance of L is optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 3 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –NR1–. In some embodiments, at least one instance of L is optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 4 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –NR1–. In some embodiments, at least one instance of L is optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 5 backbone carbon atoms in the optionally substituted linear C1- 20 alkylene are replaced with –NR1–. [0179] In some embodiments, at least one instance of L is optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1, 2, 3, 4, or 5 backbone carbon atoms in the optionally substituted linear C1- 20 alkylene are replaced with –C(=O)–. In some embodiments, at least one instance of L is optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1 backbone carbon atom in the optionally substituted linear C1-20 alkylene is replaced with –C(=O)–. In some embodiments, at least one instance of L is optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 2 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –C(=O)–. In some embodiments, at least one instance of L is optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 3
U1197.70243WO00 53/193 #13587456v2 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –C(=O)–. In some embodiments, at least one instance of L is optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 4 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –C(=O)–. In some embodiments, at least one instance of L is optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –C(=O)–. [0180] In some embodiments, at least one instance of L is optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1, 2, 3, 4, or 5 backbone carbon atoms in the optionally substituted linear C1- 20 alkylene are replaced with optionally substituted heteroarylene. In some embodiments, at least one instance of L is optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1 backbone carbon atom in the optionally substituted linear C1-20 alkylene is replaced with optionally substituted heteroarylene. [0181] In some embodiments, at least one instance of L comprises optionally substituted alkylene, optionally substituted heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted alkylene or optionally substituted heteroarylene are independently replaced with –O–, –NR1–, –S–, –S(=O)–, –S(=O)2–, –C(=O)–, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene. In some embodiments, at least one instance of L comprises optionally substituted alkylene, optionally substituted heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted alkylene or optionally substituted heteroarylene are independently replaced with –O–, –NR1–, –C(=O)–, or optionally substituted heteroarylene. In some embodiments, at least one instance of L comprises optionally substituted alkylene, optionally substituted heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted alkylene are independently replaced with –O–, –NR1–, –C(=O)–, or optionally substituted heteroarylene. [0182] In some embodiments, at least one instance of L comprises optionally substituted C1-20 alkylene, optionally substituted 5-6 membered heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted C1-20 alkylene or optionally substituted 5-6 membered heteroarylene are independently replaced with –O–, –NR1–, –S–, –S(=O)–, –S(=O)2–, –C(=O)–, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene. In some embodiments, at least one instance of L comprises optionally substituted C1-20 alkylene, optionally substituted 5-6 membered heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in
U1197.70243WO00 54/193 #13587456v2 the optionally substituted C1-20 alkylene or optionally substituted 5-6 membered heteroarylene are independently replaced with –O–, –NR1–, –C(=O)–, or optionally substituted heteroarylene. In some embodiments, at least one instance of L comprises optionally substituted C1-20 alkylene, optionally substituted 5-6 membered heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted C1-20 alkylene are independently replaced with – O–, –NR1–, –C(=O)–, or optionally substituted heteroarylene. [0183] In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted linear C1-20 alkylene or optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms are independently replaced with –O–, –NR1–, –S–, –S(=O)–, –S(=O)2–, –C(=O)–, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted linear C1-20 alkylene or optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms are independently replaced with –O–, –NR1–, –C(=O)–, or optionally substituted heteroarylene. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted linear C1-20 alkylene are independently replaced with –O–, –NR1–, –C(=O)–, or optionally substituted heteroarylene. [0184] In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein one or more backbone carbon atoms in the optionally substituted linear C1-20 alkylene are independently replaced with –O–, –NR1–, –C(=O)–, or optionally substituted heteroarylene. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1, 2, 3, 4, or 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –O–. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1 backbone carbon atom in the optionally substituted linear C1-20 alkylene is replaced with –O–. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 2 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –O–. In some embodiments, at least one instance of L comprises optionally
U1197.70243WO00 55/193 #13587456v2 substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 3 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –O–. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 4 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –O–. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –O–. [0185] In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1, 2, 3, 4, or 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –NR1–. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1 backbone carbon atoms in the optionally substituted linear C1-20 alkylene is replaced with –NR1–. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 2 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –NR1–. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 3 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –NR1–. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 4 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –NR1–. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –NR1–. [0186] In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1, 2, 3, 4, or 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –C(=O)–. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1
U1197.70243WO00 56/193 #13587456v2 backbone carbon atom in the optionally substituted linear C1-20 alkylene is replaced with –C(=O)–. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 2 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –C(=O)–. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 3 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –C(=O)–. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 4 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –C(=O)–. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –C(=O)–. [0187] In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1, 2, 3, 4, or 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with optionally substituted heteroarylene. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1 backbone carbon atom in the optionally substituted linear C1-20 alkylene is replaced with optionally substituted heteroarylene. [0188] In some embodiments, at least one instance of L comprises optionally substituted alkylene, optionally wherein one or more backbone carbon atoms in the optionally substituted alkylene are independently replaced with –O–, –NR1–, –S–, –S(=O)–, –S(=O)2–, –C(=O)–, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene. In some embodiments, at least one instance of L comprises optionally substituted alkylene, optionally wherein one or more backbone carbon atoms in the optionally substituted alkylene are independently replaced with –O–, –NR1–, –C(=O)–, or optionally substituted heteroarylene. [0189] In some embodiments, at least one instance of L comprises optionally substituted C1-20 alkylene, optionally wherein one or more backbone carbon atoms in the optionally substituted C1-20 alkylene are independently replaced with –O–, –NR1–, –S–, –S(=O)–, –S(=O)2–, –C(=O)–, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene. In some embodiments, at least one instance of L comprises optionally substituted C1-20 alkylene, optionally wherein one or more backbone carbon atoms in the
U1197.70243WO00 57/193 #13587456v2 optionally substituted C1-20 alkylene are independently replaced with –O–, –NR1–, –C(=O)–, or optionally substituted heteroarylene. [0190] In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, optionally wherein one or more backbone carbon atoms in the optionally substituted linear C1-20 alkylene are independently replaced with –O–, –NR1–, –S–, –S(=O)–, –S(=O)2–, –C(=O)–, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, optionally wherein one or more backbone carbon atoms in the optionally substituted linear C1-20 alkylene are independently replaced with –O–, – NR1–, –C(=O)–, or optionally substituted heteroarylene. [0191] In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, wherein one or more backbone carbon atoms in the optionally substituted linear C1-20 alkylene are independently replaced with –O–, –NR1–, –C(=O)–, or optionally substituted heteroarylene. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, wherein at least 1, 2, 3, 4, or 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –O–. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, wherein at least 1 backbone carbon atom in the optionally substituted linear C1-20 alkylene is replaced with –O–. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, wherein at least 2 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –O–. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, wherein at least 3 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with – O–. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, wherein at least 4 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –O–. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, wherein at least 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –O–. [0192] In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, wherein at least 1, 2, 3, 4, or 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –NR1–. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, wherein at least 1 backbone carbon atoms in the optionally substituted linear C1-20 alkylene is replaced with –NR1–. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, wherein at least 2 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –NR1–. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, wherein at least 3 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –NR1–. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene,
U1197.70243WO00 58/193 #13587456v2 wherein at least 4 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –NR1–. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, wherein at least 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –NR1–. [0193] In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, wherein at least 1, 2, 3, 4, or 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –C(=O)–. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, wherein at least 1 backbone carbon atom in the optionally substituted linear C1-20 alkylene is replaced with –C(=O)–. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, wherein at least 2 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –C(=O)–. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, wherein at least 3 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –C(=O)–. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, wherein at least 4 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –C(=O)–. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, wherein at least 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –C(=O)–. [0194] In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, wherein at least 1, 2, 3, 4, or 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with optionally substituted heteroarylene. In some embodiments, at least one instance of L comprises optionally substituted linear C1-20 alkylene, wherein at least 1 backbone carbon atom in the optionally substituted linear C1-20 alkylene is replaced with optionally substituted heteroarylene. [0195] In some embodiments, at least one instance of L comprises ,
U1197.70243WO00 59/193 #13587456v2 , wherein each x is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; and each y is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, at least one instance of L comprises 10; and each y is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, at least one instance of L . In some embodiments, at least one instance of L
U1197.70243WO00 60/193 #13587456v2 [0196] In some embodiments, at least one instance of L comprises optionally substituted heteroarylene, optionally wherein one or more backbone carbon atoms in the optionally substituted heteroarylene are independently replaced with –O–, –NR1–, –S–, –S(=O)–, –S(=O)2–, –C(=O)–, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene. In some embodiments, at least one instance of L comprises optionally substituted heteroarylene, optionally wherein one or more backbone carbon atoms in the optionally substituted heteroarylene are independently replaced with –O–, –NR1–, – C(=O)–, or optionally substituted heteroarylene. [0197] In some embodiments, at least one instance of L comprises optionally substituted heteroarylene. In some embodiments, at least one instance of L comprises optionally substituted 5-6 membered heteroarylene. In some embodiments, at least one instance of L comprises optionally substituted 5 membered heteroarylene. In some embodiments, at least one instance of L comprises optionally substituted 6 membered heteroarylene. In some embodiments, at least one instance of L comprises optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring heteroatoms selected from O, N, and S. In some embodiments, at least one instance of L comprises optionally substituted 5 membered heteroarylene having 1, 2, or 3 ring heteroatoms selected from O, N, and S. In some embodiments, at least one instance of L comprises optionally substituted 6 membered heteroarylene having 1, 2, or 3 ring heteroatoms selected from O, N, and S. [0198] In some embodiments, at least one instance of L comprises optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms. In some embodiments, at least one instance of L comprises optionally substituted 5-6 membered heteroarylene having 1 ring N atom. In some embodiments, at least one instance of L comprises optionally substituted 5-6 membered heteroarylene having 2 ring N atoms. In some embodiments, at least one instance of L comprises optionally substituted 5-6 membered heteroarylene having 3 ring N atoms. In some embodiments, at least one instance of L comprises optionally substituted 5 membered heteroarylene having 1, 2, or 3 ring N atoms. In some embodiments, at least one instance of L comprises optionally substituted 5 membered heteroarylene having 1 ring N atom. In some embodiments, at least one instance of L comprises optionally substituted 5 membered heteroarylene having 2 ring N atoms. In some embodiments, at least one instance of L comprises optionally substituted 5 membered heteroarylene having 3 ring N atoms. In some embodiments, at least one instance of L comprises optionally substituted 6 membered heteroarylene having 1, 2, or 3 ring N atoms. In some embodiments, at least one instance of L comprises optionally substituted 6 membered heteroarylene having 1 ring N atom. In some embodiments, at least one instance of L comprises optionally substituted 6 membered heteroarylene having 2 ring N atoms. In some embodiments, at least one instance of L comprises optionally substituted 6 membered heteroarylene having 3 ring N atoms.
U1197.70243WO00 61/193 #13587456v2 [0199] In some embodiments, at least one instance of L comprises , , . In some embodiments, at least one instance of L comprises . In some embodiments, at least one instance of L comprises . In some embodiments, at least one instance of L comprises . In some embodiments, at least one instance of L is , [0201] In some embodiments, at least one instance of L comprises
U1197.70243WO00 62/193 #13587456v2 . [0202] In some embodiments, each instance of L independently comprises optionally substituted alkylene, optionally substituted heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted alkylene or optionally substituted heteroarylene are independently replaced with –O–, –NR1–, –S–, –S(=O)–, –S(=O)2–, –C(=O)–, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene. In some embodiments, each instance of L independently comprises optionally substituted alkylene, optionally substituted heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted alkylene or optionally substituted heteroarylene are independently replaced with –O–, – NR1–, –C(=O)–, or optionally substituted heteroarylene. In some embodiments, each instance of L independently comprises optionally substituted alkylene, optionally substituted heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted alkylene are independently replaced with –O–, –NR1–, –C(=O)–, or optionally substituted heteroarylene. [0203] In some embodiments, each instance of L independently comprises optionally substituted C1- 20 alkylene, optionally substituted 5-6 membered heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted C1-20 alkylene or optionally substituted 5-6 membered heteroarylene are independently replaced with –O–, –NR1–, –S–, –S(=O)–, –S(=O)2–, –C(=O)–, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene. In some embodiments, each instance of L independently comprises optionally substituted C1-20 alkylene, optionally substituted 5-6 membered heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted C1-20 alkylene or optionally substituted 5-6 membered heteroarylene are independently replaced with –O–, –NR1–, –C(=O)–, or optionally substituted heteroarylene. In some embodiments, each instance of L independently comprises optionally substituted C1-20 alkylene, optionally substituted 5-6 membered heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted C1-20 alkylene are independently replaced with –O–, –NR1–, –C(=O)–, or optionally substituted heteroarylene. [0204] In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted linear C1-20 alkylene or optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms are independently replaced with –O–, –NR1–, –S–, –S(=O)–, –S(=O)2–,
U1197.70243WO00 63/193 #13587456v2 –C(=O)–, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted linear C1-20 alkylene or optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms are independently replaced with –O–, –NR1–, –C(=O)–, or optionally substituted heteroarylene. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted linear C1-20 alkylene are independently replaced with –O–, –NR1–, –C(=O)–, or optionally substituted heteroarylene. [0205] In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein one or more backbone carbon atoms in the optionally substituted linear C1-20 alkylene are independently replaced with –O–, –NR1–, –C(=O)–, or optionally substituted heteroarylene. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1, 2, 3, 4, or 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –O–. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1 backbone carbon atom in the optionally substituted linear C1-20 alkylene is replaced with –O–. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 2 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –O–. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 3 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –O–. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 4 backbone carbon atoms in the optionally substituted linear C1- 20 alkylene are replaced with –O–. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –O–.
U1197.70243WO00 64/193 #13587456v2 [0206] In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1, 2, 3, 4, or 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –NR1–. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1 backbone carbon atoms in the optionally substituted linear C1-20 alkylene is replaced with –NR1–. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 2 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –NR1–. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 3 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –NR1–. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 4 backbone carbon atoms in the optionally substituted linear C1- 20 alkylene are replaced with –NR1–. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –NR1–. [0207] In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1, 2, 3, 4, or 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –C(=O)–. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1 backbone carbon atom in the optionally substituted linear C1-20 alkylene is replaced with –C(=O)–. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 2 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –C(=O)–. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 3 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –C(=O)–. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a
U1197.70243WO00 65/193 #13587456v2 combination thereof, wherein at least 4 backbone carbon atoms in the optionally substituted linear C1- 20 alkylene are replaced with –C(=O)–. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –C(=O)–. [0208] In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1, 2, 3, 4, or 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with optionally substituted heteroarylene. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms, or a combination thereof, wherein at least 1 backbone carbon atom in the optionally substituted linear C1-20 alkylene is replaced with optionally substituted heteroarylene. [0209] In some embodiments, each instance of L independently comprises optionally substituted alkylene, optionally wherein one or more backbone carbon atoms in the optionally substituted alkylene are independently replaced with –O–, –NR1–, –S–, –S(=O)–, –S(=O)2–, –C(=O)–, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene. In some embodiments, each instance of L independently comprises optionally substituted alkylene, optionally wherein one or more backbone carbon atoms in the optionally substituted alkylene are independently replaced with –O–, –NR1–, –C(=O)–, or optionally substituted heteroarylene. [0210] In some embodiments, each instance of L independently comprises optionally substituted C1- 20 alkylene, optionally wherein one or more backbone carbon atoms in the optionally substituted C1-20 alkylene are independently replaced with –O–, –NR1–, –S–, –S(=O)–, –S(=O)2–, –C(=O)–, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene. In some embodiments, each instance of L independently comprises optionally substituted C1-20 alkylene, optionally wherein one or more backbone carbon atoms in the optionally substituted C1-20 alkylene are independently replaced with –O–, –NR1–, – C(=O)–, or optionally substituted heteroarylene. [0211] In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, optionally wherein one or more backbone carbon atoms in the optionally substituted linear C1-20 alkylene are independently replaced with –O–, –NR1–, –S–, –S(=O)–, – S(=O)2–, –C(=O)–, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, optionally wherein one or more backbone carbon atoms in the optionally substituted linear C1-20 alkylene are independently replaced with –O–, –NR1–, –C(=O)–, or optionally substituted heteroarylene.
U1197.70243WO00 66/193 #13587456v2 [0212] In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, wherein one or more backbone carbon atoms in the optionally substituted linear C1-20 alkylene are independently replaced with –O–, –NR1–, –C(=O)–, or optionally substituted heteroarylene. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, wherein at least 1, 2, 3, 4, or 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –O–. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, wherein at least 1 backbone carbon atom in the optionally substituted linear C1-20 alkylene is replaced with –O–. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, wherein at least 2 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –O–. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, wherein at least 3 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –O–. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, wherein at least 4 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –O–. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, wherein at least 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –O–. [0213] In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, wherein at least 1, 2, 3, 4, or 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –NR1–. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, wherein at least 1 backbone carbon atoms in the optionally substituted linear C1-20 alkylene is replaced with –NR1–. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, wherein at least 2 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –NR1–. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, wherein at least 3 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –NR1–. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, wherein at least 4 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –NR1–. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, wherein at least 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –NR1–. [0214] In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, wherein at least 1, 2, 3, 4, or 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –C(=O)–. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, wherein at least 1 backbone
U1197.70243WO00 67/193 #13587456v2 carbon atom in the optionally substituted linear C1-20 alkylene is replaced with –C(=O)–. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, wherein at least 2 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –C(=O)–. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, wherein at least 3 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –C(=O)–. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, wherein at least 4 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –C(=O)–. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, wherein at least 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with –C(=O)–. [0215] In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, wherein at least 1, 2, 3, 4, or 5 backbone carbon atoms in the optionally substituted linear C1-20 alkylene are replaced with optionally substituted heteroarylene. In some embodiments, each instance of L independently comprises optionally substituted linear C1-20 alkylene, wherein at least 1 backbone carbon atom in the optionally substituted linear C1-20 alkylene is replaced with optionally substituted heteroarylene. [0216] In some embodiments, each instance of L independently comprises , , 10; and each y is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, each instance of L independently comprises
U1197.70243WO00 68/193 #13587456v2 , , , , , , , , , , , , , , or 10; and each y is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, each instance of L [0217] In some embodiments, each instance of L independently comprises optionally substituted heteroarylene, optionally wherein one or more backbone carbon atoms in the optionally substituted heteroarylene are independently replaced with –O–, –NR1–, –S–, –S(=O)–, –S(=O)2–, –C(=O)–, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene. In some embodiments, each instance of L independently comprises optionally substituted heteroarylene, optionally wherein one or more backbone carbon atoms in the optionally substituted heteroarylene are independently replaced with –
U1197.70243WO00 69/193 #13587456v2 O–, –NR1–, –C(=O)–, or optionally substituted heteroarylene. [0218] In some embodiments, each instance of L independently comprises optionally substituted heteroarylene. In some embodiments, each instance of L independently comprises optionally substituted 5-6 membered heteroarylene. In some embodiments, each instance of L independently comprises optionally substituted 5 membered heteroarylene. In some embodiments, each instance of L independently comprises optionally substituted 6 membered heteroarylene. In some embodiments, each instance of L independently comprises optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring heteroatoms selected from O, N, and S. In some embodiments, each instance of L independently comprises optionally substituted 5 membered heteroarylene having 1, 2, or 3 ring heteroatoms selected from O, N, and S. In some embodiments, each instance of L independently comprises optionally substituted 6 membered heteroarylene having 1, 2, or 3 ring heteroatoms selected from O, N, and S. [0219] In some embodiments, each instance of L independently comprises optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring N atoms. In some embodiments, each instance of L independently comprises optionally substituted 5-6 membered heteroarylene having 1 ring N atom. In some embodiments, each instance of L independently comprises optionally substituted 5-6 membered heteroarylene having 2 ring N atoms. In some embodiments, each instance of L independently comprises optionally substituted 5-6 membered heteroarylene having 3 ring N atoms. In some embodiments, each instance of L independently comprises optionally substituted 5 membered heteroarylene having 1, 2, or 3 ring N atoms. In some embodiments, each instance of L independently comprises optionally substituted 5 membered heteroarylene having 1 ring N atom. In some embodiments, each instance of L independently comprises optionally substituted 5 membered heteroarylene having 2 ring N atoms. In some embodiments, each instance of L independently comprises optionally substituted 5 membered heteroarylene having 3 ring N atoms. In some embodiments, each instance of L independently comprises optionally substituted 6 membered heteroarylene having 1, 2, or 3 ring N atoms. In some embodiments, each instance of L independently comprises optionally substituted 6 membered heteroarylene having 1 ring N atom. In some embodiments, each instance of L independently comprises optionally substituted 6 membered heteroarylene having 2 ring N atoms. In some embodiments, each instance of L independently comprises optionally substituted 6 membered heteroarylene having 3 ring N atoms. [0220] In some embodiments, each instance of L independently comprises , , , . In some embodiments, each instance of L independently
U1197.70243WO00 70/193 #13587456v2 comprises . In some embodiments, each instance of L independently comprises . In some embodiments, each instance of L independently comprises . [0221] In some embodiments, each instance of L independently comprises [0222] As generally described herein, each instance of R1 is independently hydrogen or optionally substituted alkyl. [0223] In some embodiments, at least one instance of R1 is hydrogen. [0224] In some embodiments, at least one instance of R1 is optionally substituted alkyl. In some embodiments, at least one instance of R1 is optionally substituted C1-12 alkyl. In some embodiments, at least one instance of R1 is optionally substituted C1-6 alkyl. In some embodiments, at least one instance of R1 is optionally substituted C1-3 alkyl. In some embodiments, at least one instance of R1 is unsubstituted alkyl. In some embodiments, at least one instance of R1 is unsubstituted C1-12 alkyl. In some embodiments, at least one instance of R1 is unsubstituted C1-6 alkyl. In some embodiments, at least one instance of R1 is unsubstituted C1-3 alkyl. In some embodiments, at least one instance of R1 is
U1197.70243WO00 71/193 #13587456v2 unsubstituted linear alkyl. In some embodiments, at least one instance of R1 is unsubstituted linear C1- 12 alkyl. In some embodiments, at least one instance of R1 is unsubstituted linear C1-6 alkyl. In some embodiments, at least one instance of R1 is unsubstituted linear C1-3 alkyl. [0225] In some embodiments, each instance of R1 is independently hydrogen. [0226] In some embodiments, each instance of R1 is independently optionally substituted alkyl. In some embodiments, each instance of R1 is independently optionally substituted C1-12 alkyl. In some embodiments, each instance of R1 is independently optionally substituted C1-6 alkyl. In some embodiments, each instance of R1 is independently optionally substituted C1-3 alkyl. In some embodiments, each instance of R1 is independently unsubstituted alkyl. In some embodiments, each instance of R1 is independently unsubstituted C1-12 alkyl. In some embodiments, each instance of R1 is independently unsubstituted C1-6 alkyl. In some embodiments, each instance of R1 is independently unsubstituted C1-3 alkyl. In some embodiments, each instance of R1 is independently unsubstituted linear alkyl. In some embodiments, each instance of R1 is independently unsubstituted linear C1-12 alkyl. In some embodiments, each instance of R1 is independently unsubstituted linear C1-6 alkyl. In some embodiments, each instance of R1 is independently unsubstituted linear C1-3 alkyl. X [0227] As generally described herein, each instance of X is independently a conjugated agent. In some embodiments, the conjugated agent is a radical of an agent. In some embodiments, each instance of X is independently a radical of an agent. [0228] In some embodiments, the agent is a small molecule compound (e.g., a small molecule organic compound). In some embodiments, the conjugated agent is a radical of a small molecule compound (e.g., a small molecule organic compound). In some embodiments, each instance of X is independently a radical of a small molecule compound (e.g., a small molecule organic compound). In some embodiments, an agent is a small molecule compound having a molecular weight of less than 2,000 daltons (Da), less than 1,500 Da, less than 1,000 Da, or less than 500 Da. In some embodiments, an agent is a small-molecule compound having a molecular weight of between about 100 and about 2,000 Da. For example, in some embodiments, the small-molecule compound has a molecular weight of between about 100 and about 1,500 Da, between about 100 and about 1,000 Da, between about 100 and about 750 Da, between about 100 and about 500 Da, between about 500 and about 2,000 Da, between about 500 and about 1,500 Da, or between about 500 and about 1,000 Da. [0229] In some embodiments, the conjugated agent is a radical of an agent selected from:
U1197.70243WO00 72/193 #13587456v2 or a pharmaceutically acceptable salt thereof. [0230] In some embodiments, the conjugated agent is a radical of LLL12-Hydrazone (18): Hydrazone, 18),
U1197.70243WO00 73/193 #13587456v2 or a pharmaceutically acceptable salt thereof. [0231] LLL12 refers to 5-hydroxy-9,10-dioxo-9,10-dihydroanthracene-1-sulfonamide (CAS No. 1260247-42-4), or a pharmaceutically acceptable salt thereof, which is a signal transducer and activator of transcription 3 (STAT3) inhibitor. Imiquimod refers to 1-isobutyl-1H-imidazo[4,5- c]quinolin-4-amine (CAS No.99011-02-6), or a pharmaceutically acceptable salt thereof, which is a Toll-like receptor 7 (TLR7) agonist. NLG-919 (IDO-IN-7) refers to 1-cyclohexyl-2-(5H-imidazo[5,1- a]isoindol-5-yl)ethan-1-ol (CAS No.1402836-58-1), or a pharmaceutically acceptable salt thereof, which is an indoleamine 2,3-dioxygenase (IDO) inhibitor. L-NMMA refers to N5- [imino(methylamino)methyl]-L-ornithine (L-NG-monomethyl Arginine) (CAS No.53308-83-1, acetate salt), or a pharmaceutically acceptable salt thereof, which is a nitric oxide synthase (NOS) inhibitor. Nor-NOHA refers to (S)-2-amino-4-(3-hydroxyguanidino)butanoic acid (Nω-Hydroxy-nor- L-arginine) (CAS No.291758-32-2, dichloride salt), or a pharmaceutically acceptable salt thereof, which is an arginase inhibitor. CA-170 refers to (((S)-3-amino-1-(3-((R)-1-amino-2-hydroxyethyl)- 1,2,4-oxadiazol-5-yl)-3-oxopropyl)carbamoyl)-L-threonine (CAS No.1673534-76-3), or a pharmaceutically acceptable salt thereof, which is a programmed death-ligand 1 (PD-L1) inhibitor. Triciribine phosphate refers to ((2S,3R,4S,5S)-5-(3-amino-5-methyl-1,4,5,6,8-pentaazaacenaphthylen- 1(5H)-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl dihydrogen phosphate (CAS No.61966-08-3), or a pharmaceutically acceptable salt thereof, which is a protein kinase B inhibitor. AB-680 refers to (((((2R,3S,4R,5R)-5-(6-chloro-4-(((S)-1-(2-fluorophenyl)ethyl)amino)-1H-pyrazolo[3,4-b]pyridin-1- yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid (CAS No.2105904-82-1), or a pharmaceutically acceptable salt thereof, which is a cluster of differentiation 73 (CD73) inhibitor.2′,3′-cGAMP (2′,3′-cyclic GMP-AMP) refers to 2-amino-9- ((5R,7R,8R,12aR,14R,15R,15aS,16R)-14-(6-amino-9H-purin-9-yl)-2,10,15,16-tetrahydroxy-2,10- dioxidooctahydro-12H-5,8-methanofuro[3,2-l][1,3,6,9,11]pentaoxa[2,10]diphosphacyclotetradecin-7- yl)-3,9-dihydro-6H-purin-6-one (CAS No.1441190-66-4), or a pharmaceutically acceptable salt thereof, which is a stimulator of interferon genes (STING) agonist. [0232] In some embodiments, the conjugated agent is a radical of an agent selected from LLL12, imiquimod, and NLG-919. In some embodiments, the conjugated agent is a radical of LLL12. In some embodiments, the conjugated agent is a radical of imiquimod. In some embodiments, the conjugated agent is a radical of NLG-919. In some embodiments, the conjugated agent is a radical of an agent selected from L-NMMA, nor-NOHA, and CA-170. In some embodiments, the conjugated agent is a radical of L-NMMA. In some embodiments, the conjugated agent is a radical of nor-NOHA. In some embodiments, the conjugated agent is a radical of CA-170. In some embodiments, the conjugated agent is a radical of an agent selected from triciribine phosphate, AB-680, and 2′,3′-cGAMP. In some embodiments, the conjugated agent is a radical of triciribine phosphate. In some embodiments, the conjugated agent is a radical of AB-680. In some embodiments, the conjugated agent is a radical of
U1197.70243WO00 74/193 #13587456v2 2′,3′-cGAMP. In some embodiments, the conjugated agent is a radical of an agent selected from LLL12 and LLL12-Hydrazone. [0233] In some embodiments, at least one instance of X is a radical of an agent selected from LLL12, imiquimod, NLG-919, L-NMMA, nor-NOHA, CA-170, triciribine phosphate, AB-680, and 2′,3′- cGAMP. In some embodiments, at least one instance of X is a radical of an agent selected from LLL12, imiquimod, and NLG-919. In some embodiments, at least one instance of X is a radical of LLL12. In some embodiments, at least one instance of X is a radical of imiquimod. In some embodiments, at least one instance of X is a radical of NLG-919. In some embodiments, at least one instance of X is a radical of an agent selected from L-NMMA, nor-NOHA, and CA-170. In some embodiments, at least one instance of X is a radical of L-NMMA. In some embodiments, at least one instance of X is a radical of nor-NOHA. In some embodiments, at least one instance of X is a radical of CA-170. In some embodiments, at least one instance of X is a radical of an agent selected from triciribine phosphate, AB-680, and 2′,3′-cGAMP. In some embodiments, at least one instance of X is a radical of triciribine phosphate. In some embodiments, at least one instance of X is a radical of AB- 680. In some embodiments, at least one instance of X is a radical of 2′,3′-cGAMP. [0234] In some embodiments, at least one instance of X is a radical of an agent selected from LLL12, imiquimod, NLG-919, L-NMMA, nor-NOHA, CA-170, triciribine phosphate, AB-680, 2′,3′-cGAMP, and LLL12-Hydrazone. In some embodiments, at least one instance of X is a radical of an agent selected from LLL12 and LLL12-Hydrazone. In some embodiments, at least one instance of X is a radical of LLL12-Hydrazone. [0235] In some embodiments, each instance of X is independently a radical of an agent selected from LLL12, imiquimod, NLG-919, L-NMMA, nor-NOHA, CA-170, triciribine phosphate, AB-680, and 2′,3′-cGAMP. In some embodiments, each instance of X is independently a radical of an agent selected from LLL12, imiquimod, and NLG-919. In some embodiments, each instance of X is independently a radical of LLL12. In some embodiments, each instance of X is independently a radical of imiquimod. In some embodiments, each instance of X is independently a radical of NLG- 919. In some embodiments, each instance of X is independently a radical of an agent selected from L- NMMA, nor-NOHA, and CA-170. In some embodiments, each instance of X is independently a radical of L-NMMA. In some embodiments, each instance of X is independently a radical of nor- NOHA. In some embodiments, each instance of X is independently a radical of CA-170. In some embodiments, each instance of X is independently a radical of an agent selected from triciribine phosphate, AB-680, and 2′,3′-cGAMP. In some embodiments, each instance of X is independently a radical of triciribine phosphate. In some embodiments, each instance of X is independently a radical of AB-680. In some embodiments, each instance of X is independently a radical of 2′,3′-cGAMP. [0236] In some embodiments, each instance of X is independently a radical of an agent selected from LLL12, imiquimod, NLG-919, L-NMMA, nor-NOHA, CA-170, triciribine phosphate, AB-680, 2′,3′- cGAMP, and LLL12-Hydrazone. In some embodiments, each instance of X is independently a radical
U1197.70243WO00 75/193 #13587456v2 of an agent selected from LLL12 and LLL12-Hydrazone. In some embodiments, each instance of X is independently a radical of LLL12-Hydrazone. [0237] In some embodiments, the agent is a hydrophobic agent, a cationic agent, or an anionic agent. In some embodiments, the agent is a hydrophobic agent. In some embodiments, the hydrophobic agent has a logP value of greater than 0. In some embodiments, the hydrophobic agent is selected from LLL12, imiquimod, and NLG-919, and pharmaceutically acceptable salts thereof. [0238] In some embodiments, the agent is a cationic agent. In some embodiments, the cationic agent is positively charged at physiological pH. In some embodiments, the cationic agent is selected from L- NMMA, nor-NOHA, and CA-170, and pharmaceutically acceptable salts thereof. [0239] In some embodiments, the agent is an anionic agent. In some embodiments, the anionic agent is negatively charged at physiological pH. In some embodiments, the anionic agent is selected from triciribine phosphate, AB-680, and 2′,3′-cGAMP, and pharmaceutically acceptable salts thereof. [0240] In some embodiments, the agent is a STAT3 inhibitor, a TLR7 agonist, an IDO inhibitor, an NOS inhibitor, an arginase inhibitor, a PD-L1 inhibitor, a protein kinase B inhibitor, a CD73 inhibitor, or a STING agonist. In some embodiments, the agent is a STAT3 inhibitor, a TLR7 agonist, or an IDO inhibitor. In some embodiments, the agent is a STAT3 inhibitor. In some embodiments, the agent is a TLR7 agonist. In some embodiments, the agent is an IDO inhibitor. In some embodiments, the agent is an NOS inhibitor, an arginase inhibitor, a PD-L1 inhibitor. In some embodiments, the agent is an NOS inhibitor. In some embodiments, the agent is an arginase inhibitor. In some embodiments, the agent is a PD-L1 inhibitor. In some embodiments, the agent is a protein kinase B inhibitor, a CD73 inhibitor, or a STING agonist. In some embodiments, the agent is a protein kinase B inhibitor. In some embodiments, the agent is a CD73 inhibitor. In some embodiments, the agent is a STING agonist. [0241] In some embodiments, the STAT3 inhibitor is LLL12. In some embodiments, the STAT3 inhibitor is LLL12-Hydrazone. In some embodiments, the TLR7 agonist is imiquimod. In some embodiments, the IDO inhibitor is NLG-919. In some embodiments, the NOS inhibitor is L-NMMA. In some embodiments, the arginase inhibitor is nor-NOHA. In some embodiments, the PD-L1 inhibitor is CA-170. In some embodiments, the protein kinase B inhibitor is triciribine phosphate. In some embodiments, the CD73 inhibitor is AB-680. In some embodiments, the STING agonist is 2′,3′- cGAMP.
U1197.70243WO00 76/193 #13587456v2 [0242] In some embodiments, at least one instance of X is selected from formulae (ii), (iii), (iv), (v), (vi), (vii), (viii)
U1197.70243WO00 77/193 #13587456v2 [0243] In some embodiments, at least one instance of X is selected from formulae (ii), (iii), (iv), (v), (vi), (vii), (viii) [0244] In some embodiments, at least one instance of X is selected from formulae (ii), (iii), and (iv). In some embodiments, at least one instance of X is of formula (ii). In some embodiments, at least one instance of X is of formula (iii). In some embodiments, at least one instance of X is of formula (iv). In some embodiments, at least one instance of X is selected from formulae (v), (vi), and (vii). In some embodiments, at least one instance of X is of formula (v). In some embodiments, at least one instance of X is of formula (vi). In some embodiments, at least one instance of X is of formula (vii). In some embodiments, at least one instance of X is selected from formulae (viii), (ix), and (x). In some embodiments, at least one instance of X is of formula (viii). In some embodiments, at least one
U1197.70243WO00 78/193 #13587456v2 instance of X is of formula (ix). In some embodiments, at least one instance of X is of formula (x). In some embodiments, at least one instance of X is selected from formulae (ii) and (xi). In some embodiments, at least one instance of X is of formula (xi). [0245] In some embodiments, each instance of X is independently selected from formulae (ii), (iii), (iv), (v), (vi), (vii), (viii), (ix), and (x). In some embodiments, each instance of X is independently selected from formulae (ii), (iii), and (iv). In some embodiments, each instance of X is independently of formula (ii). In some embodiments, each instance of X is independently of formula (iii). In some embodiments, each instance of X is independently of formula (iv). In some embodiments, each instance of X is independently selected from formulae (v), (vi), and (vii). In some embodiments, each instance of X is independently of formula (v). In some embodiments, each instance of X is independently of formula (vi). In some embodiments, each instance of X is independently of formula (vii). In some embodiments, each instance of X is independently selected from formulae (viii), (ix), and (x). In some embodiments, each instance of X is independently of formula (viii). In some embodiments, each instance of X is independently of formula (ix). In some embodiments, each instance of X is independently of formula (x). In some embodiments, each instance of X is independently selected from formulae (ii) and (xi). In some embodiments, each instance of X is independently of formula (xi). [0246] In some embodiments, the compound of Formula (I) or Formula (I′) is selected from
U1197.70243WO00 79/193 #13587456v2
or a pharmaceutically acceptable salt thereof. [0247] In some embodiments, the compound of Formula (I) or Formula (I′) is selected from Formulae
U1197.70243WO00 80/193 #13587456v2 or a pharmaceutically acceptable salt thereof. [0248] In some embodiments, the compound of Formula (I) or Formula (I′) is selected from Formulae (II), (III), (IV), and (XI). In some embodiments, the compound of Formula (I) or Formula (I′) is selected from Formulae (II), (III), and (IV). In some embodiments, the compound of Formula (I) or Formula (I′) is of Formula (II). In some embodiments, the compound of Formula (I) or Formula (I′) is of Formula (III). In some embodiments, the compound of Formula (I) or Formula (I′) is of Formula (IV). In some embodiments, the compound of Formula (I) or Formula (I′) is selected from Formulae (V), (VI), and (VII). In some embodiments, the compound of Formula (I) or Formula (I′) is
U1197.70243WO00 81/193 #13587456v2 of Formula (V). In some embodiments, the compound of Formula (I) or Formula (I′) is of Formula (VI). In some embodiments, the compound of Formula (I) or Formula (I′) is of Formula (VII). In some embodiments, the compound of Formula (I) or Formula (I′) is selected from Formulae (VIII), (IX), and (X). In some embodiments, the compound of Formula (I) or Formula (I′) is of Formula (VIII). In some embodiments, the compound of Formula (I) or Formula (I′) is of Formula (IX). In some embodiments, the compound of Formula (I) or Formula (I′) is of Formula (X). In some embodiments, the compound of Formula (I) or Formula (I′) is selected from Formulae (II) and (XI). In some embodiments, the compound of Formula (I) or Formula (I′) is of Formula (XI). [0249] In some embodiments, the compound of Formula (I) is selected from Formulae (II), (III), (IV), and (XI). In some embodiments, the compound of Formula (I) is selected from Formulae (II), (III), and (IV). In some embodiments, the compound of Formula (I) is of Formula (II). In some embodiments, the compound of Formula (I) is of Formula (III). In some embodiments, the compound of Formula (I) is of Formula (IV). In some embodiments, the compound of Formula (I) is selected from Formulae (V), (VI), and (VII). In some embodiments, the compound of Formula (I) is of Formula (V). In some embodiments, the compound of Formula (I) is of Formula (VI). In some embodiments, the compound of Formula (I) is of Formula (VII). In some embodiments, the compound of Formula (I) is selected from Formulae (VIII), (IX), and (X). In some embodiments, the compound of Formula (I) is of Formula (VIII). In some embodiments, the compound of Formula (I) is of Formula (IX). In some embodiments, the compound of Formula (I) is of Formula (X). In some embodiments, the compound of Formula (I) is selected from Formulae (II) and (XI). In some embodiments, the compound of Formula (I) is of Formula (XI). [0250] In some embodiments, the compound of Formula (I′) is selected from Formulae (II), (III), (IV), and (XI). In some embodiments, the compound of Formula (I′) is selected from Formulae (II), (III), and (IV). In some embodiments, the compound of Formula (I′) is of Formula (II). In some embodiments, the compound of Formula (I′) is of Formula (III). In some embodiments, the compound of Formula (I′) is of Formula (IV). In some embodiments, the compound of Formula (I′) is selected from Formulae (V), (VI), and (VII). In some embodiments, the compound of Formula (I′) is of Formula (V). In some embodiments, the compound of Formula (I′) is of Formula (VI). In some embodiments, the compound of Formula (I′) is of Formula (VII). In some embodiments, the compound of Formula (I′) is selected from Formulae (VIII), (IX), and (X). In some embodiments, the compound of Formula (I′) is of Formula (VIII). In some embodiments, the compound of Formula (I′) is of Formula (IX). In some embodiments, the compound of Formula (I′) is of Formula (X). In some embodiments, the compound of Formula (I′) is selected from Formulae (II) and (XI). In some embodiments, the compound of Formula (I′) is of Formula (XI). [0251] In some embodiments, the compound of Formula (I) can be characterized in terms of mass percentage (e.g., % by mass (m/m)) of X (e.g., of conjugated agent). In some embodiments, mass percentage refers to a molecular weight (Da) percentage of X (e.g., of conjugated agent) in the
U1197.70243WO00 82/193 #13587456v2 compound of Formula (I). In some embodiments, mass percentage can be determined by the general formula of: (X (e.g., conjugated agent) MW) / (compound of Formula (I) MW) × 100. For example, in some embodiments, (X (e.g., conjugated agent) MW) can be determined by calculating or approximating the molecular weight of X (e.g., of a conjugated agent) as a single molecule or compound (conjugated or unconjugated), and multiplying this value by the number of terminal sites at which X (e.g., of conjugated agent) is present in the compound of Formula (I) (e.g., multiplying by p). In some embodiments, (X (e.g., conjugated agent) MW) can be determined by calculating or approximating the sum of the atomic mass of all atoms which form X (e.g., conjugated agent) in the compound of Formula (I). The value for (X (e.g., conjugated agent) MW) can be taken as a fraction of total molecular weight of the compound of Formula (I) (compound of Formula (I) MW), and multiplied by 100 to provide a mass percentage. In some embodiments, mass percentage can be determined by experimental or empirical means. For example, in some embodiments, mass percentage can be determined using proton nuclear magnetic resonance (1H NMR) or other analytical methods known in the art. [0252] In some embodiments, the compound of Formula (I′) can be characterized in terms of mass percentage (e.g., % by mass (m/m)) of X. In some embodiments, mass percentage refers to a molecular weight (Da) percentage of X in the compound of Formula (I′). In some embodiments, mass percentage can be determined by the general formula of: (X MW) / (compound of Formula (I′) MW) × 100. For example, in some embodiments, (X MW) can be determined by calculating or approximating the molecular weight of X as a single molecule or compound (conjugated or unconjugated), and multiplying this value by the number of terminal sites at which X is present in the compound of Formula (I′) (e.g., multiplying by p). In some embodiments, (X MW) can be determined by calculating or approximating the sum of the atomic mass of all atoms which form X in the compound of Formula (I′). The value for (X MW) can be taken as a fraction of total molecular weight of the compound of Formula (I′) (compound of Formula (I′) MW), and multiplied by 100 to provide a mass percentage. In some embodiments, mass percentage can be determined by experimental or empirical means. For example, in some embodiments, mass percentage can be determined using proton nuclear magnetic resonance (1H NMR) or other analytical methods known in the art. [0253] In some embodiments, the compound of Formula (I) comprises between about 1% and about 20%, about 1% and about 15%, about 1% and about 14%, about 1% and about 13%, about 1% and about 12%, about 1% and about 11%, about 1% and about 10%, about 1% and about 9%, about 1% and about 8%, about 1% and about 7%, about 1% and about 6%, about 1% and about 5%, about 2% and about 15%, about 2% and about 10%, about 3% and about 15%, about 3% and about 10%, about 4% and about 15%, about 5% and about 10%, about 6% and about 15%, about 7% and about 10%, about 8% and about 15%, or about 9% and about 10% by mass (m/m) of X (e.g., of conjugated agent). In some embodiments, the compound of Formula (I) comprises between about 1% and about 20% by mass of X (e.g., of conjugated agent). In some embodiments, the compound of Formula (I) comprises
U1197.70243WO00 83/193 #13587456v2 between about 1% and about 10% by mass of X (e.g., of conjugated agent). In some embodiments, the compound of Formula (I) comprises between about 5% and about 10% by mass of X (e.g., of conjugated agent). [0254] In some embodiments, the compound of Formula (I′) comprises between about 1% and about 20%, about 1% and about 15%, about 1% and about 14%, about 1% and about 13%, about 1% and about 12%, about 1% and about 11%, about 1% and about 10%, about 1% and about 9%, about 1% and about 8%, about 1% and about 7%, about 1% and about 6%, about 1% and about 5%, about 2% and about 15%, about 2% and about 10%, about 3% and about 15%, about 3% and about 10%, about 4% and about 15%, about 5% and about 10%, about 6% and about 15%, about 7% and about 10%, about 8% and about 15%, or about 9% and about 10% by mass (m/m) of X. In some embodiments, the compound of Formula (I′) comprises between about 1% and about 20% by mass of X. In some embodiments, the compound of Formula (I′) comprises between about 1% and about 10% by mass of X. In some embodiments, the compound of Formula (I′) comprises between about 5% and about 10% by mass of X. [0255] In some embodiments, the compound of Formula (II) comprises between about 1% and about 20%, about 1% and about 15%, about 1% and about 14%, about 1% and about 13%, about 1% and about 12%, about 1% and about 11%, about 1% and about 10%, about 1% and about 9%, about 1% and about 8%, about 1% and about 7%, about 1% and about 6%, about 1% and about 5%, about 2% and about 15%, about 2% and about 10%, about 3% and about 15%, about 3% and about 10%, about 4% and about 15%, about 5% and about 10%, about 6% and about 15%, about 7% and about 10%, about 8% and about 15%, or about 9% and about 10% by mass (m/m) of formula (ii). In some embodiments, the compound of Formula (II) comprises between about 1% and about 20% by mass of formula (ii). In some embodiments, the compound of Formula (II) comprises between about 1% and about 10% by mass of formula (ii). In some embodiments, the compound of Formula (II) comprises between about 5% and about 10% by mass of formula (ii). [0256] In some embodiments, the compound of Formula (III) comprises between about 1% and about 20%, about 1% and about 15%, about 1% and about 14%, about 1% and about 13%, about 1% and about 12%, about 1% and about 11%, about 1% and about 10%, about 1% and about 9%, about 1% and about 8%, about 1% and about 7%, about 1% and about 6%, about 1% and about 5%, about 2% and about 15%, about 2% and about 10%, about 3% and about 15%, about 3% and about 10%, about 4% and about 15%, about 5% and about 10%, about 6% and about 15%, about 7% and about 10%, about 8% and about 15%, or about 9% and about 10% by mass (m/m) of formula (iii). In some embodiments, the compound of Formula (III) comprises between about 1% and about 20% by mass of formula (iii). In some embodiments, the compound of Formula (III) comprises between about 1% and about 10% by mass of formula (iii). In some embodiments, the compound of Formula (III) comprises between about 5% and about 10% by mass of formula (iii).
U1197.70243WO00 84/193 #13587456v2 [0257] In some embodiments, the compound of Formula (IV) comprises between about 1% and about 20%, about 1% and about 15%, about 1% and about 14%, about 1% and about 13%, about 1% and about 12%, about 1% and about 11%, about 1% and about 10%, about 1% and about 9%, about 1% and about 8%, about 1% and about 7%, about 1% and about 6%, about 1% and about 5%, about 2% and about 15%, about 2% and about 10%, about 3% and about 15%, about 3% and about 10%, about 4% and about 15%, about 5% and about 10%, about 6% and about 15%, about 7% and about 10%, about 8% and about 15%, or about 9% and about 10% by mass (m/m) of formula (iv). In some embodiments, the compound of Formula (IV) comprises between about 1% and about 20% by mass of formula (iv). In some embodiments, the compound of Formula (IV) comprises between about 1% and about 10% by mass of formula (iv). In some embodiments, the compound of Formula (IV) comprises between about 5% and about 10% by mass of formula (iv). [0258] In some embodiments, the compound of Formula (V) comprises between about 1% and about 20%, about 1% and about 15%, about 1% and about 14%, about 1% and about 13%, about 1% and about 12%, about 1% and about 11%, about 1% and about 10%, about 1% and about 9%, about 1% and about 8%, about 1% and about 7%, about 1% and about 6%, about 1% and about 5%, about 2% and about 15%, about 2% and about 10%, about 3% and about 15%, about 3% and about 10%, about 4% and about 15%, about 5% and about 10%, about 6% and about 15%, about 7% and about 10%, about 8% and about 15%, or about 9% and about 10% by mass (m/m) of formula (v). In some embodiments, the compound of Formula (V) comprises between about 1% and about 20% by mass of formula (v). In some embodiments, the compound of Formula (V) comprises between about 1% and about 10% by mass of formula (v). In some embodiments, the compound of Formula (V) comprises between about 5% and about 10% by mass of formula (v). [0259] In some embodiments, the compound of Formula (VI) comprises between about 1% and about 20%, about 1% and about 15%, about 1% and about 14%, about 1% and about 13%, about 1% and about 12%, about 1% and about 11%, about 1% and about 10%, about 1% and about 9%, about 1% and about 8%, about 1% and about 7%, about 1% and about 6%, about 1% and about 5%, about 2% and about 15%, about 2% and about 10%, about 3% and about 15%, about 3% and about 10%, about 4% and about 15%, about 5% and about 10%, about 6% and about 15%, about 7% and about 10%, about 8% and about 15%, or about 9% and about 10% by mass (m/m) of formula (vi). In some embodiments, the compound of Formula (VI) comprises between about 1% and about 20% by mass of formula (vi). In some embodiments, the compound of Formula (VI) comprises between about 1% and about 10% by mass of formula (vi). In some embodiments, the compound of Formula (VI) comprises between about 5% and about 10% by mass of formula (vi). [0260] In some embodiments, the compound of Formula (VII) comprises between about 1% and about 20%, about 1% and about 15%, about 1% and about 14%, about 1% and about 13%, about 1% and about 12%, about 1% and about 11%, about 1% and about 10%, about 1% and about 9%, about 1% and about 8%, about 1% and about 7%, about 1% and about 6%, about 1% and about 5%, about
U1197.70243WO00 85/193 #13587456v2 2% and about 15%, about 2% and about 10%, about 3% and about 15%, about 3% and about 10%, about 4% and about 15%, about 5% and about 10%, about 6% and about 15%, about 7% and about 10%, about 8% and about 15%, or about 9% and about 10% by mass (m/m) of formula (vii). In some embodiments, the compound of Formula (VII) comprises between about 1% and about 20% by mass of formula (vii). In some embodiments, the compound of Formula (VII) comprises between about 1% and about 10% by mass of formula (vii). In some embodiments, the compound of Formula (VII) comprises between about 5% and about 10% by mass of formula (vii). [0261] In some embodiments, the compound of Formula (VIII) comprises between about 1% and about 20%, about 1% and about 15%, about 1% and about 14%, about 1% and about 13%, about 1% and about 12%, about 1% and about 11%, about 1% and about 10%, about 1% and about 9%, about 1% and about 8%, about 1% and about 7%, about 1% and about 6%, about 1% and about 5%, about 2% and about 15%, about 2% and about 10%, about 3% and about 15%, about 3% and about 10%, about 4% and about 15%, about 5% and about 10%, about 6% and about 15%, about 7% and about 10%, about 8% and about 15%, or about 9% and about 10% by mass (m/m) of formula (viii). In some embodiments, the compound of Formula (VIII) comprises between about 1% and about 20% by mass of formula (viii). In some embodiments, the compound of Formula (VIII) comprises between about 1% and about 10% by mass of formula (viii). In some embodiments, the compound of Formula (VIII) comprises between about 5% and about 10% by mass of formula (viii). [0262] In some embodiments, the compound of Formula (IX) comprises between about 1% and about 20%, about 1% and about 15%, about 1% and about 14%, about 1% and about 13%, about 1% and about 12%, about 1% and about 11%, about 1% and about 10%, about 1% and about 9%, about 1% and about 8%, about 1% and about 7%, about 1% and about 6%, about 1% and about 5%, about 2% and about 15%, about 2% and about 10%, about 3% and about 15%, about 3% and about 10%, about 4% and about 15%, about 5% and about 10%, about 6% and about 15%, about 7% and about 10%, about 8% and about 15%, or about 9% and about 10% by mass (m/m) of formula (ix). In some embodiments, the compound of Formula (IX) comprises between about 1% and about 20% by mass of formula (ix). In some embodiments, the compound of Formula (IX) comprises between about 1% and about 10% by mass of formula (ix). In some embodiments, the compound of Formula (IX) comprises between about 5% and about 10% by mass of formula (ix). [0263] In some embodiments, the compound of Formula (X) comprises between about 1% and about 20%, about 1% and about 15%, about 1% and about 14%, about 1% and about 13%, about 1% and about 12%, about 1% and about 11%, about 1% and about 10%, about 1% and about 9%, about 1% and about 8%, about 1% and about 7%, about 1% and about 6%, about 1% and about 5%, about 2% and about 15%, about 2% and about 10%, about 3% and about 15%, about 3% and about 10%, about 4% and about 15%, about 5% and about 10%, about 6% and about 15%, about 7% and about 10%, about 8% and about 15%, or about 9% and about 10% by mass (m/m) of formula (x). In some embodiments, the compound of Formula (X) comprises between about 1% and about 20% by mass of
U1197.70243WO00 86/193 #13587456v2 formula (x). In some embodiments, the compound of Formula (X) comprises between about 1% and about 10% by mass of formula (x). In some embodiments, the compound of Formula (X) comprises between about 5% and about 10% by mass of formula (x). [0264] In some embodiments, the compound of Formula (XI) comprises between about 1% and about 20%, about 1% and about 15%, about 1% and about 14%, about 1% and about 13%, about 1% and about 12%, about 1% and about 11%, about 1% and about 10%, about 1% and about 9%, about 1% and about 8%, about 1% and about 7%, about 1% and about 6%, about 1% and about 5%, about 2% and about 15%, about 2% and about 10%, about 3% and about 15%, about 3% and about 10%, about 4% and about 15%, about 5% and about 10%, about 6% and about 15%, about 7% and about 10%, about 8% and about 15%, or about 9% and about 10% by mass (m/m) of formula (xi). In some embodiments, the compound of Formula (XI) comprises between about 1% and about 20% by mass of formula (xi). In some embodiments, the compound of Formula (XI) comprises between about 1% and about 10% by mass of formula (xi). In some embodiments, the compound of Formula (XI) comprises between about 5% and about 10% by mass of formula (xi). Y and RA [0265] As generally described herein, each instance of Y is independently –ORA, –N(RA)2, or –C(=O)ORA. [0266] In some embodiments, at least one instance of Y is –ORA. In some embodiments, each instance of Y is independently –ORA. [0267] In some embodiments, at least one instance of Y is –OH. In some embodiments, each instance of Y is independently –OH. [0268] In some embodiments, at least one instance of Y is –ORA, wherein RA is hydrogen, optionally substituted alkyl, optionally substituted alkynyl, or optionally substituted heteroalkyl. In some embodiments, at least one instance of Y is –ORA, wherein RA is hydrogen or optionally substituted alkyl. In some embodiments, at least one instance of Y is –ORA, wherein RA is hydrogen or optionally substituted alkynyl. In some embodiments, at least one instance of Y is –ORA, wherein RA is hydrogen or optionally substituted heteroalkyl. In some embodiments, at least one instance of Y is – ORA, wherein RA is hydrogen, optionally substituted alkyl, or optionally substituted alkynyl. In some embodiments, at least one instance of Y is –ORA, wherein RA is hydrogen, optionally substituted alkyl, or optionally substituted heteroalkyl. In some embodiments, at least one instance of Y is –ORA, wherein RA is hydrogen, optionally substituted alkynyl, or optionally substituted heteroalkyl. [0269] In some embodiments, each instance of Y is independently –ORA, wherein RA is hydrogen, optionally substituted alkyl, optionally substituted alkynyl, or optionally substituted heteroalkyl. In some embodiments, each instance of Y is independently –ORA, wherein RA is hydrogen or optionally substituted alkyl. In some embodiments, each instance of Y is independently –ORA, wherein RA is hydrogen or optionally substituted alkynyl. In some embodiments, each instance of Y is
U1197.70243WO00 87/193 #13587456v2 independently –ORA, wherein RA is hydrogen or optionally substituted heteroalkyl. In some embodiments, each instance of Y is independently –ORA, wherein RA is hydrogen, optionally substituted alkyl, or optionally substituted alkynyl. In some embodiments, each instance of Y is independently –ORA, wherein RA is hydrogen, optionally substituted alkyl, or optionally substituted heteroalkyl. In some embodiments, each instance of Y is independently –ORA, wherein RA is hydrogen, optionally substituted alkynyl, or optionally substituted heteroalkyl. [0270] In some embodiments, at least one instance of Y is –ORA, wherein RA is optionally substituted alkyl, optionally substituted alkynyl, or optionally substituted heteroalkyl. In some embodiments, at least one instance of Y is –ORA, wherein RA is optionally substituted alkyl or optionally substituted alkynyl. In some embodiments, at least one instance of Y is –ORA, wherein RA is optionally substituted alkyl or optionally substituted heteroalkyl. In some embodiments, at least one instance of Y is –ORA, wherein RA is optionally substituted alkynyl or optionally substituted heteroalkyl. [0271] In some embodiments, at least one instance of Y is –ORA, wherein RA is hydrogen, optionally substituted C1-10 alkyl, optionally substituted C1-10 alkynyl, or optionally substituted C1-10 heteroalkyl. In some embodiments, at least one instance of Y is –ORA, wherein RA is hydrogen, optionally substituted linear C1-10 alkyl, optionally substituted linear C1-10 alkynyl, or optionally substituted linear C1-10 heteroalkyl. In some embodiments, at least one instance of Y is –ORA, wherein RA is hydrogen, optionally substituted C1-6 alkyl, optionally substituted C1-6 alkynyl, or optionally substituted C1-6 heteroalkyl. In some embodiments, at least one instance of Y is –ORA, wherein RA is hydrogen, optionally substituted linear C1-6 alkyl, optionally substituted linear C1-6 alkynyl, or optionally substituted linear C1-6 heteroalkyl. [0272] In some embodiments, at least one instance of Y is –ORA, wherein RA is hydrogen, C1-10 alkyl, C1-10 alkynyl, or C1-10 heteroalkyl, wherein the alkyl, alkynyl, or heteroalkyl is substituted with an oxo group (=O) and/or an amino group (–NH2). In some embodiments, at least one instance of Y is –ORA, wherein RA is hydrogen, C1-10 alkyl, C1-10 alkynyl, or C1-10 heteroalkyl, wherein the alkyl, alkynyl, or heteroalkyl is substituted with an oxo group (=O). In some embodiments, at least one instance of Y is –ORA, wherein RA is hydrogen, linear C1-10 alkyl, linear C1-10 alkynyl, or linear C1-10 heteroalkyl, wherein the alkyl, alkynyl, or heteroalkyl is substituted with an oxo group (=O) and/or an amino group (–NH2). In some embodiments, at least one instance of Y is –ORA, wherein RA is hydrogen, linear C1-10 alkyl, linear C1-10 alkynyl, or linear C1-10 heteroalkyl, wherein the alkyl, alkynyl, or heteroalkyl is substituted with an oxo group (=O). In some embodiments, at least one instance of Y is –ORA, wherein RA is hydrogen, C1-6 alkyl, C1-6 alkynyl, or C1-6 heteroalkyl, wherein the alkyl, alkynyl, or heteroalkyl is substituted with an oxo group (=O) and/or an amino group (–NH2). In some embodiments, at least one instance of Y is –ORA, wherein RA is hydrogen, C1-6 alkyl, C1-6 alkynyl, or C1-6 heteroalkyl, wherein the alkyl, alkynyl, or heteroalkyl is substituted with an oxo group (=O). In some embodiments, at least one instance of Y is –ORA, wherein RA is hydrogen, linear C1-6 alkyl,
U1197.70243WO00 88/193 #13587456v2 linear C1-6 alkynyl, or linear C1-6 heteroalkyl, wherein the alkyl, alkynyl, or heteroalkyl is substituted with an oxo group (=O) and/or an amino group (–NH2). In some embodiments, at least one instance of Y is –ORA, wherein RA is hydrogen, linear C1-6 alkyl, linear C1-6 alkynyl, or linear C1-6 heteroalkyl, wherein the alkyl, alkynyl, or heteroalkyl is substituted with an oxo group (=O). [0273] In some embodiments, each instance of Y is independently –ORA, wherein RA is hydrogen, optionally substituted C1-10 alkyl, optionally substituted C1-10 alkynyl, or optionally substituted C1-10 heteroalkyl. In some embodiments, each instance of Y is independently –ORA, wherein RA is hydrogen, optionally substituted linear C1-10 alkyl, optionally substituted linear C1-10 alkynyl, or optionally substituted linear C1-10 heteroalkyl. In some embodiments, each instance of Y is independently –ORA, wherein RA is hydrogen, optionally substituted C1-6 alkyl, optionally substituted C1-6 alkynyl, or optionally substituted C1-6 heteroalkyl. In some embodiments, each instance of Y is independently –ORA, wherein RA is hydrogen, optionally substituted linear C1-6 alkyl, optionally substituted linear C1-6 alkynyl, or optionally substituted linear C1-6 heteroalkyl. [0274] In some embodiments, each instance of Y is independently –ORA, wherein RA is hydrogen, C1-10 alkyl, C1-10 alkynyl, or C1-10 heteroalkyl, wherein the alkyl, alkynyl, or heteroalkyl is substituted with an oxo group (=O) and/or an amino group (–NH2). In some embodiments, each instance of Y is independently –ORA, wherein RA is hydrogen, C1-10 alkyl, C1-10 alkynyl, or C1-10 heteroalkyl, wherein the alkyl, alkynyl, or heteroalkyl is substituted with an oxo group (=O). In some embodiments, each instance of Y is independently –ORA, wherein RA is hydrogen, linear C1-10 alkyl, linear C1-10 alkynyl, or linear C1-10 heteroalkyl, wherein the alkyl, alkynyl, or heteroalkyl is substituted with an oxo group (=O) and/or an amino group (–NH2). In some embodiments, each instance of Y is independently – ORA, wherein RA is hydrogen, linear C1-10 alkyl, linear C1-10 alkynyl, or linear C1-10 heteroalkyl, wherein the alkyl, alkynyl, or heteroalkyl is substituted with an oxo group (=O). In some embodiments, each instance of Y is independently –ORA, wherein RA is hydrogen, C1-6 alkyl, C1-6 alkynyl, or C1-6 heteroalkyl, wherein the alkyl, alkynyl, or heteroalkyl is substituted with an oxo group (=O) and/or an amino group (–NH2). In some embodiments, each instance of Y is independently – ORA, wherein RA is hydrogen, C1-6 alkyl, C1-6 alkynyl, or C1-6 heteroalkyl, wherein the alkyl, alkynyl, or heteroalkyl is substituted with an oxo group (=O). In some embodiments, each instance of Y is independently –ORA, wherein RA is hydrogen, linear C1-6 alkyl, linear C1-6 alkynyl, or linear C1-6 heteroalkyl, wherein the alkyl, alkynyl, or heteroalkyl is substituted with an oxo group (=O) and/or an amino group (–NH2). In some embodiments, each instance of Y is independently –ORA, wherein RA is hydrogen, linear C1-6 alkyl, linear C1-6 alkynyl, or linear C1-6 heteroalkyl, wherein the alkyl, alkynyl, or heteroalkyl is substituted with an oxo group (=O).
U1197.70243WO00 89/193 #13587456v2 [0275] In some embodiments, at least one instance of Y is –OH, or [0276] In some embodiments, at least one instance of Y is –N(RA)2. In some embodiments, each instance of Y is independently –N(RA)2. In some embodiments, at least one instance of Y is –NHRA. In some embodiments, each instance of Y is independently –NHRA. In some embodiments, at least one instance of Y is –NH2. In some embodiments, each instance of Y is independently –NH2. [0277] In some embodiments, at least one instance of Y is –C(=O)ORA. In some embodiments, each instance of Y is independently –C(=O)ORA. In some embodiments, at least one instance of Y is – C(=O)OH. In some embodiments, each instance of Y is independently –C(=O)OH. [0278] As generally described herein, each instance of RA is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl. [0279] In some embodiments, at least one instance of RA is hydrogen, optionally substituted alkyl, optionally substituted alkynyl, or optionally substituted heteroalkyl. In some embodiments, at least one instance of RA is hydrogen or optionally substituted alkyl. In some embodiments, at least one instance of RA is hydrogen or optionally substituted alkynyl. In some embodiments, at least one instance of RA is hydrogen or optionally substituted heteroalkyl. In some embodiments, at least one instance of RA is hydrogen, optionally substituted alkyl, or optionally substituted alkynyl. In some embodiments, at least one instance of RA is hydrogen, optionally substituted alkyl, or optionally substituted heteroalkyl. In some embodiments, at least one instance of RA is hydrogen, optionally substituted alkynyl, or optionally substituted heteroalkyl. [0280] In some embodiments, each instance of RA is independently hydrogen, optionally substituted alkyl, optionally substituted alkynyl, or optionally substituted heteroalkyl. In some embodiments, each instance of RA is independently hydrogen or optionally substituted alkyl. In some embodiments, each instance of RA is independently hydrogen or optionally substituted alkynyl. In some embodiments, each instance of RA is independently hydrogen or optionally substituted heteroalkyl. In some
U1197.70243WO00 90/193 #13587456v2 embodiments, each instance of RA is independently hydrogen, optionally substituted alkyl, or optionally substituted alkynyl. In some embodiments, each instance of RA is independently hydrogen, optionally substituted alkyl, or optionally substituted heteroalkyl. In some embodiments, each instance of RA is independently hydrogen, optionally substituted alkynyl, or optionally substituted heteroalkyl. [0281] In some embodiments, at least one instance of RA is optionally substituted alkyl, optionally substituted alkynyl, or optionally substituted heteroalkyl. [0282] In some embodiments, at least one instance of RA is hydrogen, optionally substituted C1-10 alkyl, optionally substituted C1-10 alkynyl, or optionally substituted C1-10 heteroalkyl. In some embodiments, at least one instance of RA is hydrogen, optionally substituted linear C1-10 alkyl, optionally substituted linear C1-10 alkynyl, or optionally substituted linear C1-10 heteroalkyl. In some embodiments, at least one instance of RA is hydrogen, optionally substituted C1-6 alkyl, optionally substituted C1-6 alkynyl, or optionally substituted C1-6 heteroalkyl. In some embodiments, at least one instance of RA is hydrogen, optionally substituted linear C1-6 alkyl, optionally substituted linear C1-6 alkynyl, or optionally substituted linear C1-6 heteroalkyl. [0283] In some embodiments, at least one instance of RA is hydrogen, C1-10 alkyl, C1-10 alkynyl, or C1- 10 heteroalkyl, wherein the alkyl, alkynyl, or heteroalkyl is substituted with an oxo group (=O) and/or an amino group (–NH2). In some embodiments, at least one instance of RA is hydrogen, C1-10 alkyl, C1- 10 alkynyl, or C1-10 heteroalkyl, wherein the alkyl, alkynyl, or heteroalkyl is substituted with an oxo group (=O). In some embodiments, at least one instance of RA is hydrogen, linear C1-10 alkyl, linear C1-10 alkynyl, or linear C1-10 heteroalkyl, wherein the alkyl, alkynyl, or heteroalkyl is substituted with an oxo group (=O) and/or an amino group (–NH2). In some embodiments, at least one instance of RA is hydrogen, linear C1-10 alkyl, linear C1-10 alkynyl, or linear C1-10 heteroalkyl, wherein the alkyl, alkynyl, or heteroalkyl is substituted with an oxo group (=O). In some embodiments, at least one instance of RA is hydrogen, C1-6 alkyl, C1-6 alkynyl, or C1-6 heteroalkyl, wherein the alkyl, alkynyl, or heteroalkyl is substituted with an oxo group (=O) and/or an amino group (–NH2). In some embodiments, at least one instance of RA is hydrogen, C1-6 alkyl, C1-6 alkynyl, or C1-6 heteroalkyl, wherein the alkyl, alkynyl, or heteroalkyl is substituted with an oxo group (=O). In some embodiments, at least one instance of RA is hydrogen, linear C1-6 alkyl, linear C1-6 alkynyl, or linear C1-6 heteroalkyl, wherein the alkyl, alkynyl, or heteroalkyl is substituted with an oxo group (=O) and/or an amino group (–NH2). In some embodiments, at least one instance of RA is hydrogen, linear C1-6 alkyl, linear C1-6 alkynyl, or linear C1-6 heteroalkyl, wherein the alkyl, alkynyl, or heteroalkyl is substituted with an oxo group (=O). [0284] In some embodiments, each instance of RA is independently hydrogen, optionally substituted C1-10 alkyl, optionally substituted C1-10 alkynyl, or optionally substituted C1-10 heteroalkyl. In some embodiments, each instance of RA is independently hydrogen, optionally substituted linear C1-10 alkyl, optionally substituted linear C1-10 alkynyl, or optionally substituted linear C1-10 heteroalkyl. In some
U1197.70243WO00 91/193 #13587456v2 embodiments, each instance of RA is independently hydrogen, optionally substituted C1-6 alkyl, optionally substituted C1-6 alkynyl, or optionally substituted C1-6 heteroalkyl. In some embodiments, each instance of RA is independently hydrogen, optionally substituted linear C1-6 alkyl, optionally substituted linear C1-6 alkynyl, or optionally substituted linear C1-6 heteroalkyl. [0285] In some embodiments, each instance of RA is independently hydrogen, C1-10 alkyl, C1-10 alkynyl, or C1-10 heteroalkyl, wherein the alkyl, alkynyl, or heteroalkyl is substituted with an oxo group (=O) and/or an amino group (–NH2). In some embodiments, each instance of RA is independently hydrogen, C1-10 alkyl, C1-10 alkynyl, or C1-10 heteroalkyl, wherein the alkyl, alkynyl, or heteroalkyl is substituted with an oxo group (=O). In some embodiments, each instance of RA is independently hydrogen, linear C1-10 alkyl, linear C1-10 alkynyl, or linear C1-10 heteroalkyl, wherein the alkyl, alkynyl, or heteroalkyl is substituted with an oxo group (=O) and/or an amino group (–NH2). In some embodiments, each instance of RA is independently hydrogen, linear C1-10 alkyl, linear C1-10 alkynyl, or linear C1-10 heteroalkyl, wherein the alkyl, alkynyl, or heteroalkyl is substituted with an oxo group (=O). In some embodiments, each instance of RA is independently hydrogen, C1-6 alkyl, C1- 6 alkynyl, or C1-6 heteroalkyl, wherein the alkyl, alkynyl, or heteroalkyl is substituted with an oxo group (=O) and/or an amino group (–NH2). In some embodiments, each instance of RA is independently hydrogen, C1-6 alkyl, C1-6 alkynyl, or C1-6 heteroalkyl, wherein the alkyl, alkynyl, or heteroalkyl is substituted with an oxo group (=O). In some embodiments, each instance of RA is independently hydrogen, linear C1-6 alkyl, linear C1-6 alkynyl, or linear C1-6 heteroalkyl, wherein the alkyl, alkynyl, or heteroalkyl is substituted with an oxo group (=O) and/or an amino group (–NH2). In some embodiments, each instance of RA is independently hydrogen, linear C1-6 alkyl, linear C1-6 alkynyl, or linear C1-6 heteroalkyl, wherein the alkyl, alkynyl, or heteroalkyl is substituted with an oxo group (=O). [0286] In some embodiments, at least one instance of RA is hydrogen, or [0287] In some embodiments, at least one instance of RA is of formula: (i), wherein RA1 is optionally substituted alkyl, optionally substituted alkynyl, or optionally substituted heteroalkyl. In
U1197.70243WO00 92/193 #13587456v2 some embodiments, at least one instance of RA is of formula (i), wherein RA1 is optionally substituted C1-10 alkyl, optionally substituted C1-10 alkynyl, or optionally substituted C1-10 heteroalkyl. In some embodiments, at least one instance of RA is of formula (i), wherein RA1 is optionally substituted linear C1-10 alkyl, optionally substituted linear C1-10 alkynyl, or optionally substituted linear C1-10 heteroalkyl. In some embodiments, at least one instance of RA is of formula (i), wherein RA1 is optionally substituted C1-6 alkyl, optionally substituted C1-6 alkynyl, or optionally substituted C1-6 heteroalkyl. In some embodiments, at least one instance of RA is of formula (i), wherein RA1 is optionally substituted linear C1-6 alkyl, optionally substituted linear C1-6 alkynyl, or optionally substituted linear C1-6 heteroalkyl. In some embodiments, at least one instance of RA is of formula (i), wherein RA1 is [0288] In some embodiments, at least one instance of RA is hydrogen. In some embodiments, each instance of RA is hydrogen. [0289] In some embodiments, at least one instance of RA is optionally substituted alkyl. In some embodiments, at least one instance of RA is optionally substituted C1-10 alkyl. In some embodiments, at least one instance of RA is optionally substituted linear C1-10 alkyl. In some embodiments, at least one instance of RA is optionally substituted C1-6 alkyl. In some embodiments, at least one instance of RA is optionally substituted linear C1-6 alkyl. [0290] In some embodiments, at least one instance of RA is C1-10 alkyl substituted with an oxo group (=O) and/or an amino group (–NH2). In some embodiments, at least one instance of RA is C1-10 alkyl substituted with an oxo group (=O). In some embodiments, at least one instance of RA is linear C1-10 alkyl substituted with an oxo group (=O) and/or an amino group (–NH2). In some embodiments, at least one instance of RA is linear C1-10 alkyl substituted with an oxo group (=O). In some embodiments, at least one instance of RA is C1-6 alkyl substituted with an oxo group (=O) and/or an amino group (–NH2). In some embodiments, at least one instance of RA is C1-6 alkyl substituted with an oxo group (=O). In some embodiments, at least one instance of RA is linear C1-6 alkyl substituted with an oxo group (=O) and/or an amino group (–NH2). In some embodiments, at least one instance of RA is linear C1-6 alkyl substituted with an oxo group (=O). In some embodiments, at least one instance [0291] In some embodiments, each instance of RA is independently hydrogen or optionally substituted alkyl. In some embodiments, each instance of RA is independently hydrogen or optionally substituted C1-10 alkyl. In some embodiments, each instance of RA is independently hydrogen or optionally substituted linear C1-10 alkyl. In some embodiments, each instance of RA is independently hydrogen or optionally substituted C1-6 alkyl. In some embodiments, each instance of RA is independently hydrogen or optionally substituted linear C1-6 alkyl.
U1197.70243WO00 93/193 #13587456v2 [0292] In some embodiments, each instance of RA is independently hydrogen or C1-10 alkyl substituted with an oxo group (=O) and/or an amino group (–NH2). In some embodiments, each instance of RA is independently hydrogen or C1-10 alkyl substituted with an oxo group (=O). In some embodiments, each instance of RA is independently hydrogen or linear C1-10 alkyl substituted with an oxo group (=O) and/or an amino group (–NH2). In some embodiments, each instance of RA is independently hydrogen or linear C1-10 alkyl substituted with an oxo group (=O). In some embodiments, each instance of RA is independently hydrogen or C1-6 alkyl substituted with an oxo group (=O) and/or an amino group (–NH2). In some embodiments, each instance of RA is independently hydrogen or C1-6 alkyl substituted with an oxo group (=O). In some embodiments, each instance of RA is independently hydrogen or linear C1-6 alkyl substituted with an oxo group (=O) and/or an amino group (–NH2). In some embodiments, each instance of RA is independently hydrogen or linear C1-6 alkyl substituted with an oxo group (=O). In some embodiments, each instance of RA is independently hydrogen or . [0293] In some embodiments, at least one instance of RA is optionally substituted alkenyl. In some embodiments, at least one instance of RA is optionally substituted C1-10 alkenyl. In some embodiments, at least one instance of RA is optionally substituted linear C1-10 alkenyl. In some embodiments, at least one instance of RA is optionally substituted C1-6 alkenyl. In some embodiments, at least one instance of RA is optionally substituted linear C1-6 alkenyl. In some embodiments, each instance of RA is independently hydrogen or optionally substituted alkenyl. In some embodiments, each instance of RA is independently hydrogen or optionally substituted C1-10 alkenyl. In some embodiments, each instance of RA is independently hydrogen or optionally substituted linear C1-10 alkenyl. In some embodiments, each instance of RA is independently hydrogen or optionally substituted C1-6 alkenyl. In some embodiments, each instance of RA is independently hydrogen or optionally substituted linear C1-6 alkenyl. [0294] In some embodiments, at least one instance of RA is optionally substituted alkynyl. In some embodiments, at least one instance of RA is optionally substituted C1-10 alkynyl. In some embodiments, at least one instance of RA is optionally substituted linear C1-10 alkynyl. In some embodiments, at least one instance of RA is optionally substituted C1-6 alkynyl. In some embodiments, at least one instance of RA is optionally substituted linear C1-6 alkynyl. [0295] In some embodiments, at least one instance of RA is C1-10 alkynyl substituted with an oxo group (=O) and/or an amino group (–NH2). In some embodiments, at least one instance of RA is C1-10 alkynyl substituted with an oxo group (=O). In some embodiments, at least one instance of RA is linear C1-10 alkynyl substituted with an oxo group (=O) and/or an amino group (–NH2). In some embodiments, at least one instance of RA is linear C1-10 alkynyl substituted with an oxo group (=O). In some embodiments, at least one instance of RA is C1-6 alkynyl substituted with an oxo group (=O)
U1197.70243WO00 94/193 #13587456v2 and/or an amino group (–NH2). In some embodiments, at least one instance of RA is C1-6 alkynyl substituted with an oxo group (=O). In some embodiments, at least one instance of RA is linear C1-6 alkynyl substituted with an oxo group (=O) and/or an amino group (–NH2). In some embodiments, at least one instance of RA is linear C1-6 alkynyl substituted with an oxo group (=O). In some embodiments, at least one instance of RA is . [0296] In some embodiments, each instance of RA is independently hydrogen or optionally substituted C1-10 alkynyl. In some embodiments, each instance of RA is independently hydrogen or optionally substituted linear C1-10 alkynyl. In some embodiments, each instance of RA is independently hydrogen or optionally substituted C1-6 alkynyl. In some embodiments, each instance of RA is independently hydrogen or optionally substituted linear C1-6 alkynyl. [0297] In some embodiments, each instance of RA is independently hydrogen or C1-10 alkynyl substituted with an oxo group (=O) and/or an amino group (–NH2). In some embodiments, each instance of RA is independently hydrogen or C1-10 alkynyl substituted with an oxo group (=O). In some embodiments, each instance of RA is independently hydrogen or linear C1-10 alkynyl substituted with an oxo group (=O) and/or an amino group (–NH2). In some embodiments, each instance of RA is independently hydrogen or linear C1-10 alkynyl substituted with an oxo group (=O). In some embodiments, each instance of RA is independently hydrogen or C1-6 alkynyl substituted with an oxo group (=O) and/or an amino group (–NH2). In some embodiments, each instance of RA is independently hydrogen or C1-6 alkynyl substituted with an oxo group (=O). In some embodiments, each instance of RA is independently hydrogen or linear C1-6 alkynyl substituted with an oxo group (=O) and/or an amino group (–NH2). In some embodiments, each instance of RA is independently hydrogen or linear C1-6 alkynyl substituted with an oxo group (=O). In some embodiments, each instance of RA is independently hydrogen or [0298] In some embodiments, at least one instance of RA is optionally substituted heteroalkyl. In some embodiments, at least one instance of RA is optionally substituted C1-10 heteroalkyl. In some embodiments, at least one instance of RA is optionally substituted linear C1-10 heteroalkyl. In some embodiments, at least one instance of RA is optionally substituted C1-6 heteroalkyl. In some embodiments, at least one instance of RA is optionally substituted linear C1-6 heteroalkyl. [0299] In some embodiments, at least one instance of RA is C1-10 heteroalkyl substituted with an oxo group (=O) and/or an amino group (–NH2). In some embodiments, at least one instance of RA is C1-10 heteroalkyl substituted with an oxo group (=O). In some embodiments, at least one instance of RA is linear C1-10 heteroalkyl substituted with an oxo group (=O) and/or an amino group (–NH2). In some embodiments, at least one instance of RA is linear C1-10 heteroalkyl substituted with an oxo group
U1197.70243WO00 95/193 #13587456v2 (=O). In some embodiments, at least one instance of RA is C1-6 heteroalkyl substituted with an oxo group (=O) and/or an amino group (–NH2). In some embodiments, at least one instance of RA is C1-6 heteroalkyl substituted with an oxo group (=O). In some embodiments, at least one instance of RA is linear C1-6 heteroalkyl substituted with an oxo group (=O) and/or an amino group (–NH2). In some embodiments, at least one instance of RA is linear C1-6 heteroalkyl substituted with an oxo group (=O). In some embodiments, at least one instance of RA is . [0300] In some embodiments, each instance of RA is independently hydrogen or optionally substituted C1-10 heteroalkyl. In some embodiments, each instance of RA is independently hydrogen or optionally substituted linear C1-10 heteroalkyl. In some embodiments, each instance of RA is independently hydrogen or optionally substituted C1-6 heteroalkyl. In some embodiments, each instance of RA is independently hydrogen or optionally substituted linear C1-6 heteroalkyl. [0301] In some embodiments, each instance of RA is independently hydrogen or C1-10 heteroalkyl substituted with an oxo group (=O) and/or an amino group (–NH2). In some embodiments, each instance of RA is independently hydrogen or C1-10 heteroalkyl substituted with an oxo group (=O). In some embodiments, each instance of RA is independently hydrogen or linear C1-10 heteroalkyl substituted with an oxo group (=O) and/or an amino group (–NH2). In some embodiments, each instance of RA is independently hydrogen or linear C1-10 heteroalkyl substituted with an oxo group (=O). In some embodiments, each instance of RA is independently hydrogen or C1-6 heteroalkyl substituted with an oxo group (=O) and/or an amino group (–NH2). In some embodiments, each instance of RA is independently hydrogen or C1-6 heteroalkyl substituted with an oxo group (=O). In some embodiments, each instance of RA is independently hydrogen or linear C1-6 heteroalkyl substituted with an oxo group (=O) and/or an amino group (–NH2). In some embodiments, each instance of RA is independently hydrogen or linear C1-6 heteroalkyl substituted with an oxo group (=O). In some embodiments, each instance of RA is independently hydrogen or . [0302] In some embodiments, at least one instance of RA is optionally substituted heteroalkenyl. In some embodiments, at least one instance of RA is optionally substituted C1-10 heteroalkenyl. In some embodiments, at least one instance of RA is optionally substituted linear C1-10 heteroalkenyl. In some embodiments, at least one instance of RA is optionally substituted C1-6 heteroalkenyl. In some embodiments, at least one instance of RA is optionally substituted linear C1-6 heteroalkenyl. In some embodiments, each instance of RA is independently hydrogen or optionally substituted heteroalkenyl. In some embodiments, each instance of RA is independently hydrogen or optionally substituted C1-10 heteroalkenyl. In some embodiments, each instance of RA is independently hydrogen or optionally substituted linear C1-10 heteroalkenyl. In some embodiments, each instance of RA is independently
U1197.70243WO00 96/193 #13587456v2 hydrogen or optionally substituted C1-6 heteroalkenyl. In some embodiments, each instance of RA is independently hydrogen or optionally substituted linear C1-6 heteroalkenyl. [0303] In some embodiments, at least one instance of RA is optionally substituted heteroalkynyl. In some embodiments, at least one instance of RA is optionally substituted C1-10 heteroalkynyl. In some embodiments, at least one instance of RA is optionally substituted linear C1-10 heteroalkynyl. In some embodiments, at least one instance of RA is optionally substituted C1-6 heteroalkynyl. In some embodiments, at least one instance of RA is optionally substituted linear C1-6 heteroalkynyl. In some embodiments, each instance of RA is independently hydrogen or optionally substituted heteroalkynyl. In some embodiments, each instance of RA is independently hydrogen or optionally substituted C1-10 heteroalkynyl. In some embodiments, each instance of RA is independently hydrogen or optionally substituted linear C1-10 heteroalkynyl. In some embodiments, each instance of RA is independently hydrogen or optionally substituted C1-6 heteroalkynyl. In some embodiments, each instance of RA is independently hydrogen or optionally substituted linear C1-6 heteroalkynyl. [0304] In some embodiments, at least one instance of RA is optionally substituted carbocyclyl. In some embodiments, at least one instance of RA is optionally substituted C3-10 carbocyclyl. In some embodiments, at least one instance of RA is optionally substituted C4-6 carbocyclyl. In some embodiments, each instance of RA is independently hydrogen or optionally substituted carbocyclyl. In some embodiments, each instance of RA is independently hydrogen or optionally substituted C3-10 carbocyclyl. In some embodiments, each instance of RA is independently hydrogen or optionally substituted C4-6 carbocyclyl. [0305] In some embodiments, at least one instance of RA is optionally substituted heterocyclyl. In some embodiments, at least one instance of RA is optionally substituted 3-10 membered heterocyclyl. In some embodiments, at least one instance of RA is optionally substituted 4-6 membered heterocyclyl. In some embodiments, each instance of RA is independently hydrogen or optionally substituted heterocyclyl. In some embodiments, each instance of RA is independently hydrogen or optionally substituted 3-10 membered heterocyclyl. In some embodiments, each instance of RA is independently hydrogen or optionally substituted 4-6 membered heterocyclyl. [0306] In some embodiments, at least one instance of RA is optionally substituted aryl. In some embodiments, at least one instance of RA is optionally substituted phenyl. In some embodiments, each instance of RA is independently hydrogen or optionally substituted aryl. In some embodiments, each instance of RA is independently hydrogen or optionally substituted phenyl. [0307] In some embodiments, at least one instance of RA is optionally substituted heteroaryl. In some embodiments, at least one instance of RA is optionally substituted 5-10 membered heteroaryl. In some embodiments, at least one instance of RA is optionally substituted 5-6 membered heteroaryl. In some embodiments, at least one instance of RA is optionally substituted 5-6 monocyclic membered heteroaryl. In some embodiments, each instance of RA is independently hydrogen or optionally substituted heteroaryl. In some embodiments, each instance of RA is independently hydrogen or
U1197.70243WO00 97/193 #13587456v2 optionally substituted 5-10 membered heteroaryl. In some embodiments, each instance of RA is independently hydrogen or optionally substituted 5-6 membered heteroaryl. In some embodiments, each instance of RA is independently hydrogen or optionally substituted 5-6 monocyclic membered heteroaryl. [0308] In some embodiments, the compound of Formula (I) comprises an effective number of instances of Y for targeting a specific cell type. In some embodiments, the compound of Formula (I) comprises an effective number of instances of Y for targeting myeloid-derived suppressor cells (MDSCs). In some embodiments, the compound of Formula (I′) comprises an effective number of instances of Y for targeting a specific cell type. In some embodiments, the compound of Formula (I′) comprises an effective number of instances of Y for targeting myeloid-derived suppressor cells (MDSCs). Subgeneric Embodiments [0309] In some embodiments, the compound of Formula (I) or Formula (I′) is of Formula (I-a): or a pharmaceutically acceptable salt thereof. In some embodiments, the compound of Formula (I) is of Formula (I-a), or a pharmaceutically acceptable salt thereof. In some embodiments, the compound of Formula (I′) is of Formula (I-a), or a pharmaceutically acceptable salt thereof. [0310] In some embodiments, the compound of Formula (II) is of Formula (II-a): or a pharmaceutically acceptable salt thereof. [0311] In some embodiments, the compound of Formula (II) is of Formula (II-b): or a pharmaceutically acceptable salt thereof. [0312] In some embodiments, the compound of Formula (II) is of Formula (II-c):
U1197.70243WO00 98/193 #13587456v2 or a pharmaceutically acceptable salt thereof. [0313] In some embodiments, the compound of Formula (II) is of Formula (II-d) or Formula (II-e): or a pharmaceutically acceptable salt thereof. [0314] In some embodiments, the compound of Formula (II) is of Formula (II-d-i) or Formula (II-e-
U1197.70243WO00 99/193 #13587456v2 or a pharmaceutically acceptable salt thereof. [0315] In some embodiments, the compound of Formula (II) is of formula: , or a pharmaceutically acceptable salt thereof. [0316] In some embodiments, the compound of Formula (III) is of Formula (III-a): or a pharmaceutically acceptable salt thereof. [0317] In some embodiments, the compound of Formula (III) is of Formula (III-b):
U1197.70243WO00 100/193 #13587456v2 or a pharmaceutically acceptable salt thereof. [0318] In some embodiments, the compound of Formula (III) is of Formula (III-c): or a pharmaceutically acceptable salt thereof. [0319] In some embodiments, the compound of Formula (IV) is of Formula (IV-a): or a pharmaceutically acceptable salt thereof. [0320] In some embodiments, the compound of Formula (IV) is of Formula (IV-b): or a pharmaceutically acceptable salt thereof. [0321] In some embodiments, the compound of Formula (IV) is of Formula (IV-c): or a pharmaceutically acceptable salt thereof. [0322] In some embodiments, the compound of Formula (V) is of Formula (V-a): or a pharmaceutically acceptable salt thereof.
U1197.70243WO00 101/193 #13587456v2 [0323] In some embodiments, the compound of Formula (V) is of Formula (V-b): or a pharmaceutically acceptable salt thereof. [0324] In some embodiments, the compound of Formula (V) is of Formula (V-c): or a pharmaceutically acceptable salt thereof. [0325] In some embodiments, the compound of Formula (VI) is of Formula (VI-a): or a pharmaceutically acceptable salt thereof. [0326] In some embodiments, the compound of Formula (VI) is of Formula (VI-b): or a pharmaceutically acceptable salt thereof. [0327] In some embodiments, the compound of Formula (VI) is of Formula (VI-c): or a pharmaceutically acceptable salt thereof. [0328] In some embodiments, the compound of Formula (VII) is of Formula (VII-a): or a pharmaceutically acceptable salt thereof. [0329] In some embodiments, the compound of Formula (VII) is of Formula (VII-b):
U1197.70243WO00 102/193 #13587456v2 or a pharmaceutically acceptable salt thereof. [0330] In some embodiments, the compound of Formula (VII) is of Formula (VII-c): or a pharmaceutically acceptable salt thereof. [0331] In some embodiments, the compound of Formula (VIII) is of Formula (VIII-a): or a pharmaceutically acceptable salt thereof. [0332] In some embodiments, the compound of Formula (VIII) is of Formula (VIII-b): or a pharmaceutically acceptable salt thereof. [0333] In some embodiments, the compound of Formula (VIII) is of Formula (VIII-c):
U1197.70243WO00 103/193 #13587456v2 or a pharmaceutically acceptable salt thereof. [0334] In some embodiments, the compound of Formula (IX) is of Formula (IX-a): or a pharmaceutically acceptable salt thereof. [0335] In some embodiments, the compound of Formula (IX) is of Formula (IX-b): or a pharmaceutically acceptable salt thereof. [0336] In some embodiments, the compound of Formula (IX) is of Formula (IX-c):
U1197.70243WO00 104/193 #13587456v2
or a pharmaceutically acceptable salt thereof. [0337] In some embodiments, the compound of Formula (X) is of Formula (X-a): or a pharmaceutically acceptable salt thereof. [0338] In some embodiments, the compound of Formula (X) is of Formula (X-b): or a pharmaceutically acceptable salt thereof. [0339] In some embodiments, the compound of Formula (X) is of Formula (X-c): or a pharmaceutically acceptable salt thereof. [0340] In some embodiments, the compound of Formula (XI) is of Formula (XI-a):
U1197.70243WO00 105/193 #13587456v2 or a pharmaceutically acceptable salt thereof. [0341] In some embodiments, the compound of Formula (XI) is of Formula (XI-b): or a pharmaceutically acceptable salt thereof. [0342] In some embodiments, the compound of Formula (XI) is of Formula (XI-c): or a pharmaceutically acceptable salt thereof. [0343] In some embodiments, the compound of Formula (XI) is of Formula (XI-d):
U1197.70243WO00 106/193 #13587456v2 or a pharmaceutically acceptable salt thereof. [0344] In some embodiments, the compound of Formula (XI) is of Formula (XI-e): or a pharmaceutically acceptable salt thereof. [0345] In some embodiments, the compound of Formula (XI) is of formula: , or a pharmaceutically acceptable salt thereof. [0346] In some embodiments, a provided compound (a compound described herein, a compound of the present disclosure) is a compound of any of the formulae disclosed herein (e.g., Formulae (I), (I′), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), (XI)), or a pharmaceutically acceptable salt thereof. In some embodiments, a provided compound is a compound of any of the Formulae disclosed herein (e.g., Formulae (I), (I′), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), (XI)), or a salt thereof. In some embodiments, a provided compound is a compound of any of the Formulae disclosed herein (e.g., Formulae (I), (I′), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), (XI)). Compositions and Kits [0347] In some embodiments, the provided compound is provided in an effective amount in the pharmaceutical composition. In some embodiments, the effective amount is an amount effective for modulating a target protein in a subject or a cell, tissue, or biological sample. [0348] In some embodiments, the composition is formulated as a particle. In some embodiments, the composition is formulated as a nanoparticle or microparticle. In some embodiments, the particle is a microparticle (i.e., particle having a characteristic dimension of less than about 1 millimeter and at
U1197.70243WO00 107/193 #13587456v2 least about 1 micrometer, where the characteristic dimension of the particle is the smallest cross- sectional dimension of the particle). In some embodiments, the particle is a nanoparticle (i.e., a particle having a characteristic dimension of less than about 1 micrometer and at least about 1 nanometer, where the characteristic dimension of the particle is the smallest cross-sectional dimension of the particle). [0349] The particles described herein may include additional materials such as polymers (e.g., synthetic polymers (e.g., PEG, PLGA) and natural polymers (e.g., phospholipids)). In some embodiments, the additional materials are approved by a regulatory agency, such as the U.S. FDA, for human and veterinary use. [0350] The particles may be prepared using any method known in the art, such as precipitation, milling, spray drying, single and double emulsion solvent evaporation, solvent extraction, phase separation, and simple and complex coacervation. The conditions used in preparing the particles may be altered to yield particles of a desired size or property (e.g., hydrophobicity, hydrophilicity, external morphology, “stickiness”, shape, polydispersity, etc.). The method of preparing the particle and the conditions (e.g., solvent, temperature, concentration, and air flow rate, etc.) used may also depend on the agent being complexed, encapsulated, or mixed, and/or the composition of the matrix. [0351] Methods developed for making particles for delivery of agents that are included in the particles are described in the literature. See, e.g., Doubrow, M., Ed., “Microcapsules and Nanoparticles in Medicine and Pharmacy,” CRC Press, Boca Raton, 1992; Mathiowitz and Langer, J. Controlled Release 5:13-22, 1987; Mathiowitz et al., Reactive Polymers 6:275-283, 1987; Mathiowitz et al., J. Appl. Polymer Sci.35:755-774, 1988, which are incorporated by reference herein. [0352] If the particles prepared by any of the above methods have a size range outside of the desired range, the particles can be sized, for example, using a sieve. The particles may also be coated. In some embodiments, the particles are coated with a targeting agent. In some embodiments, the particles are coated with a surface-altering agent. In some embodiments, the particles are coated to achieve desirable surface properties (e.g., a particular charge). [0353] Pharmaceutical compositions described herein can be prepared by any method known in the art of pharmaceutics. In general, such preparatory methods include bringing the compound described herein (i.e., the “active ingredient”) into association with a carrier or excipient, and/or one or more other accessory ingredients, and then, if necessary and/or desirable, shaping, and/or packaging the product into a desired single- or multi-dose unit. [0354] Pharmaceutical compositions can be prepared, packaged, and/or sold in bulk, as a single unit dose, and/or as a plurality of single unit doses. A “unit dose” is a discrete amount of the pharmaceutical composition comprising a predetermined amount of the active ingredient. The amount of the active ingredient is generally equal to the dosage of the active ingredient which would be administered to a subject and/or a convenient fraction of such a dosage, such as one-half or one-third of such a dosage.
U1197.70243WO00 108/193 #13587456v2 [0355] Relative amounts of the active ingredient, the pharmaceutically acceptable excipient, and/or any additional ingredients in a pharmaceutical composition described herein will vary, depending upon the identity, size, and/or condition of the subject treated and further depending upon the route by which the composition is to be administered. The composition may comprise between 0.1% and 100% (w/w) active ingredient. [0356] Pharmaceutically acceptable excipients used in the manufacture of provided pharmaceutical compositions include inert diluents or fillers, dispersing and/or granulating agents, surface active agents and/or emulsifiers, disintegrating agents, binding agents, preservatives, buffering agents, lubricating agents, and/or oils. Excipients such as cocoa butter and suppository waxes, coloring agents, coating agents, sweetening, flavoring, and perfuming agents may also be present in the composition. [0357] Exemplary diluents or fillers include calcium carbonate, sodium carbonate, calcium phosphate, dicalcium phosphate, calcium sulfate, calcium hydrogen phosphate, sodium phosphate lactose, sucrose, cellulose, microcrystalline cellulose, kaolin, mannitol, sorbitol, inositol, sodium chloride, starches (such as dry starch, cornstarch), sugars (such as powdered sugar), calcium trisulfate, carboxymethylcellulose calcium, dextrate, dextrin, dextrose, fructose, lactitol, lactose, magnesium carbonate, magnesium, maltitol, maltodextrin, maltose, sucrose, glucose, mannitol, silicic acid, xylitol, and mixtures thereof. [0358] Exemplary granulating and/or dispersing agents include potato starch, corn starch, tapioca starch, sodium starch glycolate, clays, alginic acid, guar gum, citrus pulp, agar, bentonite, cellulose, and wood products, natural sponge, cation-exchange resins, calcium carbonate, silicates, sodium carbonate, cross-linked poly(vinyl-pyrrolidone) (crospovidone), sodium carboxymethyl starch (sodium starch glycolate), carboxymethyl cellulose, cross-linked sodium carboxymethyl cellulose (croscarmellose), methylcellulose, pregelatinized starch (starch 1500), microcrystalline starch, water insoluble starch, calcium carboxymethyl cellulose, magnesium aluminum silicate (Veegum), sodium lauryl sulfate, quaternary ammonium compounds, and mixtures thereof. [0359] Exemplary surface active agents and/or emulsifiers include natural emulsifiers (e.g., acacia, agar, alginic acid, sodium alginate, tragacanth, chondrux, cholesterol, xanthan, pectin, gelatin, egg yolk, casein, wool fat, cholesterol, wax, and lecithin), colloidal clays (e.g., bentonite (aluminum silicate) and Veegum (magnesium aluminum silicate)), long chain amino acid derivatives, high molecular weight alcohols (e.g., stearyl alcohol, cetyl alcohol, oleyl alcohol, triacetin monostearate, ethylene glycol distearate, glyceryl monostearate, and propylene glycol monostearate, polyvinyl alcohol), carbomers (e.g., carboxy polymethylene, polyacrylic acid, acrylic acid polymer, and carboxyvinyl polymer), carrageenan, cellulosic derivatives (e.g., carboxymethylcellulose sodium, powdered cellulose, hydroxymethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose, methylcellulose), sorbitan fatty acid esters (e.g., polyoxyethylene sorbitan monolaurate (Tween® 20), polyoxyethylene sorbitan (Tween® 60), polyoxyethylene sorbitan
U1197.70243WO00 109/193 #13587456v2 monooleate (Tween® 80), sorbitan monopalmitate (Span® 40), sorbitan monostearate (Span® 60), sorbitan tristearate (Span® 65), glyceryl monooleate, sorbitan monooleate (Span® 80), polyoxyethylene esters (e.g., polyoxyethylene monostearate (Myrj® 45), polyoxyethylene hydrogenated castor oil, polyethoxylated castor oil, polyoxymethylene stearate, and Solutol®), sucrose fatty acid esters, polyethylene glycol fatty acid esters (e.g., Cremophor®), polyoxyethylene ethers, (e.g., polyoxyethylene lauryl ether (Brij® 30)), poly(vinyl-pyrrolidone), diethylene glycol monolaurate, triethanolamine oleate, sodium oleate, potassium oleate, ethyl oleate, oleic acid, ethyl laurate, sodium lauryl sulfate, Pluronic® F-68, poloxamer P-188, cetrimonium bromide, cetylpyridinium chloride, benzalkonium chloride, docusate sodium, and/or mixtures thereof. [0360] Exemplary disintegrating agents or disintegrants include agar, algin, alginic acid, sodium alginate, silicates, sodium carbonate, calcium carbonate, carboxymethylcellulose, cellulose, clay, colloidal silicon dioxide, croscarmellose sodium, crospovidone, rubber, magnesium silicate, methylcellulose, potassium krillin, hydroxypropylcellulose (e.g., low substituted Hydroxypropylcellulose), crosslinked polyvinylpyrrolidone, hydroxypropylcellulose, and starch (e.g., sodium glycolate starch, potato or tapioca starch). [0361] Exemplary binding agents include starch (e.g., glycolate starch, cornstarch and starch paste), gelatin, sugars (e.g., sucrose, glucose, dextrose, dextrin, molasses, lactose, lactitol, mannitol, etc.), natural and synthetic gums (e.g., acacia, sodium alginate, extract of Irish moss, panwar gum, ghatti gum, mucilage of isapol husks, carboxymethylcellulose, methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose, microcrystalline cellulose, cellulose acetate, poly(vinyl-pyrrolidone), magnesium aluminum silicate (Veegum®), and larch arabogalactan), alginates, polyethylene oxide, polyethylene glycol, inorganic calcium salts, silicic acid, polymethacrylates, waxes, water, alcohol, and/or mixtures thereof. [0362] Exemplary preservatives include antioxidants, chelating agents, antimicrobial preservatives, antifungal preservatives, antiprotozoan preservatives, alcohol preservatives, acidic preservatives, and other preservatives. In certain embodiments, the preservative is an antioxidant. In other embodiments, the preservative is a chelating agent. [0363] Exemplary antioxidants include alpha tocopherol, ascorbic acid, ascorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene, monothioglycerol, potassium metabisulfite, propionic acid, propyl gallate, sodium ascorbate, sodium bisulfite, sodium metabisulfite, and sodium sulfite. [0364] Exemplary chelating agents include ethylenediaminetetraacetic acid (EDTA) and salts and hydrates thereof (e.g., sodium edetate, disodium edetate, trisodium edetate, calcium disodium edetate, dipotassium edetate, and the like), citric acid and salts and hydrates thereof (e.g., citric acid monohydrate), fumaric acid and salts and hydrates thereof, malic acid and salts and hydrates thereof, phosphoric acid and salts and hydrates thereof, and tartaric acid and salts and hydrates thereof. [0365] Exemplary antimicrobial preservatives include benzalkonium chloride, benzethonium chloride, benzyl alcohol, bronopol, cetrimide, cetylpyridinium chloride, chlorhexidine, chlorobutanol,
U1197.70243WO00 110/193 #13587456v2 chlorocresol, chloroxylenol, cresol, ethyl alcohol, glycerin, hexetidine, imidurea, phenol, phenoxyethanol, phenylethyl alcohol, phenylmercuric nitrate, propylene glycol, and thimerosal. [0366] Exemplary antifungal preservatives include butyl paraben, methyl paraben, ethyl paraben, propyl paraben, benzoic acid, hydroxybenzoic acid, potassium benzoate, potassium sorbate, sodium benzoate, sodium propionate, and sorbic acid. [0367] Exemplary alcohol preservatives include ethanol, polyethylene glycol, phenol, phenolic compounds, bisphenol, chlorobutanol, hydroxybenzoate, and phenylethyl alcohol. [0368] Exemplary acidic preservatives include vitamin A, vitamin C, vitamin E, beta-carotene, citric acid, acetic acid, dehydroacetic acid, ascorbic acid, sorbic acid, and phytic acid. [0369] Other preservatives include tocopherol, tocopherol acetate, deteroxime mesylate, cetrimide, butylated hydroxyanisol (BHA), butylated hydroxytoluened (BHT), ethylenediamine, sodium lauryl sulfate (SLS), sodium lauryl ether sulfate (SLES), sodium bisulfite, sodium metabisulfite, potassium sulfite, potassium metabisulfite, Glydant® Plus, Phenonip®, methylparaben, Germall® 115, Germaben® II, Neolone®, Kathon®, and Euxyl®. [0370] Exemplary buffering agents include citrate buffer solutions, acetate buffer solutions, phosphate buffer solutions, ammonium chloride, calcium carbonate, calcium chloride, calcium citrate, calcium glubionate, calcium gluceptate, calcium gluconate, D-gluconic acid, calcium glycerophosphate, calcium lactate, propanoic acid, calcium levulinate, pentanoic acid, dibasic calcium phosphate, phosphoric acid, tribasic calcium phosphate, calcium hydroxide phosphate, potassium acetate, potassium chloride, potassium gluconate, potassium mixtures, dibasic potassium phosphate, monobasic potassium phosphate, potassium phosphate mixtures, sodium acetate, sodium bicarbonate, sodium chloride, sodium citrate, sodium lactate, dibasic sodium phosphate, monobasic sodium phosphate, sodium phosphate mixtures, tromethamine, magnesium hydroxide, aluminum hydroxide, alginic acid, pyrogen-free water, isotonic saline, Ringer’s solution, ethyl alcohol, and mixtures thereof. [0371] Exemplary lubricating agents include agar, ethyl oleate, ethyl laurate, glycerin, blyceryl palmitostearate, magnesium oxide, magnesium stearate, mannitol, poloxamer, glycol, sodium stearyl, sorbitol, zinc stearate, magnesium stearate, calcium stearate, stearic acid, silica, talc, malt, glyceryl behanate, hydrogenated vegetable oils, polyethylene glycol, sodium benzoate, sodium acetate, sodium chloride, leucine, magnesium lauryl sulfate, sodium lauryl sulfate, and mixtures thereof. [0372] Exemplary natural oils include almond, apricot kernel, avocado, babassu, bergamot, black current seed, borage, cade, camomile, canola, caraway, carnauba, castor, cinnamon, cocoa butter, coconut, cod liver, coffee, corn, cotton seed, emu, eucalyptus, evening primrose, fish, flaxseed, geraniol, gourd, grape seed, hazel nut, hyssop, isopropyl myristate, jojoba, kukui nut, lavandin, lavender, lemon, litsea cubeba, macademia nut, mallow, mango seed, meadowfoam seed, mink, nutmeg, olive, orange, orange roughy, palm, palm kernel, peach kernel, peanut, poppy seed, pumpkin seed, rapeseed, rice bran, rosemary, safflower, sandalwood, sasquana, savoury, sea buckthorn,
U1197.70243WO00 111/193 #13587456v2 sesame, shea butter, silicone, soybean, sunflower, tea tree, thistle, tsubaki, vetiver, walnut, and wheat germ oils. Exemplary synthetic oils include, but are not limited to, butyl stearate, caprylic triglyceride, capric triglyceride, cyclomethicone, diethyl sebacate, dimethicone 360, isopropyl myristate, mineral oil, octyldodecanol, oleyl alcohol, silicone oil, and mixtures thereof. [0373] Liquid dosage forms for oral and parenteral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active ingredients, the liquid dosage forms may comprise inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (e.g., cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof. Besides inert diluents, the oral compositions can include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents. In certain embodiments for parenteral administration, the conjugates described herein are mixed with solubilizing agents such as Cremophor®, alcohols, oils, modified oils, glycols, polysorbates, cyclodextrins, polymers, and mixtures thereof. [0374] Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions can be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation can be a sterile injectable solution, suspension, or emulsion in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that can be employed are water, Ringer’s solution, U.S.P., and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil can be employed including synthetic mono- or di-glycerides. In addition, fatty acids such as oleic acid are used in the preparation of injectables. [0375] In some embodiments, injectable preparations of the compositions disclosed herein are in the form of a ready-to-use (“RTU”) preparation that can be directly administered to a subject. In some embodiments, the RTU preparation is a suspension. In some embodiments, the RTU preparation is a solution. In some embodiments, the RTU preparation is an emulsion. In some embodiments, injectable preparations of the compositions disclosed herein are in the form of a solid that is reconstituted prior to administration. In some embodiments, the solid is a lyophilized solid. In some embodiments, injectable preparations of the compositions disclosed herein are in the form of a liquid or suspension that is diluted prior to administration. [0376] The injectable formulations can be sterilized, for example, by filtration through a bacterial- retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.
U1197.70243WO00 112/193 #13587456v2 [0377] In order to prolong the effect of a drug, it is often desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. This can be accomplished by the use of a liquid suspension of crystalline or amorphous material with poor water solubility. The rate of absorption of the drug then depends upon its rate of dissolution, which, in turn, may depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered drug form may be accomplished by dissolving or suspending the drug in an oil vehicle. [0378] Compositions for rectal or vaginal administration are typically suppositories which can be prepared by mixing the conjugates described herein with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol, or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active ingredient. [0379] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active ingredient is mixed with at least one inert, pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate and/or (a) fillers or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic acid, (b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone, sucrose, and acacia, (c) humectants such as glycerol, (d) disintegrating agents such as agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, (e) solution retarding agents such as paraffin, (f) absorption accelerators such as quaternary ammonium compounds, (g) wetting agents such as, for example, cetyl alcohol and glycerol monostearate, (h) absorbents such as kaolin and bentonite clay, and (i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets, and pills, the dosage form may include a buffering agent. [0380] Solid compositions of a similar type can be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the art of pharmacology. They may optionally comprise opacifying agents and can be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of encapsulating compositions which can be used include polymeric substances and waxes. Solid compositions of a similar type can be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polethylene glycols and the like. [0381] The active ingredient can be in a micro-encapsulated form with one or more excipients as noted above. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings, release controlling coatings, and other coatings well known in the pharmaceutical formulating art. In such solid dosage forms the active ingredient can be
U1197.70243WO00 113/193 #13587456v2 admixed with at least one inert diluent such as sucrose, lactose, or starch. Such dosage forms may comprise, as is normal practice, additional substances other than inert diluents, e.g., tableting lubricants and other tableting aids such a magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets and pills, the dosage forms may comprise buffering agents. They may optionally comprise opacifying agents and can be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of encapsulating agents which can be used include polymeric substances and waxes. [0382] Dosage forms for topical and/or transdermal administration of a compound described herein may include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants, and/or patches. Generally, the active ingredient is admixed under sterile conditions with a pharmaceutically acceptable carrier or excipient and/or any needed preservatives and/or buffers as can be required. Additionally, the present disclosure contemplates the use of transdermal patches, which often have the added advantage of providing controlled delivery of an active ingredient to the body. Such dosage forms can be prepared, for example, by dissolving and/or dispensing the active ingredient in the proper medium. Alternatively or additionally, the rate can be controlled by either providing a rate controlling membrane and/or by dispersing the active ingredient in a polymer matrix and/or gel. [0383] Suitable devices for use in delivering intradermal pharmaceutical compositions described herein include short needle devices. Intradermal compositions can be administered by devices which limit the effective penetration length of a needle into the skin. Alternatively or additionally, conventional syringes can be used in the classical mantoux method of intradermal administration. Jet injection devices which deliver liquid formulations to the dermis via a liquid jet injector and/or via a needle which pierces the stratum corneum and produces a jet which reaches the dermis are suitable. Ballistic powder/particle delivery devices which use compressed gas to accelerate the compound in powder form through the outer layers of the skin to the dermis are suitable. [0384] Formulations suitable for topical administration include, but are not limited to, liquid and/or semi-liquid preparations such as liniments, lotions, oil-in-water and/or water-in-oil emulsions such as creams, ointments, and/or pastes, and/or solutions and/or suspensions. Topically administrable formulations may, for example, comprise from about 1% to about 10% (w/w) active ingredient, although the concentration of the active ingredient can be as high as the solubility limit of the active ingredient in the solvent. Formulations for topical administration may further comprise one or more of the additional ingredients described herein. [0385] A pharmaceutical composition described herein can be prepared, packaged, and/or sold in a formulation suitable for pulmonary administration via the buccal cavity. Such a formulation may comprise dry particles which comprise the active ingredient and which have a diameter in the range from about 0.5 to about 7 nanometers, or from about 1 to about 6 nanometers. Such compositions are conveniently in the form of dry powders for administration using a device comprising a dry powder
U1197.70243WO00 114/193 #13587456v2 reservoir to which a stream of propellant can be directed to disperse the powder and/or using a self- propelling solvent/powder dispensing container such as a device comprising the active ingredient dissolved and/or suspended in a low-boiling propellant in a sealed container. Such powders comprise particles wherein at least 98% of the particles by weight have a diameter greater than 0.5 nanometers and at least 95% of the particles by number have a diameter less than 7 nanometers. Alternatively, at least 95% of the particles by weight have a diameter greater than 1 nanometer and at least 90% of the particles by number have a diameter less than 6 nanometers. Dry powder compositions may include a solid fine powder diluent such as sugar and are conveniently provided in a unit dose form. [0386] Low boiling propellants generally include liquid propellants having a boiling point of below 65 °F at atmospheric pressure. Generally the propellant may constitute 50 to 99.9% (w/w) of the composition, and the active ingredient may constitute 0.1 to 20% (w/w) of the composition. The propellant may further comprise additional ingredients such as a liquid non-ionic and/or solid anionic surfactant and/or a solid diluent (which may have a particle size of the same order as particles comprising the active ingredient). [0387] Pharmaceutical compositions described herein formulated for pulmonary delivery may provide the active ingredient in the form of droplets of a solution and/or suspension. Such formulations can be prepared, packaged, and/or sold as aqueous and/or dilute alcoholic solutions and/or suspensions, optionally sterile, comprising the active ingredient, and may conveniently be administered using any nebulization and/or atomization device. Such formulations may further comprise one or more additional ingredients including, but not limited to, a flavoring agent such as saccharin sodium, a volatile oil, a buffering agent, a surface active agent, and/or a preservative such as methylhydroxybenzoate. The droplets provided by this route of administration may have an average diameter in the range from about 0.1 to about 200 nanometers. [0388] Formulations described herein as being useful for pulmonary delivery are useful for intranasal delivery of a pharmaceutical composition described herein. Another formulation suitable for intranasal administration is a coarse powder comprising the active ingredient and having an average particle from about 0.2 to 500 micrometers. Such a formulation is administered by rapid inhalation through the nasal passage from a container of the powder held close to the nares. [0389] Formulations for nasal administration may, for example, comprise from about as little as 0.1% (w/w) to as much as 100% (w/w) of the active ingredient, and may comprise one or more of the additional ingredients described herein. A pharmaceutical composition described herein can be prepared, packaged, and/or sold in a formulation for buccal administration. Such formulations may, for example, be in the form of tablets and/or lozenges made using conventional methods, and may contain, for example, 0.1 to 20% (w/w) active ingredient, the balance comprising an orally dissolvable and/or degradable composition and, optionally, one or more of the additional ingredients described herein. Alternately, formulations for buccal administration may comprise a powder and/or an aerosolized and/or atomized solution and/or suspension comprising the active ingredient. Such
U1197.70243WO00 115/193 #13587456v2 powdered, aerosolized, and/or aerosolized formulations, when dispersed, may have an average particle and/or droplet size in the range from about 0.1 to about 200 nanometers, and may further comprise one or more of the additional ingredients described herein. [0390] A pharmaceutical composition described herein can be prepared, packaged, and/or sold in a formulation for ophthalmic administration. Such formulations may, for example, be in the form of eye drops including, for example, a 0.1-1.0% (w/w) solution and/or suspension of the active ingredient in an aqueous or oily liquid carrier or excipient. Such drops may further comprise buffering agents, salts, and/or one or more other of the additional ingredients described herein. Other opthalmically- administrable formulations which are useful include those which comprise the active ingredient in microcrystalline form and/or in a liposomal preparation. Ear drops and/or eye drops are also contemplated as being within the scope of this disclosure. [0391] Although the descriptions of pharmaceutical compositions provided herein are principally directed to pharmaceutical compositions which are suitable for administration to humans, it will be understood by the skilled artisan that such compositions are generally suitable for administration to animals of all sorts. Modification of pharmaceutical compositions suitable for administration to humans in order to render the compositions suitable for administration to various animals is well understood, and the ordinarily skilled veterinary pharmacologist can design and/or perform such modification with ordinary experimentation. [0392] Compounds provided herein are typically formulated in dosage unit form for ease of administration and uniformity of dosage. It will be understood, however, that the total daily usage of the compositions described herein will be decided by a physician within the scope of sound medical judgment. The specific therapeutically effective dose level for any particular subject or organism will depend upon a variety of factors including the disease being treated and the severity of the disorder; the activity of the specific active ingredient employed; the specific composition employed; the age, body weight, general health, sex, and diet of the subject; the time of administration, route of administration, and rate of excretion of the specific active ingredient employed; the duration of the treatment; drugs used in combination or coincidental with the specific active ingredient employed; and like factors well known in the medical arts. [0393] The compounds and compositions provided herein can be administered by any route, including enteral (e.g., oral), parenteral, intravenous, intramuscular, intra-arterial, intramedullary, intrathecal, subcutaneous, intraventricular, transdermal, interdermal, rectal, intravaginal, intraperitoneal, topical (as by powders, ointments, creams, and/or drops), mucosal, nasal, bucal, sublingual; by intratracheal instillation, bronchial instillation, and/or inhalation; and/or as an oral spray, nasal spray, and/or aerosol. Specifically contemplated routes are oral administration, intravenous administration (e.g., systemic intravenous injection), regional administration via blood and/or lymph supply, and/or direct administration to an affected site. In general, the most appropriate route of administration will depend upon a variety of factors including the nature of the agent (e.g., its
U1197.70243WO00 116/193 #13587456v2 stability in the environment of the gastrointestinal tract), and/or the condition of the subject (e.g., whether the subject is able to tolerate oral administration). In certain embodiments, the compound or pharmaceutical composition described herein is suitable for topical administration to the eye of a subject. [0394] The exact amount of a compound required to achieve an effective amount will vary from subject to subject, depending, for example, on species, age, and general condition of a subject, severity of the side effects or disorder, identity of the particular compound, mode of administration, and the like. An effective amount may be included in a single dose (e.g., single oral dose) or multiple doses (e.g., multiple oral doses). In certain embodiments, when multiple doses are administered to a subject or applied to a tissue or cell, any two doses of the multiple doses include different or substantially the same amounts of a compound described herein. In certain embodiments, when multiple doses are administered to a subject or applied to a tissue or cell, the frequency of administering the multiple doses to the subject or applying the multiple doses to the tissue or cell is three doses a day, two doses a day, one dose a day, one dose every other day, one dose every third day, one dose every week, one dose every two weeks, one dose every three weeks, or one dose every four weeks. In certain embodiments, the frequency of administering the multiple doses to the subject or applying the multiple doses to the tissue or cell is one dose per day. In certain embodiments, the frequency of administering the multiple doses to the subject or applying the multiple doses to the tissue or cell is two doses per day. In certain embodiments, the frequency of administering the multiple doses to the subject or applying the multiple doses to the tissue or cell is three doses per day. In certain embodiments, when multiple doses are administered to a subject or applied to a tissue or cell, the duration between the first dose and last dose of the multiple doses is one day, two days, four days, one week, two weeks, three weeks, one month, two months, three months, four months, six months, nine months, one year, two years, three years, four years, five years, seven years, ten years, fifteen years, twenty years, or the lifetime of the subject, tissue, or cell. In certain embodiments, the duration between the first dose and last dose of the multiple doses is three months, six months, or one year. In certain embodiments, the duration between the first dose and last dose of the multiple doses is the lifetime of the subject, tissue, or cell. In certain embodiments, a dose (e.g., a single dose, or any dose of multiple doses) described herein includes independently between 0.1 µg and 1 µg, between 0.001 mg and 0.01 mg, between 0.01 mg and 0.1 mg, between 0.1 mg and 1 mg, between 1 mg and 3 mg, between 3 mg and 10 mg, between 10 mg and 30 mg, between 30 mg and 100 mg, between 100 mg and 300 mg, between 300 mg and 1,000 mg, or between 1 g and 10 g, inclusive, of a compound described herein. In certain embodiments, a dose described herein includes independently between 1 mg and 3 mg, inclusive, of a compound described herein. In certain embodiments, a dose described herein includes independently between 3 mg and 10 mg, inclusive, of a compound described herein. In certain embodiments, a dose described herein includes independently between 10
U1197.70243WO00 117/193 #13587456v2 mg and 30 mg, inclusive, of a compound described herein. In certain embodiments, a dose described herein includes independently between 30 mg and 100 mg, inclusive, of a compound described herein. [0395] Dose ranges as described herein provide guidance for the administration of provided pharmaceutical compositions to an adult. The amount to be administered to, for example, a child or an adolescent can be determined by a medical practitioner or person skilled in the art and can be lower or the same as that administered to an adult. [0396] A compound or composition, as described herein, can be administered in combination with one or more additional pharmaceutical agents (e.g., therapeutically and/or prophylactically active agents). The compounds or compositions can be administered in combination with additional pharmaceutical agents that improve their activity (e.g., activity (e.g., potency and/or efficacy) in treating a disease or disorder in a subject in need thereof, in preventing a disease or disorder in a subject in need thereof, in reducing the risk to develop a disease or disorder in a subject in need thereof, and/or in cellular engineering in a subject or cell), improve bioavailability, improve safety, reduce drug resistance, reduce and/or modify metabolism, inhibit excretion, and/or modify distribution in a subject or cell. It will also be appreciated that the therapy employed may achieve a desired effect for the same disorder, and/or it may achieve different effects. In certain embodiments, a pharmaceutical composition described herein including a compound described herein and an additional pharmaceutical agent shows a synergistic effect that is absent in a pharmaceutical composition including one of the compound and the additional pharmaceutical agent, but not both. In some embodiments, the additional pharmaceutical agent achieves a desired effect for the same disorder. In some embodiments, the additional pharmaceutical agent achieves different effects. [0397] The compound or composition can be administered concurrently with, prior to, or subsequent to one or more additional pharmaceutical agents, which may be useful as, e.g., combination therapies. Pharmaceutical agents include therapeutically active agents. Pharmaceutical agents also include prophylactically active agents. Pharmaceutical agents include small organic molecules such as drug compounds (e.g., compounds approved for human or veterinary use by the U.S. Food and Drug Administration as provided in the Code of Federal Regulations (CFR)), peptides, proteins, carbohydrates, monosaccharides, oligosaccharides, polysaccharides, nucleoproteins, mucoproteins, lipoproteins, synthetic polypeptides or proteins, small molecules linked to proteins, glycoproteins, steroids, nucleic acids, DNAs, RNAs, nucleotides, nucleosides, oligonucleotides, antisense oligonucleotides, lipids, hormones, vitamins, and cells. In certain embodiments, the additional pharmaceutical agent is a pharmaceutical agent useful for treating and/or preventing a disease or disorder (e.g., proliferative disease, hematological disease, neurological disease, painful condition, psychiatric disorder, or metabolic disorder). Each additional pharmaceutical agent may be administered at a dose and/or on a time schedule determined for that pharmaceutical agent. The additional pharmaceutical agents may also be administered together with each other and/or with the compound or composition described herein in a single dose or composition or administered separately
U1197.70243WO00 118/193 #13587456v2 in different doses or compositions. The particular combination to employ in a regimen will take into account compatibility of the compound described herein with the additional pharmaceutical agent(s) and/or the desired therapeutic and/or prophylactic effect to be achieved. In general, it is expected that the additional pharmaceutical agent(s) in combination be utilized at levels that do not exceed the levels at which they are utilized individually. In some embodiments, the levels utilized in combination will be lower than those utilized individually. [0398] The additional pharmaceutical agents include, but are not limited to, anti-proliferative agents, anti-cancer agents, anti-angiogenesis agents, steroidal or non-steroidal anti-inflammatory agents, immunosuppressants, anti-bacterial agents, anti-viral agents, cardiovascular agents, cholesterol- lowering agents, anti-diabetic agents, anti-allergic agents, contraceptive agents, pain-relieving agents, anesthetics, anti–coagulants, inhibitors of an enzyme, steroidal agents, steroidal or antihistamine, antigens, vaccines, antibodies, decongestant, sedatives, opioids, analgesics, anti–pyretics, hormones, and prostaglandins. In certain embodiments, the additional pharmaceutical agent is an anti- proliferative agent. In certain embodiments, the additional pharmaceutical agent is an anti-cancer agent. In certain embodiments, the additional pharmaceutical agent is an anti-viral agent. In certain embodiments, the additional pharmaceutical agent is an binder or inhibitor of a protein kinase. In certain embodiments, the additional pharmaceutical agent is selected from the group consisting of epigenetic or transcriptional modulators (e.g., DNA methyltransferase inhibitors, histone deacetylase inhibitors (HDAC inhibitors), lysine methyltransferase inhibitors), antimitotic drugs (e.g., taxanes and vinca alkaloids), hormone receptor modulators (e.g., estrogen receptor modulators and androgen receptor modulators), cell signaling pathway inhibitors (e.g., tyrosine protein kinase inhibitors), modulators of protein stability (e.g., proteasome inhibitors), Hsp90 inhibitors, glucocorticoids, all- trans retinoic acids, and other agents that promote differentiation. In certain embodiments, the compounds described herein or pharmaceutical compositions can be administered in combination with an anti-cancer therapy including, but not limited to, surgery, radiation therapy, transplantation (e.g., stem cell transplantation, bone marrow transplantation), immunotherapy, and chemotherapy. Additional pharmaceutical agents include small organic molecules such as drug compounds (e.g., compounds approved by the US Food and Drug Administration as provided in the Code of Federal Regulations (CFR)), peptides, proteins, carbohydrates, monosaccharides, oligosaccharides, polysaccharides, nucleoproteins, mucoproteins, lipoproteins, synthetic polypeptides or proteins, small molecules linked to proteins, glycoproteins, steroids, nucleic acids, DNAs, RNAs, nucleotides, nucleosides, oligonucleotides, antisense oligonucleotides, lipids, hormones, vitamins and cells. [0399] Also encompassed by the disclosure are kits (e.g., pharmaceutical packs). The kits provided may comprise a pharmaceutical composition or compound described herein and a container (e.g., a vial, ampule, bottle, syringe, and/or dispenser package, or other suitable container). In some embodiments, provided kits may optionally further include a second container comprising a pharmaceutical excipient for dilution or suspension of a pharmaceutical composition or compound
U1197.70243WO00 119/193 #13587456v2 described herein. In some embodiments, the pharmaceutical composition or compound described herein provided in the first container and the second container are combined to form one unit dosage form. [0400] Thus, in one aspect, provided are kits including a first container comprising a compound or pharmaceutical composition described herein. In certain embodiments, the kits are useful for treating a disease or disorder (e.g., proliferative disease, hematological disease, neurological disease, painful condition, psychiatric disorder, or metabolic disorder) in a subject in need thereof. In certain embodiments, the kits are useful for preventing a disease or disorder (e.g., proliferative disease, hematological disease, neurological disease, painful condition, psychiatric disorder, or metabolic disorder) in a subject in need thereof. In certain embodiments, the kits are useful for reducing the risk of developing a disease or disorder (e.g., proliferative disease, hematological disease, neurological disease, painful condition, psychiatric disorder, or metabolic disorder) in a subject in need thereof. In certain embodiments, the kits are useful for cellular engineering in a subject or cell. [0401] In certain embodiments, a kit described herein further includes instructions for using the kit. A kit described herein may also include information as required by a regulatory agency such as the U.S. Food and Drug Administration (FDA). In certain embodiments, the information included in the kits is prescribing information. In certain embodiments, the kits and instructions provide for treating a disease or disorder (e.g., proliferative disease, hematological disease, neurological disease, painful condition, psychiatric disorder, or metabolic disorder) in a subject in need thereof. In certain embodiments, the kits and instructions provide for preventing a disease or disorder (e.g., proliferative disease, hematological disease, neurological disease, painful condition, psychiatric disorder, or metabolic disorder) in a subject in need thereof. In certain embodiments, the kits and instructions provide for reducing the risk of developing a disease or disorder (e.g., proliferative disease, hematological disease, neurological disease, painful condition, psychiatric disorder, or metabolic disorder) in a subject in need thereof. In certain embodiments, the kits and instructions provide for cellular engineering in a subject or cell. A kit described herein may include one or more additional pharmaceutical agents described herein as a separate composition. [0402] In some embodiments, a provided composition (a composition described herein, a composition of the present disclosure) is a composition comprising a compound of any of the formulae herein (e.g., Formulae (I), (I′), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), (XI)), or a pharmaceutically acceptable salt thereof. In some embodiments, a provided composition is a composition comprising a compound of any of the formulae herein (e.g., Formulae (I), (I′), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), (XI)), or a salt thereof. In some embodiments, a provided composition is a composition comprising a compound of any of the formulae herein (e.g., Formulae (I), (I′), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), (XI)).
U1197.70243WO00 120/193 #13587456v2 Methods of Treatment and Prevention and Methods of Modulating a Target Protein [0403] In another aspect, the present disclosure provides a method of treating or preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a provided compound or provided composition. In some embodiments, the present disclosure provides a method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a provided compound or provided composition. In some embodiments, the present disclosure provides a method of preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a provided compound or provided composition. [0404] In another aspect, the present disclosure provides a method of treating or preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formulae (I), (I′), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), or (XI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the present disclosure provides a method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formulae (I), (I′), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), or (XI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the present disclosure provides a method of preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formulae (I), (I′), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), or (XI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. [0405] In some embodiments, the present disclosure provides a method of treating or preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the present disclosure provides a method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the present disclosure provides a method of preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. [0406] In some embodiments, the present disclosure provides a method of treating or preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (I′), or a pharmaceutically
U1197.70243WO00 121/193 #13587456v2 acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the present disclosure provides a method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (I′), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the present disclosure provides a method of preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (I′), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. [0407] In some embodiments, the present disclosure provides a method of treating or preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (II), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the present disclosure provides a method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (II), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the present disclosure provides a method of preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (II), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. [0408] In some embodiments, the present disclosure provides a method of treating or preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (III), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the present disclosure provides a method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (III), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the present disclosure provides a method of preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (III), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. [0409] In some embodiments, the present disclosure provides a method of treating or preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (IV), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the present disclosure provides a method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (IV), or a pharmaceutically acceptable salt thereof, or a pharmaceutical
U1197.70243WO00 122/193 #13587456v2 composition thereof. In some embodiments, the present disclosure provides a method of preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (IV), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. [0410] In some embodiments, the present disclosure provides a method of treating or preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (V), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the present disclosure provides a method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (V), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the present disclosure provides a method of preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (V), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. [0411] In some embodiments, the present disclosure provides a method of treating or preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (VI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the present disclosure provides a method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (VI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the present disclosure provides a method of preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (VI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. [0412] In some embodiments, the present disclosure provides a method of treating or preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (VII), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the present disclosure provides a method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (VII), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the present disclosure provides a method of preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (VII), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
U1197.70243WO00 123/193 #13587456v2 [0413] In some embodiments, the present disclosure provides a method of treating or preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (VIII), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the present disclosure provides a method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (VIII), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the present disclosure provides a method of preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (VIII), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. [0414] In some embodiments, the present disclosure provides a method of treating or preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (IX), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the present disclosure provides a method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (IX), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the present disclosure provides a method of preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (IX), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. [0415] In some embodiments, the present disclosure provides a method of treating or preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (X), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the present disclosure provides a method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (X), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the present disclosure provides a method of preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (X), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. [0416] In some embodiments, the present disclosure provides a method of treating or preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (XI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the present
U1197.70243WO00 124/193 #13587456v2 disclosure provides a method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (XI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the present disclosure provides a method of preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (XI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. [0417] In another aspect, the present disclosure provides a provided compound or provided composition for use in the treatment or prevention of a disease or disorder. In some embodiments, the present disclosure provides a provided compound or provided composition for use in the treatment of a disease or disorder. In some embodiments, the present disclosure provides a provided compound or provided composition for use in the prevention of a disease or disorder. [0418] In another aspect, the present disclosure provides a compound of Formulae (I), (I′), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), or (XI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, for use in the treatment or prevention of a disease or disorder. In some embodiments, the present disclosure provides a compound of Formulae (I), (I′), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), or (XI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, for use in the treatment of a disease or disorder. In some embodiments, the present disclosure provides a compound of Formulae (I), (I′), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), or (XI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, for use in the prevention of a disease or disorder. [0419] In another aspect, the present disclosure provides a provided compound or provided composition for use in the manufacture of a medicament for the treatment or prevention of a disease or disorder. In some embodiments, the present disclosure provides a provided compound or provided composition for use in the manufacture of a medicament for the treatment of a disease or disorder. In some embodiments, the present disclosure provides a provided compound or provided composition for use in the manufacture of a medicament for the prevention of a disease or disorder. In another aspect, the present disclosure provides a compound of Formulae (I), (I′), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), or (XI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, for use in the manufacture of a medicament for the treatment or prevention of a disease or disorder. In some embodiments, the present disclosure provides a compound of Formulae (I), (I′), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), or (XI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, for use in the manufacture of a medicament for the treatment of a disease or disorder. In some embodiments, the present disclosure provides a compound of Formulae (I), (I′), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), or (XI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, for use in the manufacture of a medicament for the prevention of a disease or disorder.
U1197.70243WO00 125/193 #13587456v2 [0420] In some embodiments, the method of treating or preventing a disease or disorder comprises modulating a target protein. In some embodiments, modulating the target protein comprises inhibition of the target protein. In some embodiments, modulating the target protein comprises agonism of the target protein. In some embodiments, modulating the target protein comprises antagonism of the target protein. [0421] In some embodiments, the disease or disorder is associated with the target protein. In some embodiments, the disease or disorder is associated with signal transducer and activator of transcription 3 (STAT3), Toll-like receptor 7 (TLR7), indoleamine 2,3-dioxygenase (IDO), nitric oxide synthase (NOS), arginase, programmed death-ligand 1 (PD-L1), protein kinase B, cluster of differentiation 73 (CD73), or stimulator of interferon genes (STING). In some embodiments, the disease or disorder is associated with signal transducer and activator of transcription 3 (STAT3). In some embodiments, the disease or disorder is associated with Toll-like receptor 7 (TLR7). In some embodiments, the disease or disorder is associated with indoleamine 2,3-dioxygenase (IDO). In some embodiments, the disease or disorder is associated with nitric oxide synthase (NOS). In some embodiments, the disease or disorder is associated with arginase. In some embodiments, the disease or disorder is associated with programmed death-ligand 1 (PD-L1). In some embodiments, the disease or disorder is associated with protein kinase B. In some embodiments, the disease or disorder is associated with cluster of differentiation 73 (CD73). In some embodiments, the disease or disorder is associated with stimulator of interferon genes (STING). [0422] In some embodiments, the disease or disorder is a genetic disease, proliferative disease, hematological disease, neurological disease, liver disease, spleen disease, lung disease, painful condition, psychiatric disorder, musculoskeletal disease, a metabolic disorder, inflammatory disease, autoimmune disease, or microbial infection. In some embodiments, the disease or disorder is a proliferative disease. In some embodiments, the proliferative disease is cancer (e.g., a solid tumor (e.g., brain tumor (e.g., glioma, glioblastoma), pancreatic cancer, ovarian cancer)). In some embodiments, the proliferative disease comprises a solid tumor (e.g., brain tumor (e.g., glioma, glioblastoma), pancreatic cancer, ovarian cancer). In some embodiments, the proliferative disease is a solid tumor (e.g., brain tumor (e.g., glioma, glioblastoma), pancreatic cancer, ovarian cancer). In some embodiments, the proliferative disease is a brain tumor (e.g., glioma, glioblastoma), pancreatic cancer, or ovarian cancer. In some embodiments, the proliferative disease is a brain tumor (e.g., glioma, glioblastoma). In some embodiments, the proliferative disease is glioma. In some embodiments, the proliferative disease is glioblastoma. In some embodiments, the proliferative disease is pancreatic cancer. In some embodiments, the proliferative disease is ovarian cancer. [0423] In some embodiments, the disease or disorder is a liver disease or a spleen disease. In some embodiments, the disease or disorder is a liver disease. In some embodiments, the disease or disorder is a spleen disease.
U1197.70243WO00 126/193 #13587456v2 [0424] In some embodiments, the disease or disorder is a microbial infection (e.g., sepsis). In some embodiments, the disease or disorder is sepsis. [0425] In another aspect, the present disclosure provides a method of modulating a target protein, the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a provided compound or a provided composition. [0426] In another aspect, the present disclosure provides a method of modulating a target protein, the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formulae (I), (I′), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), or (XI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. [0427] In some embodiments, modulating the target protein comprises inhibition of the target protein. In some embodiments, modulating the target protein comprises agonism of the target protein. In some embodiments, modulating the target protein comprises antagonism of the target protein. [0428] In some embodiments, the target protein is signal transducer and activator of transcription 3 (STAT3), Toll-like receptor 7 (TLR7), indoleamine 2,3-dioxygenase (IDO), nitric oxide synthase (NOS), arginase, programmed death-ligand 1 (PD-L1), protein kinase B, cluster of differentiation 73 (CD73), or stimulator of interferon genes (STING). In some embodiments, the target protein is signal transducer and activator of transcription 3 (STAT3). In some embodiments, the target protein is Toll- like receptor 7 (TLR7). In some embodiments, the target protein is indoleamine 2,3-dioxygenase (IDO). In some embodiments, the target protein is nitric oxide synthase (NOS). In some embodiments, the target protein is arginase. In some embodiments, the target protein is programmed death-ligand 1 (PD-L1). In some embodiments, the target protein is protein kinase B. In some embodiments, the target protein is cluster of differentiation 73 (CD73). In some embodiments, the target protein is stimulator of interferon genes (STING). [0429] In some embodiments, the present disclosure provides a method of modulating a target protein, the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the method comprises inhibition of STAT3, agonism of TLR7, inhibition of IDO, inhibition of NOS, inhibition of arginase, inhibition of PD-L1, inhibition of protein kinase B, inhibition of CD73, or agonism of STING. In some embodiments, the method comprises inhibition of STAT3. In some embodiments, the method comprises agonism of TLR7. In some embodiments, the method comprises inhibition of IDO. In some embodiments, the method comprises inhibition of NOS. In some embodiments, the method comprises inhibition of arginase. In some embodiments, the method comprises inhibition of PD-L1. In some embodiments, the method comprises inhibition of protein kinase B. In some embodiments, the method comprises inhibition of CD73. In some embodiments, the method comprises agonism of STING.
U1197.70243WO00 127/193 #13587456v2 [0430] In some embodiments, the present disclosure provides a method of modulating a target protein, the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (I′), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the method comprises inhibition of STAT3, agonism of TLR7, inhibition of IDO, inhibition of NOS, inhibition of arginase, inhibition of PD-L1, inhibition of protein kinase B, inhibition of CD73, or agonism of STING. In some embodiments, the method comprises inhibition of STAT3. In some embodiments, the method comprises agonism of TLR7. In some embodiments, the method comprises inhibition of IDO. In some embodiments, the method comprises inhibition of NOS. In some embodiments, the method comprises inhibition of arginase. In some embodiments, the method comprises inhibition of PD-L1. In some embodiments, the method comprises inhibition of protein kinase B. In some embodiments, the method comprises inhibition of CD73. In some embodiments, the method comprises agonism of STING. [0431] In some embodiments, the present disclosure provides a method of modulating a target protein, the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (II), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the target protein is STAT3. In some embodiments, the method comprises inhibition of STAT3. [0432] In some embodiments, the present disclosure provides a method of modulating a target protein, the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (III), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the target protein is TLR7. In some embodiments, the method comprises agonism of TLR7. [0433] In some embodiments, the present disclosure provides a method of modulating a target protein, the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (IV), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the target protein is IDO. In some embodiments, the method comprises inhibition of IDO. [0434] In some embodiments, the present disclosure provides a method of modulating a target protein, the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (V), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the target protein is NOS. In some embodiments, the method comprises inhibition of NOS. [0435] In some embodiments, the present disclosure provides a method of modulating a target protein, the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (VI), or a pharmaceutically
U1197.70243WO00 128/193 #13587456v2 acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the target protein is arginase. In some embodiments, the method comprises inhibition of arginase. [0436] In some embodiments, the present disclosure provides a method of modulating a target protein, the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (VII), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the target protein is PD-L1. In some embodiments, the method comprises inhibition of PD-L1. [0437] In some embodiments, the present disclosure provides a method of modulating a target protein, the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (VIII), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the target protein is protein kinase B. In some embodiments, the method comprises inhibition of protein kinase B. [0438] In some embodiments, the present disclosure provides a method of modulating a target protein, the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (IX), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the target protein is CD73. In some embodiments, the method comprises inhibition of CD73. [0439] In some embodiments, the present disclosure provides a method of modulating a target protein, the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (X), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the target protein is STING. In some embodiments, the method comprises agonism of STING. [0440] In some embodiments, the present disclosure provides a method of modulating a target protein, the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (XI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the target protein is STAT3. In some embodiments, the method comprises inhibition of STAT3. [0441] In another aspect, the present disclosure provides a method of modulating signal transducer and activator of transcription 3 (STAT3), the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (II), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the modulating STAT3 comprises inhibition of STAT3. [0442] In another aspect, the present disclosure provides a method of modulating Toll-like receptor 7 (TLR7), the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (III), or a pharmaceutically
U1197.70243WO00 129/193 #13587456v2 acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the modulating TLR7 comprises agonism of TLR7. [0443] In another aspect, the present disclosure provides a method of modulating indoleamine 2,3- dioxygenase (IDO), the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (IV), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the modulating IDO comprises inhibition of IDO. [0444] In another aspect, the present disclosure provides a method of modulating nitric oxide synthase (NOS), the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (V), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the modulating NOS comprises inhibition of NOS. [0445] In another aspect, the present disclosure provides a method of modulating arginase, the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (VI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the modulating arginase comprises inhibition of arginase. [0446] In another aspect, the present disclosure provides a method of modulating programmed death- ligand 1 (PD-L1), the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (VII), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the modulating PD-L1 comprises inhibition of PD-L1. [0447] In another aspect, the present disclosure provides a method of modulating protein kinase B, the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (VIII), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the modulating protein kinase B comprises inhibition of protein kinase B. [0448] In another aspect, the present disclosure provides a method of modulating cluster of differentiation 73 (CD73), the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (IX), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the modulating CD73 comprises inhibition of CD73. [0449] In another aspect, the present disclosure provides a method of modulating stimulator of interferon genes (STING), the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (X), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the modulating STING comprises agonism of STING.
U1197.70243WO00 130/193 #13587456v2 [0450] In another aspect, the present disclosure provides a method of modulating signal transducer and activator of transcription 3 (STAT3), the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of a compound of Formula (XI), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the modulating STAT3 comprises inhibition of STAT3. [0451] In some embodiments, the administration is by injection. [0452] In some embodiments, the subject is an animal. The animal may be of either sex and may be at any stage of development. In some embodiments, the subject described herein is a human. In some embodiments, the subject is a non-human animal. In some embodiments, the subject is a mammal. In some embodiments, the subject is a non-human mammal. In some embodiments, the subject is a domesticated animal, such as a dog, cat, cow, pig, horse, sheep, or goat. In some embodiments, the subject is a companion animal, such as a dog or cat. In some embodiments, the subject is a livestock animal, such as a cow, pig, horse, sheep, or goat. In some embodiments, the subject is a zoo animal. In another embodiment, the subject is a research animal, such as a rodent (e.g., mouse, rat), dog, pig, or non-human primate. In some embodiments, the animal is a genetically engineered animal. In some embodiments, the animal is a transgenic animal (e.g., transgenic mice and transgenic pigs). In some embodiments, the subject is a fish or reptile. Methods of Preparation [0453] In another aspect, the present disclosure provides a method of preparing a compound of or a pharmaceutically acceptable salt thereof, the method comprising: providing a reaction vessel; adding to the reaction vessel a compound of Formula (XIII-a): (XIII-a), or a salt thereof; adding to the reaction vessel a compound of Formula (XIV-a):
U1197.70243WO00 131/193 #13587456v2 or a salt thereof; and obtaining the compound of Formula (II-c), or pharmaceutically acceptable salt thereof; wherein: Zn is an nth generation polyamidoamine (PAMAM) dendrimer; n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; each instance of RA is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; each of p and p1 is independently an integer between 1 and 2(n+2), inclusive; each of q and q1 is independently an integer between 0 and 2(n+2)-1, inclusive; the sum of p and q is 2(n+2); the sum of p1 and q1 is 2(n+2); p is less than or equal to p1; and q is greater than or equal to q1. [0454] In another aspect, the present disclosure provides a method of preparing a compound of Formula (II-d): or a pharmaceutically acceptable salt thereof, the method comprising: providing a reaction vessel; adding to the reaction vessel a compound of Formula (XIII-b): or a salt thereof;
U1197.70243WO00 132/193 #13587456v2 adding to the reaction vessel a compound of Formula (XIV-b): or a salt thereof; and obtaining the compound of Formula (II-d), or pharmaceutically acceptable salt thereof; wherein: Zn is an nth generation polyamidoamine (PAMAM) dendrimer; n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; each instance of RA is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; each of p and p2 is independently an integer between 1 and 2(n+2), inclusive; each of q and q2 is independently an integer between 0 and 2(n+2)-1, inclusive; the sum of p and q is 2(n+2); the sum of p2 and q2 is 2(n+2); p is less than or equal to p2; and q is greater than or equal to q2. [0455] In another aspect, the present disclosure provides a method of preparing a compound of Formula (XI-d): or a pharmaceutically acceptable salt thereof, the method comprising: providing a reaction vessel; adding to the reaction vessel a compound of Formula (XIII-b):
U1197.70243WO00 133/193 #13587456v2 or a salt thereof; adding to the reaction vessel a compound of Formula (XIV-c): or a salt thereof; and obtaining the compound of Formula (XI-d), or pharmaceutically acceptable salt thereof; wherein: Zn is an nth generation polyamidoamine (PAMAM) dendrimer; n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; each instance of RA is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; each of p and p2 is independently an integer between 1 and 2(n+2), inclusive; each of q and q2 is independently an integer between 0 and 2(n+2)-1, inclusive; the sum of p and q is 2(n+2); the sum of p2 and q2 is 2(n+2); p is less than or equal to p2; and q is greater than or equal to q2. [0456] In another aspect, the present disclosure provides a compound, or a pharmaceutically acceptable salt thereof, made by the method comprising: providing a reaction vessel; adding to the reaction vessel a compound of Formula (XIII-a): (XIII-a), or a salt thereof; adding to the reaction vessel a compound of Formula (XIV-a):
U1197.70243WO00 134/193 #13587456v2 or a salt thereof; and obtaining the compound, or pharmaceutically acceptable salt thereof; wherein: Zn is an nth generation polyamidoamine (PAMAM) dendrimer; n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; each instance of RA is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; p1 is an integer between 1 and 2(n+2), inclusive; q1 is an integer between 0 and 2(n+2)-1, inclusive; and the sum of p1 and q1 is 2(n+2). [0457] In some embodiments, the method is according to conditions provided in the Examples. [0458] In another aspect, the present disclosure provides a compound, or a pharmaceutically acceptable salt thereof, made by the method comprising: providing a reaction vessel; adding to the reaction vessel a compound of Formula (XIII-b): or a salt thereof; adding to the reaction vessel a compound of Formula (XIV-b): or a salt thereof; and obtaining the compound, or pharmaceutically acceptable salt thereof; wherein: Zn is an nth generation polyamidoamine (PAMAM) dendrimer; n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10;
U1197.70243WO00 135/193 #13587456v2 each instance of RA is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; p2 is an integer between 1 and 2(n+2), inclusive; q2 is an integer between 0 and 2(n+2)-1, inclusive; and the sum of p2 and q2 is 2(n+2). [0459] In some embodiments, the method is according to conditions provided in the Examples. [0460] In another aspect, the present disclosure provides a compound, or a pharmaceutically acceptable salt thereof, made by the method comprising: providing a reaction vessel; adding to the reaction vessel a compound of Formula (XIII-b): or a salt thereof; adding to the reaction vessel a compound of Formula (XIV-c): or a salt thereof; and obtaining the compound, or pharmaceutically acceptable salt thereof; wherein: Zn is an nth generation polyamidoamine (PAMAM) dendrimer; n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; each instance of RA is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; p2 is an integer between 1 and 2(n+2), inclusive; q2 is an integer between 0 and 2(n+2)-1, inclusive; and the sum of p2 and q2 is 2(n+2). [0461] In some embodiments, the method is according to conditions provided in the Examples.
U1197.70243WO00 136/193 #13587456v2 EXAMPLES [0462] In order that the present disclosure may be more fully understood, the following examples are set forth. The synthetic and biological examples described in this application are offered to illustrate the compounds, pharmaceutical compositions, and methods provided herein and are not to be construed in any way as limiting in their scope. Example 1: Materials and Methods for Dendrimer Targeting and Uptake Studies [0463] Generation of Cy5-labeled dendrimers. The G6 PAMAM hydroxyl (OH), amine (NH2), and succinamic acid (SA) surface dendrimers were purchased from (Dendritech, Inc). The NH2 surface dendrimer was used as such after the evaporation of methanol from the stock solution. The OH and SA surface dendrimers were further functionalized with amine terminals to conjugate Cy5 mono NHS ester (Cytiva). For the Cy5-labeling of OH dendrimers: Step 1: Fmoc-GABA-OH (Sigma- Aldrich) was coupled with G6-OH using PyBOP (Merck) as a coupling reagent to produce an intermediate with protecting group Fmoc. Step 2: The Fmoc protecting group was removed using piperidine (Sigma-Aldrich) – DMF (Sigma-Aldrich) mixture to produce bi-functional dendrimers. The crude was purified by dialyzing (membrane cutoff = 12–14 kDa) against DMF (Sigma-Aldrich) for 24 h by changing the DMF every 8 h. Step 3: The conjugation of Cy5 mono NHS ester was carried out in the presence of borate buffer (pH 8.5) with pure bifunctional dendrimer to produce G6-OH-Cy5 conjugate. Similarly, amine surface G6 dendrimer was labeled with Cy5 using borate buffer (pH 8.5). For the Cy5-labeling of SA dendrimers, in step 1, G6 succinamic acid surface dendrimer was coupled with N-Fmoc 1,5-diaminobutane hydrobromide (Sigma-Aldrich) using EDC.HCl (Sigma-Aldrich) as a coupling reagent. In step 2, the successful deprotection of Fmoc using piperidine (Sigma-Aldrich) DMF (Sigma-Aldrich) mixture resulted in bi-functional dendrimers. The Cy5-labeling of bi-functional dendrimers under DMSO (Sigma-Aldrich) and DIEA (Sigma-Aldrich) produced G6-succinamic acid- Cy5 conjugate in step 3. The synthesized G6 PAMAM-Cy5 conjugates are in good agreement with the reported literature data28. [0464] Cell lines. The murine KR158 and GL261 glioma cell lines were received from Dr. Jeffrey Harrison18. The GL261 glioma cells were cultured in Dulbecco’s Modified Eagles Medium (DMEM) (Invitrogen) with 1% penicillin-streptomycin (Invitrogen) and 10% fetal bovine serum (FBS) (Thermo Scientific). Cells were expanded in T75 flasks (Falcon) in a humidified incubator (Thermo Scientific) at 37°C with 5% CO2. All cell lines tested negative for mycoplasma based on DNA-based PCR tests. [0465] M-MDSC induction and culture. Induction of M-MDSCs from transgenic CCR2RFP/WT/CX3CR1GFP/WT bone marrow cells was adapted from previously published work using wildtype C57BL/6 mice29. Bone marrow cells collected from the femur were seeded at a density of 1 x 106 cells/mL in KR158 cell-conditioned culture media (50% v/v KR158 conditioned media + 50% RPMI-1640 (Gibco) + 10% FBS (Corning) + 1% penicillin-streptomycin (Corning), + 1% GlutaMax (Gibco) + 1% Non-Essential Amino Acids (Gibco), 0.22µm sterile bottle-top filtered). Cells were
U1197.70243WO00 137/193 #13587456v2 cultured for five days. At the endpoint, suspension cells were collected from the supernatant and adherent cells by scraper (Fisher Scientific) after 15-minute incubation at 37°C, 5% CO2 with enzyme-free cell dissociation buffer (Gibco). Flasks were twice-washed using 10-25mL FACS buffer (10% FBS + 1× HBSS) and all cells were collected by centrifugation (500 x g for 5 minutes at 4°C). Cells were collectively resuspended in a 50mL sterile conical (Falcon) in FACS buffer and counted using trypan blue exclusion method. Cells were then analyzed in triplicate biological repeats by flow cytometry as described previously (Flow Cytometry Analysis). [0466] Characterization of dendrimer size distribution and ζ-potential and dendrimer-serum protein interaction. The physicochemical properties (size distribution and ζ-potential (zeta potential)) of NH2, OH, and SA dendrimers were characterized using Litesizer 500 (Anton Paar Instruments) at 25°C. To measure the hydrodynamic radius based on dynamic light scattering (DLS), all dendrimers with different terminal groups were dissolved in 1×PBS buffer (pH = 7.4) at 1mg/mL. The dendrimer solutions were filtered through a 0.22µm 13mm polyether sulfone (PES) syringe filter (Cytiva) before the size measurement within a 1 mL cuvette (Sarstedt). To measure the ζ-potential, dendrimers were diluted in 1×PBS buffer (pH = 7.4) at 0.3mg/mL. The dendrimer solutions were also filtered before the ζ-potential measurement within an omega cuvette (Anton Paar). To assess the interactions between dendrimers and serum proteins, dendrimers were incubated with mouse serum from C57BL/6j mice (in-house generated) for 30min at 37°C at a concentration of 0.86 mg/mL to allow the formation of the dendrimer-serum protein complex. The dendrimer-serum protein or the serum protein solutions per se were then diluted in 1×PBS buffer (pH = 7.4) to reach a final concentration of 0.3mg/mL before assessment. [0467] Mice and in vivo tumor models. Transgenic CCR2RFP/WT/CX3CR1GFP/WT C57BL/6 mice were bred in-house at the University of Florida animal facility. CCR2 RFP/WT/CX3CR1GFP/WT were generated by cross-breeding CCR2 deficient mice (CCR2 RFP/RFP[B6.129(Cg)-CCR2tm2.1lfv/J]) and CX3CR1 deficient mice (CX3CR1GFP/GFP[B6.129P-CX3CR1tm1Litt/J]). Wildtype C57BL/6 mice were purchased from The Jackson Laboratory. All procedures involving animal housing, care, and surgical procedures were following the guidelines of the University of Florida Institutional Animal Care and Use Committee. [0468] Mouse models of Glioma. To generate an orthotopic model of GL261 and KR158 murine gliomas, mice were anesthetized by controlled isoflurane inhalation, and their heads were shaved before intravenous analgesia administration. Surgical sites were prepared using 2-3mm incisions at the midline of the skull. Stereotactic injection of 2µL at 1µL/min (5.0 x 104) cells suspended in methylcellulose was performed at 2mm lateral from the bregma using a Hamilton syringe autonomously controlled by a micro-fluidic injection apparatus (Stoelting). Post-injection the dermal incision was closed via suture and bone wax application. Animals were placed on a cage warmer for post-surgical monitoring. For in vivo studies of dendrimer uptake and distribution, Mice received tail
U1197.70243WO00 138/193 #13587456v2 vein injections of SA and OH dendrimers (50mg/kg) and NH2 dendrimers (10mg/kg). Cy5-labeled dendrimers were suspended in 100-200µL saline and filtered with 0.22µm 13mm polyether sulfone (PES) filters. [0469] Flow cytometry sample preparation and analysis. Cells obtained from brain tumor, bone marrow (femur), spleen, and blood were analyzed by flow cytometry. Mouse blood was collected from the chest cavity post right atrium lancing using a 1mL syringe coated with 0.5M EDTA (Invitrogen). Approximately 200µL of blood was transferred to a 1.5mL microcentrifuge (Fisher Scientific) tube containing 100µL 0.5M EDTA. Whole blood was centrifuged at 21°C, 380 x g for 5 minutes and the plasma was collected and stored at -80°C in 1.5mL microcentrifuge tubes. Before the collection of other organs, systemic perfusion was then performed by needle insertion into the left ventricle and flush with 20mL 1× PBS (Gibco) using a 10mL syringe (BD) and 25G butterfly infusion set (Exel). Brains were removed by sagittal and coronal partitioning of the skull using surgical scissors and transferred to a microscopy slide for tumor excision. To generate a single-cell suspension for analysis, tumor tissue was minced using a regular single-edge razor blade until a viscous suspension of cells was generated. Cells were then transferred to a 50mL conical (Falcon) filled with Accumax dissociation buffer (Innovative Cell Technologies) and incubated in a 37°C water bath for 5 minutes. Suspensions were then oscillated through a 1mL single-channel pipet tip for 40 cycles and strained through a 40µm strainer into a 50mL conical, followed by dilution with 5mL FACS buffer (10% FBS, 1× HBSS). Cells were collected by centrifugation at 380 x g for 5 minutes at 4°C, followed by resuspension in 70% v/v Percoll Solution (70% Percoll, 1% 1× PBS in RPMI-1640). Using a 5mL syringe and 3-inch 18G needle, the 70% Percoll suspension of tumor cells was injected below a layer of 37% v/v Percoll (4mL, 37% Percoll, and 1% 1× PBS in RPMI-1640) (Gibco) in a 15 mL conical. Samples were subsequently centrifuged at 500 x g for 30 minutes at 21°C (level 1 acceleration, level 0 deacceleration). The resulting tumor cell interface between Percoll layers was removed (1mL) by a single channel pipet and transferred to a 1.5mL microcentrifuge tube. Cells were centrifuged at 500 ×g for 5 minutes at 4°C and washed and resuspended with ice-cold FACS Buffer. Femurs were harvested and ends clipped with dissecting scissors after connective tissues were removed. The isolated femurs were placed in 0.5mL microcentrifuge tubes with an 18G needle pierced bottom, cap removed, and tube nested within a secondary 1.5 microcentrifuge tube containing 100µL ice cold ACK lysis buffer (Gibco). Microcentrifuge tubes with femurs were centrifuged at 5,700 RPM for 20 seconds at 21°C to capture bone marrow. Spleens were excised and transferred to a petri dish on ice for mincing using a regular single-edge razor blade (Personna) after injection with 1mL of ice-cold 1× HBSS (Gibco) using a 3-inch 18G needle (Air-Tite) and 5mL syringe (BD). Dispersed tissues were aspirated into a 5mL syringe via a 3-inch 18G needle and transferred to a 15mL conical.5mL of ice-cold 1× HBSS (Gibco) was added, and cells were mechanically dissociated by the oscillation of the volume through the syringe and needle for 20 cycles. The resulting splenocyte
U1197.70243WO00 139/193 #13587456v2 suspension was centrifuged at 380 x g for 5 minutes at 4°C.1mL ice cold ACK Lysis Buffer (Gibco) was added to bone marrow cells, leukocytes, and splenocyte to resuspend post centrifugation for 5 minutes and subsequently diluted with 5mL ice-cold FACS Buffer (10% FBS in 1× HBSS) then strained through a 40µm cell strainer. Cells from each tissue were isolated by centrifugation at 380 x g for 5 minutes at 4°C. To remove all visibly present red blood cells, leukocytes were repeatedly cycled, up to four additional times, through ACK lysis buffer (Gibco) and FACS buffer wash as previously described. Viability was manually determined by cell count using a standard trypan blue (Corning) exclusion method. [0470] Single-cell suspensions were prepared as described in the above sections. Samples were stained with viability dye (Violet, Invitrogen) in 1× PBS pH 7.4 (Gibco) at RT protected from light for 15 minutes. Cells were resuspended and washed with FACS buffer (10% FBS, 1× HBSS) and stored on ice until analysis. Samples were analyzed via a single flow cytometry tube on a Sony Spectral Analyzer (SP6800). A multi-color reference control panel consisting of transgenic single color CCR2RFP/WT and CX3CR1GFP/WT bone marrow cells, viability dye–violet (Thermo Scientific), and Cy5-positive wildtype C57BL/6 bone marrow cell suspensions was utilized to unmix panels as appropriate. Raw data was subsequently analyzed and graphically illustrated using FlowJo software (BD Biosciences). [0471] In vitro dendrimer uptake study. Transgenic CCR2RFP/WT/CX3CR1GFP/WT bone marrow cells were derived into M-MDSCs ex vivo as described herein (CCR2RFP+/CX3CR1GFP+). Cells were washed with FACS buffer (10% FBS, 1× HBSS) and resuspended in serum-free 1× HBSS (Gibco). Viability and concentration were determined by trypan blue exclusion. Cells were adjusted to a concentration of ~1 × 106/mL in 1.5mL microcentrifuge tubes (Fisher Scientific) and centrifuged at 500 x g for 5 minutes at 4°C. Cells were resuspended in 400µL of dissolved dendrimer solution at a concentration range from 1-100µg/mL in 1× HBSS or 1× PBS at room temperature and incubated for 30 minutes protected from light. Samples were then washed in 1× HBSS, stained for viability, and resuspended with FACS buffer in three technical repeats and analyzed via Spectral Flow Cytometry as described herein. To determine how dendrimer-associated serum proteins affect their interaction with M-MDSCs in vitro, dendrimer stock solutions were prepared by fully solubilizing dendrimers in 1× PBS pH 7.4 (Gibco), followed with filtration through a 13mm 0.22µm PES syringe filter (Cytiva). Solutions were diluted at room temperature to concertation of 0.86 mg/mL in either competent or heat-inactivated (60°C for 30 minutes) sex pooled, complement preserved, C57BL/6 murine serum (Charles River) in 1.5mL microcentrifuge tubes. Dendrimer-serum stock solutions were then incubated at 37°C for 30 minutes and brought to room temperature before co-incubation with cells at escalating doses (5-100µ/mL) in three biological repeats. The dendrimer uptake as indicated by Mean Fluorescence Intensity (MFI) was subjected to flow cytometry analysis by gating out the CCR2RFP+/CX3CR1GFP+ (M-MDSC) population.
U1197.70243WO00 140/193 #13587456v2 [0472] Immunofluorescence study. To determine the biodistribution and the cell uptake of dendrimers, Cy5-labeled dendrimers filtered were injected via tail vein into transgenic CCR2RFP/WT/CX3CR1GFP/WT or wildtype C57BL/6 mice at tolerable doses (50 mg/kg for SA, OH dendrimers and 10mg/kg for NH2 dendrimers). Euthanized animals were systemically perfused with 20mL 1× PBS (Gibco) and 20mL 4% w/v paraformaldehyde (PFA) buffered solution. (Thermo Scientific) using a 50ml syringe and 25G butterfly needle infusion set. Brain (tumors), spleens, and femurs were excised, and connective tissues were removed prior to transfer to 5 mL of 4 % w/v PFA at 4°C for 1 hour (brains and spleens) or 72 hours (femurs) at 2–8°C. Brains (tumors) and spleens were then transferred to a 30% w/v sucrose (Fisher Scientific) in water (Corning) solution for ≥ 24 hours in 15mL conical tubes stored at 2–8°C. Femurs were subsequently transferred to 5 mL of decalcification solution (20% EDTA, 10N NaOH, pH 7.4) for 4 days at 2-8°C and then a 30% w/v sucrose solution for 24 hours at 2-8°C. All tissues were embedded in optimal cutting temperature compound (Fisher Scientific) and cryo-sectioned at 10μm or 30μm thick sections at -25°C. Sections were prepared by addition to microscopy slides (Fisher Scientific), washed for 3 repetitive cycles with cold 1× Dulbecco’s Phosphate Buffered Saline (DPBS) in a staining dish. For vascular endothelial cell staining of brain tissues, an anti-mouse CD31-Spark YG 570 labeled mAb (BioLegend) was added to hydrophobic pen (Vector Laboratories) encircled sections at a concentration of 10 µg/mL. Slides were then mounted with Vectashield anti-fade mounting medium with DAPI stain (Vector Laboratories) and coverslip. Slides were sealed with CoverGrip sealant and subsequently stored at 2- 8°C protected from light in the staining tray. Sections were analyzed at high magnification using an inverted Nikon A1R confocal microscope. Widefield fluorescent images were generated using a Keyence BZ-X800 or Nikon Ti-E for fluorescence microscopy. Widefield fluorescence microscopy images were processed using Nikon Elements software v5.21 and confocal fluorescence microscopy using Fiji v2.9.0. [0473] Quantification of dendrimer concentration in plasma. Blood was collected from euthanized CCR2RFP/WT/CX3CR1GFP/WT mice bearing 3–4-week KR158 or GL261 gliomas at 24- or 72-hours post-dendrimer administration (tail vein) as previously described herein. Plasma samples were thawed from -80°C storage to room temperature and diluted 1:4 with 1× PBS (Gibco). Samples were then transferred to a 96-well clear bottom black plate (Thermo Scientific) and analyzed for absolute fluorescence intensity from Cy5 (635/675 (ex/em), integration: 400ms, read height: 3.0mm) using a Molecular Devices SpectraMax iD3 Multi-Mode Microplate Reader. Samples were plotted against a standard curve of Cy5 in murine serum and interpolated post-transgenic murine plasma background subtraction. The percentage of the injected dose was calculated by dilution factors × estimated dendrimer concentration × plasma volume (estimated to be 1.8 mL/mouse) / total injected dose. Samples were evaluated by 3 technical repeats.
U1197.70243WO00 141/193 #13587456v2 [0474] Data reporting and statistical analysis. Each in vitro assay was performed using a minimum of three technical repeats or three biological repeats. All data was processed and graphed using GraphPad Prism v10.1.1 displaying average, standard deviations, error, and statistical p-values by one or two-way ANOVA as appropriate per data set. Each in vivo assay was performed using a minimum of n=6 mice based on the median of a group comparison using a one- or two-way ANOVA between the calculated min and max degrees of freedom ((DF=k(n-1) n=sample size, k=number of groups). Representative immunohistochemistry n=1 (IHC) and microscopy tissue samples were displayed. Example 2: Dendrimer Targeting and Uptake Studies [0475] Introduction. The focus of nanoparticles in vivo trafficking has been mostly on their tissue- level biodistribution and clearance. Recent progress in the nanomedicine field suggests that the targeting of nanoparticles to immune cells can be used to modulate the immune response and enhance therapeutic delivery to the diseased tissue. In the presence of tumor lesions, monocytic-myeloid- derived suppressor cells (M-MDSCs) expand significantly in the bone marrow, egress into peripheral blood, and traffic solid tumor, where they help maintain an immuno-suppressive tumor microenvironment. [0476] In the present disclosure, the interaction between PAMAM dendrimers and M-MDSCs was quantitatively examined in two murine models of glioblastoma, by examining the cell-level biodistribution kinetics of the systemically injected dendrimers. M-MDSCs in the tumor and lymphoid organs (bone marrow, spleen, peripheral blood, and tumor) can efficiently endocytose hydroxyl dendrimers. The trafficking of M-MDSCs from the bone marrow to the tumor contributed to the deposition of hydroxyl dendrimers in the tumor. M-MDSCs showed different capacities of endocytosing dendrimers of different functionalities in vivo. In the tumor, M-MDSCs and microglia showed a high capacity of taking up hydroxyl dendrimers and these two cellular compartments accounted for more than half the amount of the hydroxyl dendrimer deposition in the tumor. This differential uptake and targeting was mediated by the unique serum proteins associated with each dendrimer surface functionality. [0477] M-MDSCs are recruited to the tumor from bone marrow. To determine how dendrimers interact with the M-MDSCs and other immune cells in vivo, the profiles of M-MDSCs and other infiltrative immune cells were first characterized in a GL261 mouse glioma model. This model well- recapitulates the histology of glioma and has been extensively used to test the therapeutic responses in the literatures30. Here, the GL261 glioma model was set up using CCR2RFP/WTCX3CR1GFP/WT transgenic mice, which allows the direct surveillance of the profile of infiltrative immune cells via the endogenously expressed red fluorescent protein (RFP) for chemokine receptor two (CCR2) and green fluorescent protein (GFP) for C-X3-C motif chemokine receptor 1 (CX3CR1)18, 31. When expressed jointly, these G-Protein Coupled Receptors (GPCRs) have been established as an equivalent biomarker for the M-MDSC cell subset, as defined by CD45+Ly6G-Ly6C+CD11b+ populations2, these
U1197.70243WO00 142/193 #13587456v2 cells have been shown to suppress both CD4+ and CD8+ T cells in mouse glioma model29. At 2-3 weeks after the tumor initiation, flow cytometry analysis of the M-MDSC population at the bone marrow was performed, which serves as the hematopoietic tissue of M-MDSCs; at the blood, which serves as the conduit to their destination; and at the spleen, which serves as a temporary reservoir. M- MDSCs accounted for 11.4±0.4% and 3.8±0.1% of total cells in the bone marrow and spleen of GL261 tumor-bearing mice (FIGs.1A-1B). CCR2 and its cognate receptors mediated M-MDSCs egress from bone marrow into peripheral blood29, in which the M-MDSCs comprised 8.5±1.5% of the blood leukocytes (FIGs.1A-1B); M-MDSCs infiltrated the glioma through peripheral blood, ultimately comprising 22.1±1.0% of the stromal cells in the GL261 tumor (FIG.1C). In the TME, M- MDSCs were shown as the RFP and GFP co-localized cells, as indicated by the arrows in FIG.1D. The CCR2RFP/WTCX3CR1GFP/WT transgenic mice also enabled profiling of other immune cell subsets in the glioma TME. Based on the published data18, the CCR2-/CX3CR1+ subsets (16.7±3.0%, FIG.1C, abbreviated as microglia) were CD45low/MHC+/F4/80+/CD11c-/CD11bmedium, likely representing the CNS tissue-resident microglia; the CCR2+/CX3CR1- subsets (4.4±2.0%, FIG.1C, abbreviated as CCR2+) were CD45+/MHCII+/F4/80-/CD11c-/CD11blow, likely representing other infiltrate myeloid cells originated outside of the CNS; the CCR2-/CX3CR1- subsets (26.4±4.9%, FIG.1C, abbreviated as other cells) were a collection of tumor cells and other tumor stroma cells. Finally, the CCR2- /CX3CR1meidum subsets accounted for 30.5±5.9% of tumor stromal cells (FIG.1C, abbreviated as CXCR1int). Given these cells were mostly CD45-positive18, it is possible that they infiltrated the glioma from outside of the brain. [0478] Tumor M-MDSCs show a high capacity of dendrimer uptake. Previous studies have established that in the presence of neuroinflammation/tumor lesions, systemically injected OH dendrimers can selectively localize in activated glial cells in a spectrum of central nervous system (CNS) disorders26-28. PAMAM OH dendrimer (Generation 6) was used as a model dendrimer to probe the dendrimer uptake capacity of different cell subsets within the glioma TME and the lymphoid organs such as bone marrow, spleen, and blood. Herein, Mice with established GL261 glioma were injected systemically with OH dendrimers at 50mg/kg – a dose that has been well-tolerated in vivo32. To track the dendrimer–cell interaction, the OH dendrimer was fluorescently labeled with a minimal amount of Cy5 dye (~5% by wt%)27. At 24 hours after injection, different cell subsets within the stroma of the GL261 tumor were isolated and were subjected to flow cytometry analyses for the Cy5 Mean Fluorescence Intensity (MFI) within each cell subset, which indicates the amount of dendrimer being endocytosed up by the cells. Tumor M-MDSCs and microglia showed a capacity for high dendrimer uptake (FIG.2A). Specifically, the MFI of tumor M-MDSCs =1061±535, which was significantly higher than CX3CR1int, CCR2+, and other cells (FIG.2B). This indicated that tumor M- MDSCs have a higher capacity for endocytosing OH dendrimers than other cell subsets within the GL261 tumor. The composition of all dendrimer-positive cells was evaluated within the GL261 tumor by gating out the dendrimer-positive populations from the whole tumor stroma cells. The
U1197.70243WO00 143/193 #13587456v2 compositions of the dendrimer-positive populations were then analyzed based on the CCR2 and CX3CR1 expression (FIG.2C). The majority of the dendrimer-positive cells were mostly distributed within 4 cellular compartments, i.e., M-MDSC (19.7±6.7%), microglia (25.1±3.4%), CX3CR1int (28.6±6.9%), and other cells (24.0±6.2%) (FIG.2D). CCR2+ compartment only accounted for 2.7±0.7% of dendrimer-positive cells, potentially due to their small numbers within the tumor stroma (~5%, FIG.1C). [0479] While the GL261 glioma model well-recapitulates the histology of glioma30, it is well- established that the GL261 glioma model, unlike human glioblastoma, is immunogenic30, 33, 34. Specifically, GL261 has high MHC I expression and a high tumor mutational load33 and responds well to checkpoint inhibitors34. The dendrimer interactions with M-MDSC and other immune infiltrative cells were next characterized in an immunosuppressive KR158 model with lower populations of infiltrative M-MDSC35, 36. When comparing the overall Cy5 MFI of all tumor stromal cells, the GL261 tumor showed 1.6-fold higher MFI (mean=946.8) than the KR158 model (mean=598.2), indicating a higher dendrimer deposition in the GL261 tumor than in the KR158 tumor. The different dendrimer deposition between the two glioma models was also reflected at the cellular level. The cell subsets within the GL261 tumor showed approximately 2–3-fold higher dendrimer uptake than the KR158 tumor (FIG.2F). When comparing the cellular composition of dendrimer-positive populations, around 53.4±6.0% of dendrimer-positive cells in the KR158 tumor were located in the microglia compartment, while only 6.4±0.8% and 12.1±2.6% of dendrimer-positive cells were located in the M-MDSC and CX3CR1int compartments respectively (FIG.2G). Although the KR158 tumor had lower dendrimer deposition and different compositions of dendrimer-positive cells compared to the GL261 tumor, the M-MDSCs and microglia in both tumor models showed higher dendrimer uptake than other cell subsets (FIG.2F, FIGs.6A-6C). In summary, the data based on two different glioma models indicated that the monocytic myeloid cells largely contributed to the tumor depositions of OH dendrimers. It was next evaluated whether the percentage of each cell subset within the tumor could correlate with the amount of dendrimer deposition in both tumor models. Analyses of the Pearson correlation coefficients showed that the percentage of tumor-infiltrative cells (M-MDSCs, Microglia, CX3CR1int, and CCR2+) generally had positive correlations with the amount of OH dendrimer depositions in both tumor models (FIG.2H). Specifically, M-MDSCs and CX3CR1int population showed good correlation (R> 0.2 or R>0.6) in both tumor models. However, the percentage of CCR2-/CX3CR1- subsets (other cells), which are a collection of tumor cells and other tumor stroma cells, showed a strong negative correlation with OH dendrimer deposition (R<0.6). [0480] The selective uptake of OH dendrimer by tumor-associated microglia/macrophages has been reported in previous studies26, 27. However, the ontogeny difference between CNS-resident microglia and bone marrow-derived macrophages has led to their different functions in cancer and different responses to macrophage-targeting therapeutics37, 38, indicating the importance of analyzing the cell-
U1197.70243WO00 144/193 #13587456v2 level biodistribution of nanotherapeutics. Both the M-MDSCs originated from bone marrow and the CNS-resident microglia showed a strong capacity for taking up OH dendrimers in this study. However, it is possible these cell subsets endocytose dendrimers through different mechanisms. For example, bone marrow-originated M-MDSCs and tumor-associated macrophages are more associated with phagocytosis and antigen-presentation, they tend to show higher density at perivascular niche than microglia, which display signatures associated with synaptic pruning39, 40. Differential dendrimer deposition was also observed between the GL261 and KR158 tumors. This difference might be associated with the different immunogenicity of GL261 and KR158 tumors30. The immunogenic GL261 tumor has a ‘hotter’ tumor milieu with more infiltrative immune cells than the KR158 tumor. Since the infiltrative myeloid cells within the tumor can contribute to the tumor-accumulation of nanoparticles41, 42, it is possible that the higher OH dendrimer deposition in the GL261 tumor was associated with the higher amount of infiltrative immune cells in the GL261 tumor. The KR158 tumor showed a mushroom-like crown on top of the brain, while the GL261 histology was more representative of the human glioma (FIG.7). The histological difference could also contribute to the differential dendrimer uptake. [0481] The trafficking kinetics of M-MDSC contribute to the dendrimer accumulation in the tumor. In the presence of tumor lesions, the production of M-MDSCs is accelerated in the bone marrow, from which these cells are directly recruited to the brain tumor through peripheral blood or indirectly from the spleen, which serves as the temporary reservoir of M-MDSCs18, 29, 31 (FIG.3A). To determine whether the trafficking kinetics of M-MDSCs affected the deposition of dendrimer in the brain tumor, the cellular uptake of OH dendrimers was quantified (MFI) by M-MDSCs located in bone marrow, spleen, peripheral blood, and tumor (GL261) at 24 hours after dendrimer injection (50mg/kg). Dendrimer uptake was observed in the M-MDSCs from all these tissues (FIG.3B). In the femur bone, OH dendrimers were mostly distributed in the red marrow, where the hematopoiesis led to the product of leukocytes (FIG.8A). In the spleen, OH dendrimers were mostly distributed in the red pulp (FIG.8B), where MDSCs are located43. The blood M-MDSCs showed the highest dendrimer uptake (MFI=1371±494), likely because blood M-MDSCs can directly access the dendrimers in the circulation without the limitation of any tissue barriers. The dendrimer uptake was further analyzed in different blood leukocytes (identified through the FSC- and SCC-based scattered plots). The dendrimer uptake was highest in granulocytes (MFI=1112±232), followed by monocytes (MFI=231.7±48) and lymphocytes (MFI=172.8±56) (FIG.3C and FIG.9A). Approximately 95% of granulocytes showed dendrimer uptake, compared to ~40% for monocytes and ~30% for lymphocytes (FIG.9B). Since M-MDSCs are constantly being recruited to the tumor in large amounts during tumor development, blood M-MDSCs may carry endocytosed dendrimer to the tumor while they infiltrate the tumor stroma. To test this hypothesis, the change of the dendrimer-positive M-MDSCs percentage was analyzed between the 24- and 72-hour window in two cohorts of mice. In bone
U1197.70243WO00 145/193 #13587456v2 marrow, there was a 50% decrease in the percentage of dendrimer-positive M-MDSCs within the 48- hour window (FIG.3D). This decrease in dendrimer-positive M-MDSCs could be caused by two factors. First, the emergency myelopoiesis in cancer leads to the accelerated generation of new M- MDSCs, which could dilute the dendrimer-positive populations; Second, the initial dendrimer- positive M-MDSCs could egress the bone marrow, further diluting the percentage of dendrimer- positive M-MDSC in the bone marrow. In the GL261 tumor, there was a 90% increase in the percentage of dendrimer-positive M-MDSCs (FIG.3D). This significant increase in dendrimer- positive M-MDSCs was likely caused by the recruitment of external dendrimer-positive M-MDSCs to the tumor milieu during the 48-hour window. It is less likely that tumor M-MDSCs could uptake more dendrimers within the 48–72 hours window, as further analysis of serum concentration showed that the amount of OH dendrimers decreased from 12.5±8.5% (total injected dose) at 24 hours to only 4.6±3.7% at 72 hours (FIG.3E). There was no significant change in dendrimer-positive populations in the blood and spleen M-MDSCs. [0482] For decades, the Enhanced Permeability and Retention (EPR) effect has been used as the guiding principle for designing tumor-targeting nanomedicine2. The accumulation of OH dendrimers in glioma was largely attributed to their neutral surface and ultra-small sizes (sub-10nm), which allowed them to efficiently cross the impaired blood-brain tumor barriers (BBTB) taken up by myeloid cells within the tumor through EPR effect and26, 27. However, it was not clear whether other complementary mechanism(s) could also contribute to the dendrimer accumulation in the tumor. Myeloid cells can modulate the pharmacokinetics, biodistribution, and efficacy of nanotherapeutics10, 11. Circulating myeloid cells, such as inflammation-associated monocytes and granulocytes, may be able to actively transport nanoparticles from the blood to the inflamed tissue when they infiltrate the inflamed tissue12, 41, 44. This alternative mechanism was confirmed in the present disclosure in a mouse model of glioma, by showing that highly tumor-infiltrative M-MDSCs can contribute to the tumor accumulation of OH dendrimers. This phenomenon can be leveraged to design M-MDSC-targeting therapeutics for enhanced tumor delivery. [0483] Dendrimer surface functionality affects their interactions with M-MDSC in vivo. Dendrimer surface functionality can significantly affect their in vivo behaviors, such as absorption, distribution, metabolism, elimination (ADME), and toxicity28, 45-47. The effect of dendrimer surface chemistry on cell-level distribution of systemically injected dendrimers was next examined using G6 PAMAM dendrimers with succinamic acid (SA), hydroxyl (OH), and amine (NH2) terminal groups. All three dendrimers showed approximately ~6 nm diameter (number average mean, FIG.10A). When measured in 1×PBS, NH2, OH, and SA dendrimers showed cationic (ζ=32.2±0.5mV), neutral (ζ=5.0±0.2mV), anionic (ζ=-22.4±0.6mV) surface charges (FIG.10B). To trace dendrimers in vivo, all three dendrimers were stably labeled with a minimal amount of Cy5 (5% by wt.%) through previously established conjugation chemistry28 (FIG.10C). All three dendrimers (50mg/kg) were
U1197.70243WO00 146/193 #13587456v2 systematically injected into transgenic mice with established KR158 tumors or age-matched healthy controls. At the dose of 50mg/kg, NH2 dendrimers induced significant toxicities that ultimately led to animal death. The high in vivo toxicity of systemically administrated NH2 dendrimers has been reported previously48. The in vitro toxicity study based on bone marrow-derived M-MDSCs also showed that when the dendrimer dose ≥ 20µg/mL, NH2 dendrimers showed significantly higher toxicity than OH and SA dendrimers (FIG.11A). Therefore, the dose of NH2 dendrimers was lowered to 10mg/kg for the following in vivo studies. [0484] NH2 dendrimers cannot efficiently access M-MDSCs but can be readily taken up by M- MDSCs. Nanoparticles need to efficiently cross the tissue barriers (e.g. the blood vessels and tissue extracellular matrix) before successfully ‘targeting’ the M-MDSCs located in the tissue stroma. How dendrimer surface functionality affects their abilities to ‘target’ M-MDSCs was determined by measuring the percentage of dendrimer-positive M-MDSCs in tissue (% dendrimer+ M-MDSCs). The bone marrow and the tumor are the origin and the destination of M-MDSC recruitment, therefore were selected as the tissues of interest in this study. The % dendrimer+ M-MDSCs was significantly lower for NH2 than other dendrimers in the bone marrow of both KR158 tumor-bearing mice and healthy control (FIG.4A). In the KR158 tumor stroma, NH2 dendrimers also targeted less M-MDSCs (12.3±5.3%) than SA (28±12.7%) and OH dendrimers (19.0±6.3%) (FIG.4B). The lower cell- targeting of NH2 dendrimers was likely due to their lack of ability to cross tissue barriers28. Confocal image in the KR158 tumor showed that NH2 dendrimers were mostly co-localized with the endothelial cells along the blood vessels (FIGs.11B-12A), indicating the NH2 dendrimers with strong cationic surface charge, were not able to move across the blood brain tumor barrier (BBTB) and other tissue barriers28. However, SA and OH dendrimers were able to efficiently cross the BBTB and distribute within the tumor stroma (FIG.12B-12C). Although NH2 dendrimers did not target as many M-MDSCs in the bone marrow, the M-MDSCs that had endocytosed NH2 dendrimers showed similar (for tumor-bearing mice) or higher amounts of intracellular dendrimer (for healthy control) than OH and SA dendrimers (FIG.4C). Specifically, in the bone marrow of healthy mice, the NH2 dendrimer (MFI=1236±453) showed 3.2-fold and 2.7-fold higher M-MDSC-uptake than OH (MFI=384±15) and SA dendrimers (MFI=466±33) (FIG.4D). In the tumor M-MDSCs of KR158 tumor-bearing mice, NH2 dendrimers also showed slightly higher MFI (MFI=685±172) than OH (MFI=442±144) and SA dendrimers (MFI=595±103) (FIG.4E, not statistically significant). Given the dose of NH2 dendrimer is 5-fold lower than OH and SA dendrimers, NH2 dendrimers showed higher capacity for M-MDSC uptake. [0485] M-MDSCs take up SA dendrimers more readily than OH dendrimers. Although the SA and OH dendrimer did not show significant differences in their overall tumor deposition (FIG.13), they did show differential cellular uptake by M-MDSCs. Specifically in the KR158 tumor, M-MDSCs took up more SA dendrimers than OH dendrimers (FIG.4D and E) 24 hours after injection. When
U1197.70243WO00 147/193 #13587456v2 comparing the MFI of dendrimer-positive M-MDSCs in the bone marrow of tumor-bearing mice, the SA (MFI=918±173) was 82% higher than OH (MFI=504±85) (FIG.4D), while for tumor M-MDSCs, the SA (MFI=595±103) was 35% higher than OH (MFI=442±144). In mice that received SA dendrimers, the M-MDSCs compartment and the CX3CR1int compartment (cells that potentially derived from M-MDSCs) accounted for a higher fraction (30.3%) of the dendrimer-positive cells when compared to mice that received OH dendrimers (20.5%) (FIG.4F). The difference between SA and OH dendrimer uptake was further validated in the GL261 tumor model. Within the GL261 tumor stroma, M-MDSCs took up more dendrimers of both SA and OH than other cell subsets (FIG.4G). Tumor M-MDSCs took up 45% higher SA (MFI=2437±250) than OH dendrimers (MFI=1683±245) (FIG.4H), while bone marrow M-MDSCs took up 40% higher SA (MFI=1003±175) than OH dendrimers (MFI=720±264) (FIG.4I). Similar to the KR158 tumor, in the GL261 tumor of SA dendrimer-injected mice, the M-MDSCs compartment and the CX3CR1int compartment accounted for a higher fraction (65.1%) of dendrimer-positive cells than OH dendrimer injected mice (43.2%) (FIG. 4J). In summary, these results showed that SA dendrimers were more efficiently endocytosed by M- MDSCs than OH dendrimers. [0486] Dendrimer-associated serum proteins mediate the interactions between dendrimer and M-MDSCs. Given that M-MDSC showed different capacities for endocytosing NH2, OH, and SA dendrimers in vivo, the mechanism behind the differential uptake was probed by testing the dendrimer uptake in ex vivo generated M-MDSCs. Adapted from an established ex vivo M-MDSC culturing model29, bone marrow cells isolated from transgenic mice were exposed to KR158 conditioned media for 5 days. Flow cytometry analysis showed that the population of CCR2+/CX3CR1+ cells in the bone marrow can be expanded from less than 10% to approximately 59% of the total bone marrow cells ex vivo (FIGs.5A-5B). These cells successfully recapitulate the immune suppressive features and the migration pattern of M-MDSCs in the tumor-bearing mice29. When these ex vivo generated M- MDSCs were exposed to NH2, OH, and SA dendrimers in a serum-free media at a safe dose of 10µg/mL (FIG.11A), a differential uptake of dendrimers was observed. Specifically, the dendrimer uptake was the highest for NH2, followed by SA, and OH dendrimers (FIG.5C). [0487] Although histologic studies have established that the surface charge of dendrimers can significantly affect their interaction with cells46, 47, 49, 50, it is now recognized that for systemically injected nanoparticles, the serum proteins are associated with the nanoparticles are involved in mediating nanoparticle interactions with cells23, 51. To determine how dendrimer-associated serum proteins affect their uptake by M-MDSCs, it was investigated how serum proteins interact with NH2, OH, and SA dendrimers by evaluating the change of ζ-potential after incubating dendrimers with mouse serum (0.86mg/mL). After 30min incubation under 37°C with mouse serum (ζ-potentials=- 11.8±0.37mV), the original ζ-potentials of NH2 (32.2±0.5mV, cationic), OH (5.0±0.2mV, neutral), and SA (-22.4±0.6mV, anionic) all became neutral (NH2: -3.0±1.1mV; OH:0.3±0.6mV; SA:
U1197.70243WO00 148/193 #13587456v2 0.5±0.5mV) (FIG.5D), indicating the surfaces of all three dendrimers were masked by the serum proteins. All dendrimers either with or without serum incubation were then exposed to bone marrow- derived M-MDSCs at an escalating dose of 5µg/mL to 50µg/mL (FIG.5E). Pre-incubating dendrimers with serum significantly decreased the uptake of all dendrimers in a dose-dependent manner (FIG.5F), indicating that ‘naked’ dendrimers and serum protein- ‘coated’ dendrimers interacted with M-MDSCs via different mechanisms. The uptake of NH2 dendrimers had a greater decrease after serum incubation than the OH and SA dendrimers (FIG.5F), indicating the serum protein had a greater influence in mediating the interaction of M-MDSCs with NH2 dendrimers. Serum proteins can be classified into opsonin (enhance uptake) and dysopsonin (reduce uptake)51. To determine the classes of proteins that interreacted with dendrimers of different surface functionalities, dendrimers were incubated with either competent ‘active’ serum or ‘heat-inactivated’ mouse serum under 60°C for 30min (FIG.5G), for NH2 dendrimers, heat-inactivation of serum proteins reduced NH2 dendrimer uptake in a dose-depended manner for up to 60% (FIG.5H), indicating serum proteins associated with NH2 dendrimers actively mediated their uptake by M-MDSCs. However, for both OH and SA dendrimers, heat-inactivation of serum proteins increased their uptake by M-MDSCs up to 24% (OH) and 43% (SA) in a dose-dependent manner (FIG.5H and I), indicating a different class of serum proteins (potentially dysopsonins) might be associated with OH and SA dendrimers. [0488] Historical studies of the ‘protein corona’ associated with nanoparticles are established on nanotherapeutics of 20 – 500nm size ranges with internally encapsulated payloads24, such as lipid nanoparticles (NPs), polymers, and iron oxide NPs. Little is known, however, about NPs with ultra- small architectures, such as dendrimers in the 1 – 20nm size range. Compared to large NPs, dendrimers have sizes similar to proteins and may interact with serum proteins in different stoichiometries and configurations51. Moreover, dendrimers carry payloads on their surfaces. Therefore, the properties of the surface payload will affect how dendrimers interact with M-MDSCs. Here, NH2 and SA dendrimers were used to represent dendrimers carrying drug molecules of acidic and basic properties and compared them with -OH dendrimers (control). The serum proteins associated with SA and OH dendrimers had a similar influence on their interaction with M-MDSCs, while certain serum proteins associated with NH2 dendrimers actively enhance their uptake by M- MDSCs, potentially through receptor-mediated endocytosis. Recently, it has been shown that NH2 dendrimers efficiently interact with IgM and complement protein C352, enhancing their phagocytosis. These ex vivo studies provided a basic mechanistic explanation for the differential in vivo uptake of NH2, OH, and SA dendrimers by M-MDSCs. However, it is not known what specific proteins and their cognate receptors on M-MDSCs mediate the uptake of dendrimers. [0489] M-MDSCs suppress the anti-tumor immune response locally at the TME and globally at the lymphoid organs. To address the systemic immune suppression, nanoparticles need to efficiently target these cells both locally and globally. Using the CCR2RFP/WTCX3CR1GFP/WT transgenic mice that
U1197.70243WO00 149/193 #13587456v2 enable direct surveillance of M-MDSCs, it was shown that that M-MDSCs can infiltrate glioma through peripheral blood in large amounts. Systemically injected hydroxyl dendrimers efficiently target M-MDSCs located in bone marrow, peripheral blood, spleen, and tumor. Within the tumor, M- MDSCs and microglia showed high capacity for endocytosis of hydroxyl dendrimers, and these two cellular compartments accounted for more than half the amount of the hydroxyl dendrimer deposition in the tumor. In the GL261 glioma model, which has a high abundance of infiltrative immune cells, dendrimer showed greater tumor deposition and higher efficiency of M-MDSC-targeting than the KR158 glioma model. The recruitment of M-MDSCs from bone marrow to tumor contributed to the tumor deposition of hydroxyl dendrimers. The surface functionality of dendrimers affects their ability to target M-MDSCs in vivo. Although amine dendrimers had the highest capacity of being endocytosed by M-MDSCs, they could not access these cells as efficiently as hydroxyl or succinamic acid dendrimers, potentially due to the lack of ability to cross tissue barriers. M-MDSCs took up succinamic acid dendrimers more efficiently than hydroxyl dendrimers. Finally, serum proteins can affect how dendrimers interact with M-MDSCs. The serum proteins associated with amine dendrimers significantly enhanced their uptake by M-MDSCs, while serum proteins associated with hydroxyl and succinamic acid dendrimers slightly reduced their uptake by M-MDSCs. Given that dendrimer-based drug conjugates carry drug payload on their surfaces, the payload molecular properties could affect the in vivo fate, such as cell- and tissue-targeting of the final dendrimer-drug conjugates. Example 3: Materials and Methods for Dendrimer-Drug Conjugates for Delivery to M-MDSCs [0490] General Information. The NMR spectra 1H, 1H-decoupled 13C were recorded with TMS as the internal standard. The coupling constants (J values) are given in hertz. The LLL1257 and bi- functional dendrimer58 were synthesized from literature methods. The G6-OH dendrimer was purchased from Dendritech Inc and used after evaporation of their stock solvents. [0491] General procedure for the synthesis of LLL12 derived N-substituted tert-butyl-5- oxopentanoate. To a solution of 5-(tert-butoxy)-5-oxopentanoic acid 2 (0.275 mmol, 1.1 eq, 52 mg) in dichloromethane (3 mL) was added N, N’-dicyclohexylcarbodiimide (0.375 mmol, 1.5 eq, 77 mg) and 4-dimethylaminopyridine (0.05 mmol, 0.2 eq, 6 mg) at 0 oC. To the resulting solution LLL121 (0.25 mmol, 1.0 eq, 76 mg) was added and continued stirring for 10 minutes at the same temperature. Then the reaction mixture was brought to room temperature and continued stirring until completion. After completion of the reaction, the precipitated dicyclohexylurea (DCU) was filtered off and
U1197.70243WO00 150/193 #13587456v2 washed with 5% aqueous acetic acid (5 mL) and water (3 x 5 mL), dried over Na2SO4 and concentrated under reduced pressure. The resulting crude was purified by silica gel column chromatography by gradient elution of EtOAc and hexane (10/90-30/70) to yield 3. [0492] tert-Butyl 5-((5-hydroxy-9,10-dioxo-9,10-dihydroanthracene)-1-sulfonamido)-5- oxopentanoate (3). Yellow solid; Yield 78 mg, 65%; Purified by silica gel column chromatography, Rf = 0.5 (40% EtOAc in hexane); 1H NMR (400 MHz, DMSO-d6) δ 12.10 (s, 1H), 9.43 (s, 1H), 8.72 (d, J = 8.0 Hz, 1H), 8.60 (d, J = 8.0 Hz, 1H), 7.93 (t, J = 8.0 Hz, 1H), 7.69 (d, J = 8.0 Hz, 1H), 7.64 (t, J = 8.0 Hz, 1H), 7.27 (d, J = 8.0 Hz, 1H), 2.38 (t, J = 8.0 Hz, 2H), 2.14 (t, J = 8.0 Hz, 2H), 1.78 (quint, J = 8.0 Hz, 2H), 1.31 (s, 9H); 13C NMR (100 MHz, DMSO-d6) 185.4, 181.2, 171.1, 169.7, 161.2, 138.3, 137.5, 136.5, 134.5, 132.9, 131.5, 123.7, 119.4, 113.9, 79.6, 34.6, 33.0, 28.7, 27.0, 18.5. [0493] General procedure for the synthesis of LLL12 derived 5-oxopentanoic acid. To a solution of 3 (0.165 mmol, 1.0 eq, 78 mg) in dichloromethane (3 mL) was added trifluoroacetic acid (4.125 mmol, 25.0 eq, 470 mg) at room temperature. The resulting reaction mixture was stirred until completion. After completion, the reaction mixture was diluted with dichloromethane (15 mL) and washed with water (3 x 5 mL), dried over Na2SO4 and concentrated under reduced pressure. The resulting crude residue was purified by triturating with hexane to provide 4. [0494] 5-((5-Hydroxy-9,10-dioxo-9,10-dihydroanthracene)-1-sulfonamido)-5-oxopentanoic acid (4). Yellow solid; Yield 66 mg, 96%; Purified by triturating with hexane, Rf = 0.3 (90% EtOAc in hexane); 1H NMR (400 MHz, Acetone-d6) δ 12.05 (s, 1H), 10.65 (s, 1H), 8.64 (d, J = 8.0 Hz, 1H), 8.57 (d, J = 8.0 Hz, 1H), 8.04 (t, J = 8.0 Hz, 1H), 7.72 (t, J = 8.0 Hz, 1H), 7.58 (d, J = 8.0 Hz, 1H), 7.25 (d, J = 8.0 Hz, 1H), 2.43 (t, J = 8.0 Hz, 2H), 2.13 (t, J = 8.0 Hz, 2H), 1.67 (quint, J = 8.0 Hz, 2H); 13C NMR (100 MHz, Acetone-d6) 187.2, 181.4, 173.4, 173.1, 162.0, 139.9, 138.9, 137.6, 135.6, 134.5, 133.7, 132.7, 131.9, 123.8, 119.5, 115.3, 34.9, 32.1, 19.5.
U1197.70243WO00 151/193 #13587456v2 [0495] General procedure for the synthesis of LLL12 derived NHS ester. To a solution of LLL12 derived 5-oxopentanoic acid 4 (0.158 mmol, 1.0 eq, 66 mg) in dry THF (3 mL) was added N, N’- dicyclohexylcarbodiimide (0.174 mmol, 1.1 eq, 36 mg) and N-hydroxysuccinimide 5 (0.174 mmol, 1.1 eq, 20 mg) at 0 oC. The ice bath was removed after 2 h and continued stirring overnight at rt. The white solid DCU was filtered off and solvent was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography by gradient elution of methanol and DCM (05/95-10/90) to yield LLL12 derived NHS ester 6. [0496] 2,5-Dioxopyrrolidin-1-yl 5-((5-hydroxy-9,10-dioxo-9,10-dihydroanthracene)-1- sulfonamido)-5-oxopentanoate (6). Yellow solid; Yield 63 mg, 78%; Purified by silica gel column chromatography, Rf = 0.5 (5% methanol in dichloromethane); 1H NMR (400 MHz, CDCl3) δ 12.13 (s, 1H), 9.39 (s, 1H), 8.74 (d, J = 8.0 Hz, 1H), 8.63 (d, J = 8.0 Hz, 1H), 7.95 (t, J = 8.0 Hz, 1H), 7.71 (d, J = 8.0 Hz, 1H), 7.66 (t, J = 8.0 Hz, 1H), 7.29 (d, J = 8.0 Hz, 1H), 2.73 (s, 4H), 2.55 (t, J = 8.0 Hz, 2H), 2.50 (t, J = 8.0 Hz, 2H), 1.95 (quint, J = 8.0 Hz, 2H). [0497] General procedure for the synthesis of bi-functional dendrimer – Step 1. To a solution of Fmoc-GABA-OH 8 (0.144 mmol, 60 eq, 47 mg) in DMF (3 mL) in a 50 mL round bottom flask under nitrogen atmosphere was added PyBOP (0.216 mmol, 90 eq, 112 mg) in DMF (3 mL) and DIEA (0.288 mmol, 120 eq, 37 mg, 50 uL). The resulting mixture was allowed to stir for an hour in an ice bath. Then, PAMA G6-OH 7 (0.00240 mmol, 1.0 eq, 140 mg) was dissolved in DMF (10 mL) and added to the reaction mixture. The reaction mixture was brought to the rt and continued stirring for 48 h. The solvent was evaporated at 40 oC under reduced pressure. The crude residue was re-dissolved in DMF (2 mL) and dialyzed (membrane cutoff = 12-14 kDa) against DMF for 24 h by changing the DMF every 8 h. The collected solvent was evaporated and subjected to high vacuum for overnight to offer pure off-white semi-solid Fmoc-functionalized bi-functional dendrimer 9. [0498] Fmoc-functionalized bi-functional dendrimer (9). Off-white semi solid; Yield 142 mg, 89%; Purified by dialysis, 1H NMR (400 MHz, DMSO-d6) δ 8.05 (bs, Fmoc aromatic protons), 7.93- 7.97 (m, internal amide NH protons of G6-OH), 7.86 (d, Fmoc aromatic protons), 7.80 (bs, Fmoc aromatic protons), 7.67 (d, Fmoc aromatic protons), 7.39 (t, Fmoc aromatic protons), 7.31 (t, Fmoc aromatic protons), 4.74 (bs, OH protons of G6-OH), 4.29-4.31 (m, Fmoc O-CH2 protons), 4.19-4.21 (m, Fmoc benzylic CH2 proton), 3.45 (bs, N-CH2 protons of G6-OH), 3.25 (bs, CH2 protons of linker), 3.12-3.13 (m, CH2 protons of G6-OH), 3.01-3.02 (d, linker CH2 protons), 2.67 (bs, CH2 protons of G6-OH), 2.45 (bs, CH2 protons of G6-OH), 2.22 (bs, CH2 protons of G6-OH), 1.65 (t, linker CH2 protons).
U1197.70243WO00 152/193 #13587456v2 [0499] General procedure for the synthesis of bi-functional dendrimer – Step 2. The whole batch of above Fmoc-functionalized bi-functional dendrimer was dissolved in DMF (5 mL) and 5 mL of piperidine:DMF (1:4) was added under nitrogen atmosphere. The reaction mixture was stirred for 30 minutes at 0 oC and then solvents were evaporated under vacuum. The crude product was co- evaporated with DMF 5 mL under high vacuum and subjected to dialysis (membrane MW cutoff = 12-14 kDa) for 24 h by changing the DMF after every 8 h. The collected solvent was evaporated and dialyzed agents DI water for 3 h. The collected water was lyophilized to get bi-functional dendrimer 6. [0500] Bi-functional dendrimer (10). White solid; Yield 130 mg, 90%; Purified by dialysis, 1H NMR (400 MHz, DMSO-d6) δ 7.84-8.16 (m, internal amide NH protons of G6-OH), 4.74 (bs, OH protons of G6-OH), 4.02 (d, J = 4.0 Hz, CH2 protons of linker), 4.19-3.37 (bs, N-CH2 protons of G6- OH), 3.07-3.14 (m, CH2 protons of G6-OH and shifted CH2 protons of G6-OH at the site of linker attachment), 2.80 (s, CH2 protons of linker), 2.65 (bs, CH2 protons of G6-OH), 2.43 (bs, CH2 protons of G6-OH), 2.31 (t, J = 4.0 Hz, CH2 protons of linker), 2.21 (bs, CH2 protons of G6-OH), 1.79 (s, linker CH2 protons). [0501] General procedure for the synthesis of dendrimer-drug conjugate via amide linkage. To a solution of bi-functional dendrimer 10 (0.000508 mmol, 1.0 eq, 30 mg) and DIEA (0.0381 mmol, 75.0 eq, 7.0 uL) in DMSO (2 mL) was added LLL12-linker 11 (0.0127 mmol, 25.0 eq, 6.5 mg) in DMSO (1 mL). The resulting mixture was stirred for 48 h at rt. After completion, the reaction mixture was subjected to dialysis against DMF for 24 h by changing the solvent after every 8 h. The obtained solution was evaporated to dryness under reduced pressure at rt, and the final product was dissolved in water and subjected further dialysis against water for 6 h by changing the water after every 2 h. The obtained solution was lyophilized for 32 h to obtain dendrimer-drug conjugate 12 as orange solid (30 mg).
U1197.70243WO00 153/193 #13587456v2 [0502] Dendrimer-drug conjugate via amide linkage (12). Orange solid; Yield 30 mg, 94%; Purified by dialysis; 1H NMR (400 MHz, DMSO-d6) δ 8.54 (bs, LLL12 aromatic CH proton), 8.43 (bs, LLL12 aromatic CH proton), 8.16-7.98 (m, internal amide NH protons of G6-OH), 7.80 (s, LLL12 aromatic CH proton), 7.64 (s, LLL12 aromatic CH proton), 7.33 (s, LLL12 aromatic CH proton), 4.09 (linker CH2 protons), 3.51 (bs, CH2 protons of G6-OH), 3.36 (s, linker CH2 protons), 3.22 (bs, CH2 protons of G6-OH), 2.76 (bs, CH2 protons of G6-OH), 2.57 (s, CH2 protons of G6-OH), 2.34 (s, CH2 protons of G6-OH), 2.11 (s, linker CH2 protons), 1.70 (s, linker CH2 protons). [0503] General procedure for the synthesis of azide linker LLL12 via carbamate linkage. LLL121 (0.2 mmol, 1.10 eq, 60 mg) and azido-PEG-4-nitrophenyl carbonate 13 (0.178 mmol, 1.0 eq, 69 mg) was dissolved in DMF (2 mL) and DMAP (0.356 mmol, 2.0 eq, 43.5 mg) was added to the solution at room temperature. The resulting mixture was stirred for 48 h and diluted with ethyl acetate (15 mL), washed with brine (10 mL), dried over Na2SO4 and concentrated in vacuo. The crude was purified by column chromatography on silica gel by gradient elution of dichloromethane and methanol (95/05 to 85/15) to afford azide linker conjugated LLL1214. [0504] 2-(2-(2-(2-Azidoethoxy)ethoxy)ethoxy)ethyl ((5-hydroxy-9,10-dioxo-9,10- dihydroanthracen-1-yl)sulfonyl)carbamate (14). Yellow solid; Yield 54 mg, 55%; Purified by silica gel column chromatography, Rf = 0.2 (10% methanol in dichloromethane); 1H NMR (400 MHz, methanol-d4) δ 8.62 (d, J = 8.0 Hz, 1H), 8.56 (d, J = 8.0 Hz, 1H), 7.96 (t, J = 8.0 Hz, 1H), 7.66 (overlapped t, J = 8.0 Hz, 1H), 7.65 (overlapped d, J = 8.0 Hz, 1H), 7.21 (d, J = 8.0 Hz, 1H), 4.08 (dt, J = 12.0, 4.0 Hz, 2H), 3.52-3.40 (unresolved m, 12H), 3.22 (dt, J = 12.0, 4.0 Hz, 2H); 13C NMR (100 MHz, methanol-d4) 186.7, 181.0, 161.7, 139.5, 138.5, 137.1, 135.5, 134.2, 133.3, 132.5, 131.8, 123.6, 119.5, 115.0, 70.2 (x 2), 70.1, 70.0, 69.7, 68.4, 65.2, 50.3. [0505] General procedure for the synthesis of propargylated G6-OH dendrimer. To the solution of heptynoic acid 15 (0.0825 mmol, 60 eq, 11 mg) in DMF (1 mL) was added the solution of PyBOP (0.1238 mmol, 90 eq, 64 mg) and DIEA (0.165 mmol, 120 eq, 21 mg, 29 uL) in DMF (2 mL) at 0 oC. After an hour to the resulting solution was added G6-OH dendrimer 7 (0.0014 mmol, 1.0 eq, 80 mg)
U1197.70243WO00 154/193 #13587456v2 in DMF (2 mL). The reaction mixture was brought to the room temperature and stirred for another 48 h. The solvent was evaporated to dryness under high vacuum. The crude was re-dissolved in DMF and dialyzed against DMF for 24 h by at least changing the solvent for 3 times. The collected solvent was evaporated under high vacuum to dryness and the resulting product was dissolved in 2 mL of water, further subjected to pure DI water dialysis for 4 h by changing the solvent two times. The collected water was directly lyophilized to get propargylated dendrimer 16. [0506] Propargylated G6-OH dendrimer (16). White solid; Yield 78 mg, 95%; Purified by dialysis; 1H NMR (400 MHz, DMSO-d6) δ 7.82-8.08 (m, internal amide NH protons of G6-OH), 4.77 (bs, OH protons of G6-OH), 4.01 (t, J = 4.0 Hz, shifted CH2 protons of G6-OH at the site of linker attachment), 3.39-3.45 (m, CH2 protons of G6-OH), 3.28 (t, J = 4.0 Hz, shifted N-CH2 protons of G6- OH at the site of linker attachment), 3.11-3.12 (m, CH2 protons of G6-OH), 2.65 (bs, CH2 protons of G6-OH), 2.51 (t, J = 4.0 Hz, CH protons of linker), 2.44 (bs, CH2 protons of G6-OH), 2.30 (t, J = 8.0 Hz, CH2 protons of linker), 2.21 (bs, CH2 protons of G6-OH), 2.15 (t, J = 8.0 Hz, CH2 protons of linker), 1.56-1.63 (m, CH2 protons of linker), 1.40-1.48 (m, CH2 protons of linker). [0507] General procedure for the synthesis of dendrimer-drug conjugate via carbamate linker. In a 5 mL microwave vial propargylated dendrimer 16 (0.00067 mmol, 1.0 eq, 40 mg) was dissolved in water (750 uL), CuSO4 (0.03 mmol, 1.5 eq, 7.5 mg) in water (125 uL) and sodium ascorbate (0.03 mmol, 1.5 eq, 6 mg) in water (125 uL) were added successively. To this solution drug-linker 14 (0.02 mmol, 30 eq, 11 mg) in THF (1 mL) and DMF (250 uL) were added at rt. The resulting mixture was irradiated in microwave (Biotage microwave reactor) at 45 oC for 7 h. After completion (monitored by TLC), the reaction mixture was diluted with DMF (1 mL) and dialyzed against DMF for 8 h by changing the solvent after every 4 h. The collected solvent was evaporated to dryness and residue was dissolved in water, lyophilized to get the dendrimer-drug conjugate, which was further purified by column chromatography (Sephadex g-25) to get pure dendrimer-drug conjugate 17. [0508] Drug conjugated G6-OH dendrimer via carbamate linker (17). Yellow solid; Yield 40 mg, 88%; Purified by dialysis; 1H NMR (400 MHz, DMSO-d6) δ 8.49 (d, J = 8.0 Hz, drug aromatic CH protons), 8.34 (d, J = 8.0 Hz, drug aromatic CH protons), 8.06 (t, J = 8.0 Hz, drug aromatic CH protons), 7.80-7.94 (m, internal amide NH protons of G6-OH), 7.57 (d, J = 8.0 Hz, drug aromatic CH
U1197.70243WO00 155/193 #13587456v2 protons), 7.32 (d, J = 8.0 Hz, drug aromatic CH protons), 4.68 (bs, OH protons of G6-OH), 4.45 (s, shifted CH2 protons of G6-OH at the site of linker attachment), 3.99 (s, shifted N-CH2 protons of G6- OH at the site of linker attachment), 3.78 (d, J = 4.0 Hz, O-CH2 protons of drug linker), 3.40-3.48 (m, CH2 protons of G6-OH and drug linker CH2 protons are merged) 3.27 (d, J = 4.0 Hz, CH2 protons of linker), 3.10-3.12 (m, CH2 protons of G6-OH), 2.58-2.79 (m, CH2 protons of G6-OH and CH2 protons of propargyl linker), 2.45 (bs, CH2 protons of G6-OH), 2.21-2.32 (m, CH2 protons of G6-OH and CH2 protons of propargyl linker), 1.56 (CH2 protons of propargyl linker). [0509] General procedure for synthesis of LLL12-hydrazone linker (20). Step-1: To a solution of LLL12 (165 mg, 0.54 mmol, 1.0 eq) in ethanol (165 mL) was added acetic acid (1 drop) and hydrazine monohydrate (265uL). The resulting mixture was stirred at 84 oC for 5 h. After completion of the reaction (monitored by TLC), which was stored at -4 oC for 12 h. The crystallized solid was filtered off, filtrate was concentrated to 10% and the precipitated solid was filtered again. Both fractions yielded LLL12-hydrazone 18 in 79% (135 mg). [0510] Step-2: To a solution of LLL12-hydrazone 18 (45 mg, 0.1419 mmol, 1.0 eq) was added acetyl-azide NHS ester 19 (28 mg, 0.1419 mmol, 1.0 eq) and DIEA (74 uL, 55 mg, 0.426 mmol, 3.0 eq) at rt. The resulting reaction mixture was stirred until completion (5 h, monitored by TLC), after completion the reaction mixture was concentrated on high vacuum and purified by silica gel column chromatography (gradient elution of ethyl acetate and hexane, 30/70-50/50) to provide 20. [0511] Synthesis of dendrimer-drug conjugate via hydrazone linker (D-LLL12H) (21). To a solution of bi-functional dendrimer (90 mg, 0.001498 mmol, 1.0 eq) in water (3.2 mL) was added CuSO4.5H2O (17 mg, 0.0674 mmol, 5 mol%) in water (750 uL) and sodium ascorbate (13.4 mg, 0.06742 mmol, 5 mol%) in water (750 uL). To the resulting reaction mixture was added LLL12- hydrazone linker 20 (18 mg, 0.04495 mmol, 30 eq) and DMF (10 mL). The mixture was irradiated in
U1197.70243WO00 156/193 #13587456v2 a microwave (Biotage) for 10 h at 45 oC. After completion (monitored by TLC), the reaction mixture was diluted with DMF (2 mL) and dialyzed (membrane MW cutoff = 12-14 kDa) against DMF for 12 h by changing the solvent every 4 h. The collected solvent was evaporated to dryness and subjected to a high vacuum overnight. The residue was dissolved in water and treated with EDTA (2 mL, 0.5M, pH = 8.0) for 1.5 h, which was then dialyzed against DI water for 12 h by changing the solvent every 3 h. The collected water lyophilized to get the dendrimer-drug conjugate (D-LLL12H); this was further purified by column chromatography (Sephadex G-25) and lyophilized to give pure D-LLL12H conjugate 21. Example 4: Development of Dendrimer-Drug Conjugates for Delivery to M-MDSCs [0512] To overcome the challenges associated with clinical translation of STAT3 inhibitors (because of poor solubility and bioavailability, off-target effects, and limited efficacy in clinical trials), a novel approach based on the dendrimer targeting and uptake studies disclosed herein was developed to systemically deliver LLL12 – a small molecule inhibitor of STAT3 – by formulating LLL12 into dendrimer-based LLL12 conjugates (D-LLL12). This approach is shown in FIG.14. [0513] D-LLL12 was formulated with fast- (D-LLL12C, compound 17) and slow-release (D- LLL12S, compound 12) profiles by reacting LLL12-linkers with bi-functional dendrimers derived from hydroxyl surface dendrimers (G6 PAMAM-OH) over 5 steps. The D-LLL12 conjugates were fully characterized by NMR and HPLC, and IC50 efficacy and toxicity studies were conducted in comparison with LLL12 (FIGs.15-16). [0514] The highest absorption wavelength was identified to in order to determine the minimum detectable concentration of LLL12 by HPLC (FIGs.18A-18E). These absorption wavelengths were used to study the release profiles of the LLL12 from the dendrimer at pH values of 4.5 and 7.4 by HPLC (FIGs.18F-18H). Dose response curves were generated for LLL12 and compound 17, indicating that compound 17 was able to release LLL12, which could then inhibit STAT3 (FIGs.19A- 19F). The toxicity of compound 17 was also shown vs. LLL12, indicating a higher percent of viable cells with compound 17 at higher concentrations of LLL12 (FIG.19G). Table 1. Size and Zeta Potentials of dendrimer (G6-OH) and dendrimer-LLL12 conjugates
U1197.70243WO00 157/193 #13587456v2 Example 5: Efficacy (IC50) Analysis [0515] Cell culture. THP-1STAT3-Luc reporter cell line is cultured in RPMI I640; 10% FBS (Heat- inactivated), 100U/mL penicillin, 100µg/mL streptomycin (1xP/S), 25mM HEPES along with 1.0mM Sodium pyruvate; Puromycin (1µg/mL); 0.05mM 2-mercaptoethanol, puromycin 1µg/mL in the incubator at 37oC in 5% CO2. [0516] Experimental procedure. A black clear bottom 96-well plate assay is used for IC50 using the suspension THP-1STAT3-Luc cells (FIG.35). [0517] Day 0 – Cells are seeded at 60,000 cells/well in 100µl of media (RPMI 1640 - without phenol red, 25mM HEPES, 100U/mL penicillin, 100µg/mL streptomycin (1xP/S). The plate is incubated for 4 hours. The free drug LLL12 and its dendrimer drug conjugate formulations - D-LLL12C (17) (CS0177) and D-LLL12S (12) (CS0186) are added to the wells at concentrations ranging from 11.1µg/mL ^ 3.7µg/mL ^ 1.23µg/mL ^ 0.4111µg/mL ^ 0.137µg/mL ^ 0.0456µg/mL ensuring triplicates per concentration. The plate is further incubated for 72 hours treatment period. Controls (set up in triplicates) – 1. Cells only (Background control); 2. Cells + 15ul (150µg/mL) D-Luc (Endogenous STAT3 expression); 3. Cells + 15ul (150µg/mL) D-Luc + 10µg/mL IL6 (Maximum activated STAT3 expression). [0518] Day 4 – For the activation of STAT3, 10µg/mL IL6 is added to all the cells treated with the drug and the control wells. The plate is returned to the incubator at 37oC in 5% CO2 for 5 hours.15µl of 150µg/ml D-Luc is added to all the wells and appropriate controls and the plate is incubated for 10- 15 minutes. The luminescence is recorded by the SpectraMax iD3 plate reader. Background luminescence is subtracted from the experimental and the control wells. The concentration of test compounds versus bioluminescence is plotted and the data is analyzed by GraphPad prism. [0519] Results. The IC50 study demonstrates that free drug (LLL12) has highest potency with the lowest IC50 value of 0.305 µg/mL (FIG.36A). The dendrimer drug conjugates (21, 17, and 12) show lower potency (IC50=1.07µg/mL for 21; IC50=34.79µg/mL for 17; IC50=6.47µg/mL for 12). The IC50 study was repeated in three independent experiments (FIG.36B). The average IC50 and the standard deviation for each formulation are shown in Table 2. Table 2. IC50 values for LLL12 and dendrimer-LLL12 conjugates.
U1197.70243WO00 158/193 #13587456v2 Example 6: Toxicity Study [0520] Cell culture. THP-1 cells are cultured in RPMI I640; 10% FBS (Heat-inactivated), 100U/mL penicillin, 100µg/mL streptomycin (1xP/S), 25mM HEPES; 0.05mM 2-mercaptoethanol in the incubator at 37oC in 5% CO2. [0521] Experimental procedure. A black clear bottom 96-well plate assay is used with the suspension THP-1 cells. [0522] Day 0 – Cells are seeded at 60,000 cells/well in 100µl of media (RPMI 1640 - without phenol red, 25mM HEPES, 100U/mL penicillin, 100µg/mL streptomycin (1xP/S). The plate is incubated for 4 hours. The free drug LLL12 and its dendrimer drug conjugate formulations - 17 (CS0177) and 12 (CS0186) are added to the wells at concentrations ranging from 300µg/mL ^ 0.0456µg/mL ensuring triplicates per concentration. The plate is further incubated for 72 hours treatment period. Controls (set up in triplicates) – 1. No cells + CellTitre blue reagent (Background control); 2. Untreated cells (Negative control); 3. Cells + 1% Triton X-100 (Positive control). [0523] Day 4 – 1% Triton X-100 is added as a positive control to account for maximum cell death. The plate is incubated for 3 hours at 37oC in 5% CO2. Add 20ul/well CellTitre-Blue Reagent. Incubate further for 3 hours. The fluorescence is recorded by the SpectraMax iD3 plate reader at 560/590nm. The fluorescence values of the culture medium background are subtracted from all the experimental and control wells. The concentration of test compounds versus % viability is plotted, and the data is analyzed by GraphPad prism. [0524] Results. The dendrimer-drug conjugates (17 and 12) exhibit lower cytotoxicity compared to the free drug, especially at the concentrations range of 0.4 - 4 µg/mL (FIGs.37A-37C). Example 7: Therapeutic Window [0525] The therapeutic window measures the dose range of a drug that provides safe and effective therapy with minimal adverse effects. Therapeutic window is the dose ranging from the onset of efficacy to the dose at which only 50% cells are viable (50% viability). By overlaying the IC50 curve (curve with open circles) and the viability curve (curve with shaded triangles) in the same plot, the therapeutic window (shaded region) of each formulation can be assessed (FIGs.38A-38E). [0526] Free LLL12 showed a very narrow therapeutic window (0.38 compared to dendrimer-drug conjugates.12 showed the widest therapeutic window (2.31), followed by 17 (1.68), and 21 (1.21). [0527] References [0528] 1. Kumar M, Kulkarni P, Liu S, Chemuturi N, Shah DK. Nanoparticle biodistribution coefficients: A quantitative approach for understanding the tissue distribution of nanoparticles. Adv Drug Deliv Rev.2023;194:114708. Epub 20230120. doi: 10.1016/j.addr.2023.114708. PubMed PMID: 36682420.
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U1197.70243WO00 165/193 #13587456v2 EQUIVALENTS AND SCOPE [0592] In the articles such as “a,” “an,” and “the” may mean one or more than one unless indicated to the contrary or otherwise evident from the context. Embodiments or descriptions that include “or” between one or more members of a group are considered satisfied if one, more than one, or all of the group members are present in, employed in, or otherwise relevant to a given product or process unless indicated to the contrary or otherwise evident from the context. The invention includes embodiments in which exactly one member of the group is present in, employed in, or otherwise relevant to a given product or process. The invention includes embodiments in which more than one, or all of the group members are present in, employed in, or otherwise relevant to a given product or process. [0593] Furthermore, the disclosure encompasses all variations, combinations, and permutations in which one or more limitations, elements, clauses, and descriptive terms from one or more of the listed claims is introduced into another claim. For example, any claim that is dependent on another claim can be modified to include one or more limitations found in any other claims that is dependent on the same base claim. Where elements are presented as lists, e.g., in Markush group format, each subgroup of the elements is also disclosed, and any element(s) can be removed from the group. It should it be understood that, in general, where the invention, or aspects of the invention, is/are referred to as comprising particular elements and/or features, certain embodiments of the disclosure or aspects of the disclosure consist, or consist essentially of, such elements and/or features. For purposes of simplicity, those embodiments have not been specifically set forth in haec verba herein. It is also noted that the terms “comprising” and “containing” are intended to be open and permits the inclusion of additional elements or steps. Where ranges are given, endpoints are included. Furthermore, unless otherwise indicated or otherwise evident from the context and understanding of one of ordinary skill in the art, values that are expressed as ranges can assume any specific value or sub–range within the stated ranges in different embodiments of the invention, to the tenth of the unit of the lower limit of the range, unless the context clearly dictates otherwise. [0594] This application refers to various issued patents, published patent applications, journal articles, and other publications, all of which are incorporated herein by reference. If there is a conflict between any of the incorporated references and the instant specification, the specification shall control. In addition, any particular embodiment of the present invention that falls within the prior art may be explicitly excluded from any one or more of the embodiments. Because such embodiments are deemed to be known to one of ordinary skill in the art, they may be excluded even if the exclusion is not set forth explicitly herein. Any particular embodiment of the invention can be excluded from any embodiment, for any reason, whether or not related to the existence of prior art. [0595] Those skilled in the art will recognize or be able to ascertain using no more than routine experimentation many equivalents to the specific embodiments described herein. The scope of the present embodiments described herein is not intended to be limited to the above Description, but rather is as set forth in the appended embodiments. Those of ordinary skill in the art will appreciate
U1197.70243WO00 166/193 #13587456v2 that various changes and modifications to this description may be made without departing from the spirit or scope of the present invention, as defined in the following claims.
U1197.70243WO00 167/193 #13587456v2

Claims

CLAIMS What is claimed is: 1. A compound of Formula (I): or a pharmaceutically acceptable salt thereof, wherein: Zn is an nth generation polyamidoamine (PAMAM) dendrimer; n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; each instance of Y is independently –ORA, –N(RA)2, or –C(=O)ORA; each instance of RA is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; each instance of L is independently optionally substituted alkylene, optionally substituted heterocyclylene, optionally substituted arylene, optionally substituted heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted alkylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene are independently replaced with –O–, –NR1–, –S–, –S(=O)–, –S(=O)2–, –C(=O)–, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene; each instance of R1 is independently hydrogen or optionally substituted alkyl; each instance of X is independently a conjugated agent; p is an integer between 1 and 2(n+2), inclusive; q is an integer between 0 and 2(n+2)-1, inclusive; and the sum of p and q is 2(n+2). 2. A compound of Formula (I′): or a pharmaceutically acceptable salt thereof, wherein: Zn is an nth generation polyamidoamine (PAMAM) dendrimer; n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; each instance of Y is independently –ORA, –N(RA)2, or –C(=O)ORA; each instance of RA is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally
U1197.70243WO00 168/193 #13587456v2 substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; each instance of L is independently optionally substituted alkylene, optionally substituted heterocyclylene, optionally substituted arylene, optionally substituted heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted alkylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene are independently replaced with –O–, –NR1–, –S–, –S(=O)–, –S(=O)2–, –C(=O)–, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene; each instance of R1 is independently hydrogen or optionally substituted alkyl; each instance of X is independently selected from formulae (ii), (iii), (iv), (v), (vi), (vii), (viii), (ix), (x), and (xi):
U1197.70243WO00 169/193 #13587456v2 p is an integer between 1 and 2(n+2), inclusive; q is an integer between 0 and 2(n+2)-1, inclusive; and the sum of p and q is 2(n+2). 3. A compound of Formula (I′): or a pharmaceutically acceptable salt thereof, wherein: Zn is an nth generation polyamidoamine (PAMAM) dendrimer; n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; each instance of Y is independently –ORA, –N(RA)2, or –C(=O)ORA; each instance of RA is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; each instance of L is independently optionally substituted alkylene, optionally substituted heterocyclylene, optionally substituted arylene, optionally substituted heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted alkylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene are independently replaced with –O–, –NR1–, –S–, –S(=O)–, –S(=O)2–, –C(=O)–, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene; each instance of R1 is independently hydrogen or optionally substituted alkyl; each instance of X is independently selected from formulae (ii), (iii), (iv), (v), (vi), (vii), (viii), (ix), and (x):
U1197.70243WO00 170/193 #13587456v2 p is an integer between 1 and 2(n+2), inclusive; q is an integer between 0 and 2(n+2)-1, inclusive; and the sum of p and q is 2(n+2). 4. The compound of any one of claims 1-3, wherein the compound is of Formula (I-a): or a pharmaceutically acceptable salt thereof.
U1197.70243WO00 171/193 #13587456v2
5. The compound of any one of claims 1-4, or a pharmaceutically acceptable salt thereof, wherein the compound of Formula (I) comprises between about 2% and about 15% by weight of X. 6. The compound of any one of claims 1-5, or a pharmaceutically acceptable salt thereof, wherein the compound of Formula (I) comprises between about 5% and about 10% by weight of X. 7. The compound of any one of claims 1 or 4-6, or a pharmaceutically acceptable salt thereof, wherein the conjugated agent is a radical of an agent. 8. The compound of claim 7, or a pharmaceutically acceptable salt thereof, wherein the agent is a hydrophobic agent, an anionic agent, or a cationic agent. 9. The compound of any one of claims 7 or 8, or a pharmaceutically acceptable salt thereof, wherein the agent is a STAT3 inhibitor, a TLR7 agonist, an IDO inhibitor, an NOS inhibitor, an arginase inhibitor, a PD-L1 inhibitor, a protein kinase B inhibitor, a CD73 inhibitor, or a STING agonist. 10. The compound of any one of claims 1-9, or a pharmaceutically acceptable salt thereof, wherein each instance of X is selected from formulae (ii), (iii), and (iv). 11. The compound of any one of claims 1-9, or a pharmaceutically acceptable salt thereof, wherein each instance of X is selected from formulae (v), (vi), and (vii). 12. The compound of any one of claims 1-9, or a pharmaceutically acceptable salt thereof, wherein each instance of X is selected from formulae (viii), (ix), and (x). 13. The compound of any one of claims 1-12, wherein the compound is selected from Formulae (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), and (XI):
U1197.70243WO00 172/193 #13587456v2 U1197.70243WO00 173/193 #13587456v2 or a pharmaceutically acceptable salt thereof: 14. A compound of Formula (II): or a pharmaceutically acceptable salt thereof, wherein: Zn is an nth generation polyamidoamine (PAMAM) dendrimer; n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; each instance of Y is independently –ORA, –N(RA)2, or –C(=O)ORA; each instance of RA is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; each instance of L is independently optionally substituted alkylene, optionally substituted heterocyclylene, optionally substituted arylene, optionally substituted heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted alkylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene are independently replaced with –O–, –NR1–, –S–, –S(=O)–, –S(=O)2–, –C(=O)–, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene; each instance of R1 is independently hydrogen or optionally substituted alkyl; p is an integer between 1 and 2(n+2), inclusive; q is an integer between 0 and 2(n+2)-1, inclusive; and the sum of p and q is 2(n+2).
U1197.70243WO00 174/193 #13587456v2
15. A compound of Formul or a pharmaceutically acceptable salt thereof, wherein: Zn is an nth generation polyamidoamine (PAMAM) dendrimer; n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; each instance of Y is independently –ORA, –N(RA)2, or –C(=O)ORA; each instance of RA is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; each instance of L is independently optionally substituted alkylene, optionally substituted heterocyclylene, optionally substituted arylene, optionally substituted heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted alkylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene are independently replaced with –O–, –NR1–, –S–, –S(=O)–, –S(=O)2–, –C(=O)–, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene; each instance of R1 is independently hydrogen or optionally substituted alkyl; p is an integer between 1 and 2(n+2), inclusive; q is an integer between 0 and 2(n+2)-1, inclusive; and the sum of p and q is 2(n+2). 16. The compound of any one of claims 1-15, or a pharmaceutically acceptable salt thereof, wherein each instance of Y is independently –ORA. 17. The compound of any one of claims 1-16, or a pharmaceutically acceptable salt thereof, wherein each instance of RA is independently hydrogen, optionally substituted alkyl, optionally substituted alkynyl, or optionally substituted heteroalkyl. 18. The compound of any one of claims 1-17, or a pharmaceutically acceptable salt thereof,
U1197.70243WO00 175/193 #13587456v2 wherein at least one instance of RA is hydrogen. 19. The compound of any one of claims 1-18, or a pharmaceutically acceptable salt thereof, wherein at least one instance of RA is optionally substituted alkyl, optionally substituted alkynyl, or optionally substituted heteroalkyl. 20. The compound of any one of claims 1-19, or a pharmaceutically acceptable salt thereof, wherein at least one instance of RA is of formula: (i), wherein RA1 is optionally substituted alkyl, optionally substituted alkynyl, or optionally substituted heteroalkyl. 21. The compound of any one of claims 1-20, or a pharmaceutically acceptable salt thereof, wherein at least one instance of RA is of formula: (i), wherein RA1 is optionally substituted C1-10 alkyl, optionally substituted C1-10 alkynyl, or optionally substituted C1-10 heteroalkyl. 22. The compound of any one of claims 1-21, or a pharmaceutically acceptable salt thereof, 23. The compound of any one of claims 1-22, wherein the compound is of Formula (II-a): or a pharmaceutically acceptable salt thereof. 24. The compound of any one of claims 1-23, wherein the compound is of Formula (II-b):
U1197.70243WO00 176/193 #13587456v2 or a pharmaceutically acceptable salt thereof. 25. The compound of any one of claims 1-22, wherein the compound is of Formula (XI-a): or a pharmaceutically acceptable salt thereof. 26. The compound of any one of claims 1-22 or 25, wherein the compound is of Formula (XI-b): or a pharmaceutically acceptable salt thereof. 27. The compound of any one of claims 1-26, or a pharmaceutically acceptable salt thereof, wherein n is 4, 5, 6, 7, 8, 9, or 10. 28. The compound of any one of claims 1-27, or a pharmaceutically acceptable salt thereof, wherein n is 6. 29. The compound of any one of claims 1-28, or a pharmaceutically acceptable salt thereof, wherein at least one instance of L is optionally substituted alkylene, optionally substituted
U1197.70243WO00 177/193 #13587456v2 heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted alkylene or optionally substituted heteroarylene are independently replaced with –O–, –NR1–, –S–, –S(=O)–, –S(=O)2–, –C(=O)–, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene. 30. The compound of any one of claims 1-29, or a pharmaceutically acceptable salt thereof, wherein at least one instance of L is optionally substituted alkylene, optionally substituted heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted alkylene or optionally substituted heteroarylene are independently replaced with –O–, –NR1–, –C(=O)–, or optionally substituted heteroarylene. 31. The compound of any one of claims 1-30, or a pharmaceutically acceptable salt thereof, wherein at least one instance of L is optionally substituted C1-20 alkylene, optionally substituted 5-6 membered heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted C1-20 alkylene or optionally substituted 5-6 membered heteroarylene are independently replaced with –O–, –NR1–, –C(=O)–, or optionally substituted heteroarylene. 32. The compound of any one of claims 1-31, or a pharmaceutically acceptable salt thereof, wherein at least one instance of L is optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene containing 1, 2, or 3 ring N atoms, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted linear C1-20 alkylene or optionally substituted 5-6 membered heteroarylene containing 1, 2, or 3 ring N atoms are independently replaced with –O–, –NR1–, –C(=O)–, or optionally substituted heteroarylene. 33. The compound of any one of claims 1-32, or a pharmaceutically acceptable salt thereof, wherein at least one instance of L comprises optionally substituted alkylene, optionally substituted heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted alkylene or optionally substituted heteroarylene are independently replaced with –O–, –NR1–, –S–, –S(=O)–, –S(=O)2–, –C(=O)–, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene. 34. The compound of any one of claims 1-33, or a pharmaceutically acceptable salt thereof, wherein at least one instance of L comprises optionally substituted alkylene, optionally substituted heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in
U1197.70243WO00 178/193 #13587456v2 the optionally substituted alkylene or optionally substituted heteroarylene are independently replaced with –O–, –NR1–, –C(=O)–, or optionally substituted heteroarylene. 35. The compound of any one of claims 1-34, or a pharmaceutically acceptable salt thereof, wherein at least one instance of L comprises optionally substituted C1-20 alkylene, optionally substituted 5-6 membered heteroarylene, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted C1-20 alkylene or optionally substituted 5-6 membered heteroarylene are independently replaced with –O–, –NR1–, –C(=O)–, or optionally substituted heteroarylene. 36. The compound of any one of claims 1-35, or a pharmaceutically acceptable salt thereof, wherein at least one instance of L comprises optionally substituted linear C1-20 alkylene, optionally substituted 5-6 membered heteroarylene containing 1, 2, or 3 ring N atoms, or a combination thereof, optionally wherein one or more backbone carbon atoms in the optionally substituted linear C1-20 alkylene or optionally substituted 5-6 membered heteroarylene containing 1, 2, or 3 ring N atoms are independently replaced with –O–, –NR1–, –C(=O)–, or optionally substituted heteroarylene. 37. The compound of any one of claims 1-36, or a pharmaceutically acceptable salt thereof, wherein at least one instance of L comprises optionally substituted C1-20 alkylene, wherein one or more backbone carbon atoms in the optionally substituted C1-20 alkylene are independently replaced with –O–, –NR1–, –C(=O)–, or optionally substituted heteroarylene. 38. The compound of any one of claims 1-37, or a pharmaceutically acceptable salt thereof, wherein at least one instance of L comprises optionally substituted linear C1-20 alkylene, wherein one or more backbone carbon atoms in the optionally substituted linear C1-20 alkylene are independently replaced with –O–, –NR1–, –C(=O)–, or optionally substituted heteroarylene. 39. The compound of any one of claims 1-38, or a pharmaceutically acceptable salt thereof, , ,
U1197.70243WO00 179/193 #13587456v2 , wherein each x is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; and each y is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. 40. The compound of any one of claims 1-39, or a pharmaceutically acceptable salt thereof, 41. The compound of any one of claims 1-40, or a pharmaceutically acceptable salt thereof, wherein at least one instance of L comprises optionally substituted heteroarylene. 42. The compound of any one of claims 1-41, or a pharmaceutically acceptable salt thereof, wherein at least one instance of L comprises optionally substituted 5-6 membered heteroarylene. 43. The compound of any one of claims 1-42, or a pharmaceutically acceptable salt thereof, wherein at least one instance of L comprises optionally substituted 5-6 membered heteroarylene having 1, 2, or 3 ring heteroatoms selected from O, N, and S. 44. The compound of any one of claims 1-43, or a pharmaceutically acceptable salt thereof, wherein at least one instance of L comprises optionally substituted 5-6 membered heteroarylene having 3 ring N atoms. 45. The compound of any one of claims 1-44, or a pharmaceutically acceptable salt thereof,
U1197.70243WO00 180/193 #13587456v2 wherein at least one instance of L comprises 46. The compound of any one of claims 1-45, or a pharmaceutically acceptable salt thereof, wherein at least one instance of L comprises . 47. The compound of any one of claims 1-46, or a pharmaceutically acceptable salt thereof, 48. The compound of any one of claims 1-47, or a pharmaceutically acceptable salt thereof, wherein the sum of p and q is 256. 49. The compound of any one of claims 1-48, or a pharmaceutically acceptable salt thereof, wherein p is an integer between 1 and 40, inclusive, and q is an integer between 216 and 255, inclusive. 50. The compound of any one of claims 1-49, or a pharmaceutically acceptable salt thereof, wherein p is an integer between 10 and 30, inclusive, and q is an integer between 226 and 246, inclusive. 51. The compound of any one of claims 1-50, or a pharmaceutically acceptable salt thereof, wherein p is an integer between 17 and 23, inclusive, and q is an integer between 233 and 239, inclusive; or p is an integer between 12 and 18, inclusive, and q is an integer between 238 and 244, inclusive. 52. The compound of any one of claims 1-51, or a pharmaceutically acceptable salt thereof, U1197.70243WO00 181/193 #13587456v2 wherein p is 20 and q is 236; or p is 15 and q is 241. 53. The compound of any one of claims 14, 16-24, or 27-52, or a pharmaceutically acceptable salt thereof, wherein the compound of Formula (II) comprises between about 2% and about 15% by 54. The compound of any one of claims 14, 16-24, or 27-53, or a pharmaceutically acceptable salt thereof, wherein the compound of Formula (II) comprises between about 5% and about 10% by mass 55. The compound of any one of claims 1-14, 16-24, or 27-54, wherein the compound is of 56. The compound of any one of claims 1-14, 16-24, or 27-55, wherein the compound is of
U1197.70243WO00 182/193 #13587456v2 Formula (II-c-i) or Formula (II-d-i): 57. The compound of any one of claims 1-14, 16-24, or 27-56, wherein the compound is of formula: , or a pharmaceutically acceptable salt thereof. 58. The compound of any one of claims 15-22 or 25-52, or a pharmaceutically acceptable salt thereof, wherein the compound of Formula (XI) comprises between about 2% and about 15% by mass
U1197.70243WO00 183/193 #13587456v2 59. The compound of any one of claims 15-22, 25-52, or 58, or a pharmaceutically acceptable salt thereof, wherein the compound of Formula (XI) comprises between about 5% and about 10% by mass 60. The compound of any one of claims 1, 2, 4-13, 15-22, 25-52, 58, or 59, wherein the compound is of or a pharmaceutically acceptable salt thereof. 61. The compound of any one of claims 1, 2, 4-13, 15-22, 25-52, or 58-60, wherein the
U1197.70243WO00 184/193 #13587456v2 compound is of or a pharmaceutically acceptable salt thereof. 62. The compound of any one of claims 1, 2, 4-13, 15-22, 25-52, or 58-61, wherein the compound is of formula: , or a pharmaceutically acceptable salt thereof. 63. A pharmaceutical composition comprising a compound any one of claims 1-62, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient. 64. A method of treating or preventing a disease or disorder in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of the compound of any one of claims 1-62, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 63. 65. The method of claim 64, wherein the disease or disorder is a proliferative disease. 66. The method of claim 65, wherein the proliferative disease is cancer. 67. The method of claim 66, wherein the cancer comprises a solid tumor.
U1197.70243WO00 185/193 #13587456v2
68. The method of any one of claims 66 or 67, wherein the cancer is brain tumor (e.g., glioma, glioblastoma), pancreatic cancer, or ovarian cancer. 69. A method of modulating a target protein, the method comprising administering to a subject in need thereof or contacting a cell, tissue, or biological sample with an effective amount of the compound of any one of claims 1-62, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 63. 70. The method of claim 69, wherein modulating the target protein comprises inhibition of the target protein. 71. The method of claim 69, wherein modulating the target protein comprises agonism of the target protein. 72. The method of any one of claims 69-71, wherein the target protein is signal transducer and activator of transcription 3 (STAT3), Toll-like receptor 7 (TLR7), indoleamine 2,3-dioxygenase (IDO), nitric oxide synthase (NOS), arginase, programmed death-ligand 1 (PD-L1), protein kinase B, cluster of differentiation 73 (CD73), or stimulator of interferon genes (STING). 73. The method of any one of claims 69-72, wherein the cell is a myeloid derived suppressor cell (MDSC). 74. The method of any one of claims 69-72, wherein the cell is a tumor-infiltrating monocyte (TIM). 75. The method of any one of claims 64-74, wherein the administration is by injection. 76. A method of preparing a compound of Formula (II-c):
U1197.70243WO00 186/193 #13587456v2 or a pharmaceutically acceptable salt thereof, the method comprising: providing a reaction vessel; adding to the reaction vessel a compound of Formula (XIII-a): (XIII-a), or a salt thereof; adding to the reaction vessel a compound of Formula (XIV-a): or a salt thereof; and obtaining the compound of Formula (II-c), or pharmaceutically acceptable salt thereof; wherein: Zn is an nth generation polyamidoamine (PAMAM) dendrimer; n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; each instance of RA is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; each of p and p1 is independently an integer between 1 and 2(n+2), inclusive; each of q and q1 is independently an integer between 0 and 2(n+2)-1, inclusive; the sum of p and q is 2(n+2); the sum of p1 and q1 is 2(n+2); p is less than or equal to p1; and q is greater than or equal to q1.
U1197.70243WO00 187/193 #13587456v2
77. A method of preparing a compound of Formula (II-d): or a pharmaceutically acceptable salt thereof, the method comprising: providing a reaction vessel; adding to the reaction vessel a compound of Formula (XIII-b): or a salt thereof; adding to the reaction vessel a compound of Formula (XIV-b): or a salt thereof; and obtaining the compound of Formula (II-d), or pharmaceutically acceptable salt thereof; wherein: Zn is an nth generation polyamidoamine (PAMAM) dendrimer; n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; each instance of RA is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; each of p and p2 is independently an integer between 1 and 2(n+2), inclusive; each of q and q2 is independently an integer between 0 and 2(n+2)-1, inclusive; the sum of p and q is 2(n+2); the sum of p2 and q2 is 2(n+2); p is less than or equal to p2; and q is greater than or equal to q2.
U1197.70243WO00 188/193 #13587456v2
78. A method of preparing a compound of Formula (XI-d): or a pharmaceutically acceptable salt thereof, the method comprising: providing a reaction vessel; adding to the reaction vessel a compound of Formula (XIII-b): or a salt thereof; adding to the reaction vessel a compound of Formula (XIV-c): or a salt thereof; and obtaining the compound of Formula (XI-d), or pharmaceutically acceptable salt thereof; wherein: Zn is an nth generation polyamidoamine (PAMAM) dendrimer; n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; each instance of RA is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; each of p and p2 is independently an integer between 1 and 2(n+2), inclusive; each of q and q2 is independently an integer between 0 and 2(n+2)-1, inclusive; the sum of p and q is 2(n+2); the sum of p2 and q2 is 2(n+2);
U1197.70243WO00 189/193 #13587456v2 p is less than or equal to p2; and q is greater than or equal to q2. 79. A compound made by the method of any one of claims 76-78. 80. A compound, or a pharmaceutically acceptable salt thereof, made by the method comprising: providing a reaction vessel; adding to the reaction vessel a compound of Formula (III-a): or a salt thereof; adding to the reaction vessel a compound of Formula (IV-a): or a salt thereof; and obtaining the compound, or pharmaceutically acceptable salt thereof; wherein: Zn is an nth generation polyamidoamine (PAMAM) dendrimer; n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; each instance of RA is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; p1 is an integer between 1 and 2(n+2), inclusive; q1 is an integer between 0 and 2(n+2)-1, inclusive; and the sum of p1 and q1 is 2(n+2). 81. A method of preparing a compound, or a pharmaceutically acceptable salt thereof, the method comprising: providing a reaction vessel; adding to the reaction vessel a compound of Formula (III-b):
U1197.70243WO00 190/193 #13587456v2 or a salt thereof; adding to the reaction vessel a compound of Formula (IV-b): or a salt thereof; and obtaining the compound, or pharmaceutically acceptable salt thereof; wherein: Zn is an nth generation polyamidoamine (PAMAM) dendrimer; n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; each instance of RA is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; p2 is an integer between 1 and 2(n+2), inclusive; q2 is an integer between 0 and 2(n+2)-1, inclusive; and the sum of p2 and q2 is 2(n+2). 82. A method of preparing a compound, or a pharmaceutically acceptable salt thereof, the method comprising: providing a reaction vessel; adding to the reaction vessel a compound of Formula (XIII-b): or a salt thereof; adding to the reaction vessel a compound of Formula (XIV-c):
U1197.70243WO00 191/193 #13587456v2 or a salt thereof; and obtaining the compound, or pharmaceutically acceptable salt thereof; wherein: Zn is an nth generation polyamidoamine (PAMAM) dendrimer; n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; each instance of RA is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; p2 is an integer between 1 and 2(n+2), inclusive; q2 is an integer between 0 and 2(n+2)-1, inclusive; and the sum of p2 and q2 is 2(n+2). 83. A kit comprising: the compound of any one of claims 1-62, or the pharmaceutical composition of claim 63; and instructions for its use.
U1197.70243WO00 192/193 #13587456v2
PCT/US2025/014062 2024-01-31 2025-01-31 Selective delivery of agents to myeloid-derived suppressor cells with a dendrimer-based formulation Pending WO2025166181A1 (en)

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Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
BHASIN ET AL.: "Antiproliferative activities and SAR studies of substituted anthraquinones and 1,4-naphthoquinones", BIOORGANIC & MEDICINAL CHEMISTRY LETTERS, vol. 23, 2013, pages 6864 - 6867, XP028788000, DOI: 10.1016/j.bmcl.2013.09.098 *
CHANDRASEKAR DURAIRAJ, SISTLA RAMAKRISHNA, AHMAD FARHAN J., KHAR ROOP K., DIWAN PRAKASH V.: "Folate coupled poly(ethyleneglycol) conjugates of anionic poly(amidoamine) dendrimer for inflammatory tissue specific drug delivery", JOURNAL OF BIOMEDICAL MATERIALS RESEARCH PART A, JOHN WILEY & SONS, US, vol. 82A, no. 1, 1 July 2007 (2007-07-01), US , pages 92 - 103, XP093345622, ISSN: 1549-3296, DOI: 10.1002/jbm.a.31122 *
DATABASE PUBCHEM COMPOUND 13 September 2005 (2005-09-13), XP093345388, Database accession no. 4140276 *
NEGAR TAGHAVI POURIANAZAR: "Bioapplications of poly(amidoamine) (PAMAM) dendrimers in nanomedicine", JOURNAL OF NANOPARTICLE RESEARCH, SPRINGER NETHERLANDS, DORDRECHT, vol. 16, no. 4, 1 January 2014 (2014-01-01), Dordrecht, pages 2342 - 2342-38, XP009195124, ISSN: 1388-0764, DOI: 10.1007/s11051-014-2342-1 *

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