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WO2025128639A1 - Hydrogels comprenant des nanoparticules stabilisées par des protéines pour une libération prolongée - Google Patents

Hydrogels comprenant des nanoparticules stabilisées par des protéines pour une libération prolongée Download PDF

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WO2025128639A1
WO2025128639A1 PCT/US2024/059479 US2024059479W WO2025128639A1 WO 2025128639 A1 WO2025128639 A1 WO 2025128639A1 US 2024059479 W US2024059479 W US 2024059479W WO 2025128639 A1 WO2025128639 A1 WO 2025128639A1
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composition
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therapeutic agent
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Daniel S. Kohane
Wonmin Choi
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Boston Childrens Hospital
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Boston Childrens Hospital
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K17/00—Carrier-bound or immobilised peptides; Preparation thereof
    • C07K17/02—Peptides being immobilised on, or in, an organic carrier
    • C07K17/08—Peptides being immobilised on, or in, an organic carrier the carrier being a synthetic polymer
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/445—Non condensed piperidines, e.g. piperocaine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • A61K9/0024—Solid, semi-solid or solidifying implants, which are implanted or injected in body tissue
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/141—Intimate drug-carrier mixtures characterised by the carrier, e.g. ordered mixtures, adsorbates, solid solutions, eutectica, co-dried, co-solubilised, co-kneaded, co-milled, co-ground products, co-precipitates, co-evaporates, co-extrudates, co-melts; Drug nanoparticles with adsorbed surface modifiers
    • A61K9/146—Intimate drug-carrier mixtures characterised by the carrier, e.g. ordered mixtures, adsorbates, solid solutions, eutectica, co-dried, co-solubilised, co-kneaded, co-milled, co-ground products, co-precipitates, co-evaporates, co-extrudates, co-melts; Drug nanoparticles with adsorbed surface modifiers with organic macromolecular compounds

Definitions

  • a composition comprising a hydrogel and a protein- stabilized particle, wherein: the hydrogel comprises a crosslinked protein matrix; the protein-stabilized particle comprises a therapeutic agent or a diagnostic agent; and the protein-stabilized particle is embedded in the crosslinked protein matrix.
  • methods of treating a disease or condition comprising administering an effective amount of the composition.
  • kits comprising the composition and instructions for using the composition.
  • 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.
  • HPLC high pressure liquid chromatography
  • C1–6 is intended to encompass C1, C2, C3, C4, C5, C6, C1–6, C1–5, C 1–4 , C 1–3 , C 1–2 , C 2–6 , C 2–5 , C 2–4 , C 2–3 , C 3–6 , C 3–5 , C 3–4 , C 4–6 , C 4–5 , and C 5–6 .
  • C 1-6 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.
  • 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”).
  • branched alkyl refers to a radical of a branched saturated hydrocarbon group having from 1 to 20 carbon atoms (“branched C1–20 alkyl”), for example, isopropyl, t-butyl, sec-butyl, iso-butyl, neopentyl, isopentyl, and neoheptyl.
  • unbranched alkyl is the same as a straight-chain or linear alkyl group, i.e., an alkyl group having no alkyl branching groups. In some embodiments, an alkyl group has 1 to 12 carbon atoms (“C1–12 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 (“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 (“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 (“C1–4 alkyl”).
  • an alkyl group has 1 to 3 carbon atoms (“C1–3 alkyl”). In some embodiments, an alkyl group has 1 to 2 carbon atoms (“C1–2 alkyl”). In some #13447332v2 embodiments, an alkyl group has 1 carbon atom (“C1 alkyl”). In some embodiments, 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 (C5) (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 (C7), 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 C 1–12 alkyl (such as unsubstituted C 1–6 alkyl, e.g., ⁇ CH 3 (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 C 1–12 alkyl such as unsubstituted C 1–6 alkyl, e.g.
  • the alkyl group is a substituted C 1–12 alkyl (such as substituted C1–6 alkyl, e.g., –CH2F, –CHF2, –CF3, –CH2CH2F, –CH2CHF2, –CH2CF3, or benzyl (Bn)).
  • 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”).
  • an alkenyl group has 1 to 12 carbon atoms (“C 1–12 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 (“C 1–9 alkenyl”). In some embodiments, an alkenyl group has 1 to 8 carbon atoms (“C 1–8 alkenyl”). In some embodiments, an alkenyl group has 1 to 7 carbon atoms (“C1–7 alkenyl”).
  • an alkenyl group has 1 to 6 carbon atoms (“C1–6 alkenyl”). In some embodiments, an alkenyl group has 1 to 5 carbon atoms (“C 1–5 alkenyl”). In some embodiments, an alkenyl group has 1 to 4 carbon atoms (“C 1–4 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 (“C 1 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 (C 3 ), 1-butenyl (C 4 ), 2-butenyl (C 4 ), butadienyl (C 4 ), and the like.
  • Examples of C 1–6 alkenyl groups include the aforementioned C 2-4 alkenyl groups as well as pentenyl (C 5 ), pentadienyl (C5), hexenyl (C6), and the like.
  • C 1-20 alkynyl refers to a radical of a straight-chain, unbranched 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”).
  • linear alkynyl refers to a radical of a straight-chain, unbranched 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”).
  • an alkynyl group has 1 to 10 carbon atoms (“C 1-10 alkynyl”).
  • an alkynyl group has 1 to 9 carbon atoms (“C 1-9 alkynyl”).
  • an alkynyl group has 1 to 8 carbon atoms (“C1-8 alkynyl”). In some embodiments, an alkynyl group has 1 to 7 carbon atoms (“C 1-7 alkynyl”). In some embodiments, an alkynyl group has 1 to 6 carbon atoms (“C 1-6 alkynyl”). In some embodiments, an alkynyl group has 1 to 5 carbon atoms (“C 1-5 alkynyl”). In some embodiments, an alkynyl group has 1 to 4 carbon atoms (“C1-4 alkynyl”). In some embodiments, an alkynyl group has 1 to 3 carbon atoms (“C1-3 alkynyl”).
  • an alkynyl group has 1 to 2 carbon atoms (“C 1-2 alkynyl”). In some embodiments, an alkynyl group has 2 to 10 carbon atoms (“C 2–10 alkynyl”). In some embodiments, an alkynyl group has 2 to 9 carbon atoms (“C2–9 alkynyl”). In some embodiments, an alkynyl group has 2 to 8 carbon atoms (“C 2–8 alkynyl”). In some embodiments, an alkynyl group has 2 to 7 carbon atoms (“C 2–7 alkynyl”). In some embodiments, an alkynyl group has 2 to 6 carbon atoms (“C 2–6 alkynyl”).
  • an alkynyl group has 2 to 5 carbon atoms (“C2–5 alkynyl”). In some embodiments, an alkynyl group has 2 to 4 carbon atoms (“C 2–4 alkynyl”). In some embodiments, an alkynyl group has 2 to 3 carbon atoms (“C 2–3 alkynyl”). In some embodiments, an alkynyl group has 2 carbon atoms (“C2 alkynyl”). In some embodiments, an alkynyl group has 1 carbon atom (“C1 alkynyl”). The one or more carbon-carbon triple bonds can be internal (such as in 2- butynyl) or terminal (such as in 1-butynyl).
  • C 1-4 alkynyl groups include, without limitation, methylidynyl (C 1 ), ethynyl (C 2 ), 1-propynyl (C 3 ), 2-propynyl (C 3 ), 1-butynyl (C 4 ), 2- butynyl (C4), and the like.
  • C1-6 alkenyl groups include the aforementioned C2-4 alkynyl groups as well as pentynyl (C 5 ), hexynyl (C 6 ), and the like.
  • C 2–4 alkynyl groups include, without limitation, ethynyl (C 2 ), 1–propynyl (C 3 ), 2–propynyl (C 3 ), 1–butynyl (C4), 2–butynyl (C4), and the like.
  • Examples of C2–6 alkenyl groups include the aforementioned C 2–4 alkynyl groups as well as pentynyl (C 5 ), hexynyl (C 6 ), and the like. Additional examples of alkynyl include heptynyl (C 7 ), octynyl (C 8 ), and the like.
  • each #13447332v2 instance of an alkynyl group is independently unsubstituted (an “unsubstituted alkynyl”) or substituted (a “substituted alkynyl”) with one or more substituents.
  • the alkynyl group is an unsubstituted C 1-20 alkynyl.
  • the alkynyl group is a substituted C1-20 alkynyl.
  • the alkynyl group is an optionally substituted C2-20 alkynyl.
  • carbocyclyl 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.
  • a carbocyclyl group has 3 to 14 ring carbon atoms (“C 3-14 carbocyclyl”).
  • a carbocyclyl group has 3 to 13 ring carbon atoms (“C3-13 carbocyclyl”).
  • a carbocyclyl group has 3 to 12 ring carbon atoms (“C3-12 carbocyclyl”).
  • a carbocyclyl group has 3 to 11 ring carbon atoms (“C 3-11 carbocyclyl”). In some embodiments, a carbocyclyl group has 3 to 10 ring carbon atoms (“C 3-10 carbocyclyl”). In some embodiments, a carbocyclyl group has 3 to 8 ring carbon atoms (“C3-8 carbocyclyl”). In some embodiments, a carbocyclyl group has 3 to 7 ring carbon atoms (“C3-7 carbocyclyl”). In some embodiments, a carbocyclyl group has 3 to 6 ring carbon atoms (“C 3-6 carbocyclyl”).
  • a carbocyclyl group has 4 to 6 ring carbon atoms (“C4-6 carbocyclyl”). In some embodiments, a carbocyclyl group has 5 to 6 ring carbon atoms (“C5-6 carbocyclyl”). In some embodiments, a carbocyclyl group has 5 to 10 ring carbon atoms (“C 5-10 carbocyclyl”).
  • Exemplary C 3-6 carbocyclyl groups include cyclopropyl (C3), cyclopropenyl (C3), cyclobutyl (C4), cyclobutenyl (C4), cyclopentyl (C5), cyclopentenyl (C5), cyclohexyl (C6), cyclohexenyl (C6), cyclohexadienyl (C6), and the like.
  • Exemplary C3-8 carbocyclyl groups include the aforementioned C 3-6 carbocyclyl groups as well as cycloheptyl (C 7 ), cycloheptenyl (C 7 ), cycloheptadienyl (C 7 ), cycloheptatrienyl (C 7 ), cyclooctyl (C 8 ), cyclooctenyl (C8), bicyclo[2.2.1]heptanyl (C7), bicyclo[2.2.2]octanyl (C8), and the like.
  • Exemplary C3-10 carbocyclyl groups include the aforementioned C3-8 carbocyclyl groups as well as cyclononyl (C 9 ), cyclononenyl (C 9 ), cyclodecyl (C 10 ), cyclodecenyl (C 10 ), octahydro-1H- indenyl (C9), decahydronaphthalenyl (C10), spiro[4.5]decanyl (C10), and the like.
  • Exemplary C3-8 carbocyclyl groups include the aforementioned C3-10 carbocyclyl groups as well as cycloundecyl (C 11 ), spiro[5.5]undecanyl (C 11 ), cyclododecyl (C 12 ), cyclododecenyl (C 12 ), cyclotridecane (C 13 ), cyclotetradecane (C14), and the like.
  • the carbocyclyl group is either monocyclic (“monocyclic carbocyclyl”) or polycyclic (e.g., containing a fused, bridged or spiro ring system such as a bicyclic system (“bicyclic carbocyclyl”) or tricyclic system (“tricyclic carbocyclyl”)) and can be saturated or can contain one or more carbon-carbon double or triple bonds.
  • the carbocyclyl group is a substituted C 3-14 carbocyclyl.
  • “carbocyclyl” is a monocyclic, saturated carbocyclyl group having from 3 to 14 ring carbon atoms (“C3-14 cycloalkyl”).
  • a cycloalkyl group has 3 to 10 ring carbon atoms (“C 3-10 cycloalkyl”).
  • a cycloalkyl group has 3 to 8 ring carbon atoms (“C3-8 cycloalkyl”).
  • a cycloalkyl group has 3 to 6 ring carbon atoms (“C3-6 cycloalkyl”).
  • a cycloalkyl group has 4 to 6 ring carbon atoms (“C 4-6 cycloalkyl”). In some embodiments, a cycloalkyl group has 5 to 6 ring carbon atoms (“C 5-6 cycloalkyl”). In some embodiments, a cycloalkyl group has 5 to 10 ring carbon atoms (“C5-10 cycloalkyl”). Examples of C5-6 cycloalkyl groups include cyclopentyl (C5) and cyclohexyl (C5). Examples of C3-6 cycloalkyl groups include the aforementioned C 5-6 cycloalkyl groups as well as cyclopropyl (C 3 ) and cyclobutyl (C 4 ).
  • a heterocyclyl group is a 5–10 membered non-aromatic ring system having ring carbon atoms and 1–4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5–10 membered heterocyclyl”).
  • a heterocyclyl group is a 5–8 membered non-aromatic ring system having ring carbon atoms and 1–4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5–8 membered heterocyclyl”).
  • Exemplary 3-membered heterocyclyl groups containing 1 heteroatom include azirdinyl, oxiranyl, and thiiranyl.
  • Exemplary 4-membered heterocyclyl groups containing 1 heteroatom include azetidinyl, oxetanyl, and thietanyl.
  • Exemplary 5-membered heterocyclyl groups containing 1 heteroatom include tetrahydrofuranyl, dihydrofuranyl, tetrahydrothiophenyl, dihydrothiophenyl, pyrrolidinyl, dihydropyrrolyl, and pyrrolyl-2,5-dione.
  • Exemplary 6-membered heterocyclyl groups containing 3 heteroatoms include triazinyl.
  • Exemplary 7-membered heterocyclyl groups containing 1 #13447332v2 heteroatom include azepanyl, oxepanyl, and thiepanyl.
  • Exemplary 8-membered heterocyclyl groups containing 1 heteroatom include azocanyl, oxecanyl and thiocanyl.
  • aryl refers to a radical of a monocyclic or polycyclic (e.g., bicyclic or tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10, or 14 pi electrons shared in a cyclic array) having 6–14 ring carbon atoms and zero heteroatoms provided in the aromatic ring system (“C 6-14 aryl”).
  • an aryl group has 6 ring carbon atoms (“C 6 aryl”; e.g., phenyl).
  • an aryl group has 10 ring carbon atoms (“C10 aryl”; e.g., naphthyl such as 1–naphthyl and 2-naphthyl).
  • an aryl group has 14 ring carbon atoms (“C 14 aryl”; e.g., anthracyl).
  • Aryl also includes ring systems wherein the aryl ring, as defined above, is fused with one or more carbocyclyl or heterocyclyl groups wherein the radical or point of attachment is on the aryl ring, and in such instances, the number of carbon atoms continue to designate the number of carbon atoms in the aryl ring system.
  • each instance of an aryl group is independently unsubstituted (an “unsubstituted aryl”) or substituted (a “substituted aryl”) with one or more substituents.
  • 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”).
  • the point of attachment can be a carbon or nitrogen atom, as valency permits.
  • Heteroaryl polycyclic ring systems can include one or more heteroatoms in one or both rings.
  • Polycyclic heteroaryl groups wherein one ring does not contain a heteroatom e.g., indolyl, quinolinyl, carbazolyl, and the like
  • the point of attachment can be on either ring, e.g., either the ring bearing a heteroatom (e.g., 2-indolyl) or the ring that does not contain a heteroatom (e.g., 5-indolyl).
  • the heteroaryl is substituted or unsubstituted, 5- or 6-membered, monocyclic heteroaryl, wherein 1, 2, 3, or 4 atoms in the heteroaryl ring system are independently oxygen, nitrogen, or sulfur.
  • the heteroaryl is substituted or unsubstituted, 9- or 10-membered, bicyclic heteroaryl, wherein 1, 2, 3, or 4 atoms in the heteroaryl ring system are independently oxygen, nitrogen, or sulfur.
  • a heteroaryl group is a 5-10 membered aromatic ring system 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 (“5-10 membered heteroaryl”).
  • a heteroaryl group is a 5-8 membered aromatic ring system 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 (“5-8 membered heteroaryl”).
  • a heteroaryl group is a 5-6 membered aromatic ring system 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 (“5-6 membered heteroaryl”).
  • the 5-6 membered heteroaryl has 1–3 ring heteroatoms selected from nitrogen, oxygen, and sulfur.
  • the 5-6 membered heteroaryl has 1–2 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5-6 membered heteroaryl has 1 ring heteroatom selected from nitrogen, oxygen, and sulfur. Unless otherwise specified, each instance of a heteroaryl group is independently unsubstituted (an “unsubstituted heteroaryl”) or substituted (a “substituted heteroaryl”) with one or more substituents. In certain embodiments, the heteroaryl group is an unsubstituted 5-14 membered heteroaryl. In certain embodiments, the heteroaryl group is a substituted 5-14 membered heteroaryl.
  • Exemplary 5-membered heteroaryl groups containing 1 heteroatom include pyrrolyl, furanyl, and thiophenyl.
  • Exemplary 5-membered heteroaryl groups containing 2 heteroatoms #13447332v2 include imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl.
  • Exemplary 5- membered heteroaryl groups containing 3 heteroatoms include triazolyl, oxadiazolyl, and thiadiazolyl.
  • Exemplary 5-membered heteroaryl groups containing 4 heteroatoms include tetrazolyl.
  • Exemplary 6-membered heteroaryl groups containing 1 heteroatom include pyridinyl.
  • Exemplary 5,6- bicyclic heteroaryl groups include indolyl, isoindolyl, indazolyl, benzotriazolyl, benzothiophenyl, isobenzothiophenyl, benzofuranyl, benzoisofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzoxadiazolyl, benzthiazolyl, benzisothiazolyl, benzthiadiazolyl, indolizinyl, and purinyl.
  • Exemplary 6,6-bicyclic heteroaryl groups include naphthyridinyl, pteridinyl, quinolinyl, isoquinolinyl, cinnolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl.
  • Exemplary tricyclic heteroaryl groups include phenanthridinyl, dibenzofuranyl, carbazolyl, acridinyl, phenothiazinyl, phenoxazinyl, and phenazinyl.
  • Heteroaralkyl is a subset of “alkyl” and refers to an alkyl group substituted by a heteroaryl group, wherein the point of attachment is on the alkyl moiety.
  • the term “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.
  • 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.
  • 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.
  • the disclosure is not limited in any manner by the exemplary substituents described herein.
  • each carbon atom substituent is independently halogen, substituted (e.g., substituted with one or more halogen) or unsubstituted C1-6 alkyl, ⁇ OR aa , ⁇ SR aa , ⁇ N(R bb )2, –CN, –SCN, or –NO2.
  • each carbon atom substituent is independently halogen, substituted (e.g., substituted with one or more halogen moieties) or unsubstituted C 1–10 alkyl, ⁇ OR aa , ⁇ SR aa , ⁇ N(R bb )2, –CN, –SCN, or –NO2, wherein R aa is hydrogen, substituted (e.g., substituted with one or more halogen) or unsubstituted C1–10 alkyl, an oxygen protecting group (e.g., silyl, TBDPS, TBDMS, TIPS, TES, TMS, MOM, THP, t-Bu, Bn, allyl, acetyl, pivaloyl, or benzoyl) when attached to an oxygen atom, or a sulfur protecting group (e.g., acetamidomethyl, t-Bu, 3-nitro-2-pyridine sulfenyl, 2-pyridine-s
  • the molecular weight of a carbon atom substituent is lower than 250, lower than 200, lower than 150, lower than 100, or lower than 50 g/mol.
  • a carbon atom substituent consists of carbon, hydrogen, fluorine, chlorine, bromine, iodine, oxygen, sulfur, nitrogen, and/or silicon atoms.
  • a carbon atom substituent consists of carbon, hydrogen, fluorine, chlorine, bromine, iodine, oxygen, sulfur, and/or nitrogen atoms.
  • a carbon atom substituent consists of carbon, hydrogen, fluorine, chlorine, bromine, and/or iodine atoms.
  • thiol refers to the group –SH.
  • amino refers to the group ⁇ NH2.
  • substituted amino by extension, refers to a monosubstituted amino, a disubstituted amino, or a trisubstituted amino. In certain embodiments, the “substituted amino” is a monosubstituted amino or a disubstituted amino group.
  • trisubstituted amino refers to an amino group wherein the nitrogen atom directly attached to the parent molecule is substituted with three groups, and includes groups selected from ⁇ N(R bb ) 3 and ⁇ N(R bb ) 3 + X ⁇ , wherein R bb and X ⁇ are as defined herein.
  • acyl groups include aldehydes ( ⁇ CHO), carboxylic acids ( ⁇ CO 2 H), ketones, acyl halides, esters, amides, imines, carbonates, carbamates, and ureas.
  • Acyl substituents include, but are not limited to, any of the substituents described herein, that result in the formation of a stable moiety (e.g., aliphatic, alkyl, alkenyl, alkynyl, heteroaliphatic, heterocyclic, aryl, heteroaryl, acyl, oxo, imino, thiooxo, cyano, isocyano, amino, azido, nitro, hydroxyl, thiol, halo, aliphaticamino, heteroaliphaticamino, alkylamino, heteroalkylamino, arylamino, heteroarylamino, alkylaryl, arylalkyl, aliphaticoxy, heteroaliphaticoxy, alkyl
  • sil refers to the group –Si(R aa )3, wherein R aa is as defined herein.
  • Nitrogen atoms can be substituted or unsubstituted as valency permits, and include primary, secondary, tertiary, and quaternary nitrogen atoms.
  • Nitrogen protecting groups are well known in the art and include those described in detail in Protecting Groups in Organic #13447332v2 Synthesis, T. W. Greene and P. G. M. Wuts, 3 rd edition, John Wiley & Sons, 1999, incorporated herein by reference.
  • each nitrogen protecting group, together with the nitrogen atom to which the nitrogen protecting group is attached is independently selected from the group consisting of methyl carbamate, ethyl carbamate, 9- fluorenylmethyl carbamate (Fmoc), 9-(2-sulfo)fluorenylmethyl carbamate, 9-(2,7- dibromo)fluoroenylmethyl carbamate, 2,7-di-t-butyl-[9-(10,10-dioxo-10,10,10,10- tetrahydrothioxanthyl)]methyl carbamate (DBD-Tmoc), 4-methoxyphenacyl carbamate (Phenoc), 2,2,2-trichloroe
  • each nitrogen protecting group is independently selected from the group consisting of p-toluenesulfonamide (Ts), benzenesulfonamide, 2,3,6-trimethyl-4-methoxybenzenesulfonamide (Mtr), 2,4,6- trimethoxybenzenesulfonamide (Mtb), 2,6-dimethyl-4-methoxybenzenesulfonamide (Pme), 2,3,5,6-tetramethyl-4-methoxybenzenesulfonamide (Mte), 4-methoxybenzenesulfonamide (Mbs), 2,4,6-trimethylbenzenesulfonamide (Mts), 2,6-dimethoxy-4-methylbenzenesulfonamide (iMds), 2,2,5,7,8-pentamethylchroman-6-sulfonamide (Pmc), methanesulfonamide (Ms),
  • Ts p-toluenesulfonamide
  • Mtr
  • each nitrogen protecting group is independently selected from the group #13447332v2 consisting of phenothiazinyl-(10)-acyl derivatives, N’-p-toluenesulfonylaminoacyl derivatives, N’-phenylaminothioacyl derivatives, N-benzoylphenylalanyl derivatives, N-acetylmethionine derivatives, 4,5-diphenyl-3-oxazolin-2-one, N-phthalimide, N-dithiasuccinimide (Dts), N-2,3- diphenylmaleimide, N-2,5-dimethylpyrrole, N-1,1,4,4-tetramethyldisilylazacyclopentane adduct (STABASE), 5-substituted 1,3-dimethyl-1,3,5-triazacyclohexan-2-one, 5-substituted 1,3
  • two instances of a nitrogen protecting group together with the nitrogen atoms to which the nitrogen protecting groups are attached are N,N’-isopropylidenediamine.
  • at least one nitrogen protecting group is Bn, Boc, Cbz, Fmoc, trifluoroacetyl, triphenylmethyl, acetyl, or Ts.
  • 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”).
  • each oxygen protecting group is selected from the group consisting of methyl, methoxymethyl (MOM), methylthiomethyl (MTM), t-butylthiomethyl, (phenyldimethylsilyl)methoxymethyl (SMOM), benzyloxymethyl (BOM), p- methoxybenzyloxymethyl (PMBM), (4-methoxyphenoxy)methyl (p-AOM), guaiacolmethyl (GUM), t-butoxymethyl, 4-pentenyloxymethyl (POM), siloxymethyl, 2-methoxyethoxymethyl (MEM), 2,2,2-trichloroethoxymethyl, bis(2-chloroethoxy)methyl, 2- (trimethylsilyl)ethoxymethyl (SEMOR), tetrahydropyranyl (THP), 3-bro
  • At least one oxygen protecting group is silyl, TBDPS, TBDMS, TIPS, TES, TMS, MOM, THP, t-Bu, Bn, allyl, acetyl, pivaloyl, or benzoyl.
  • each sulfur atom substituent is independently substituted (e.g., substituted with one or more halogen) or unsubstituted C1-6 alkyl or a sulfur protecting group.
  • the substituent present on a sulfur atom is a sulfur protecting group (also referred to as a “thiol protecting group”).
  • the molecular weight of a substituent is lower than 250, lower than 200, lower than 150, lower than 100, or lower than 50 g/mol.
  • a substituent consists of carbon, hydrogen, fluorine, chlorine, bromine, iodine, oxygen, sulfur, nitrogen, and/or silicon atoms.
  • a substituent consists of carbon, hydrogen, fluorine, chlorine, bromine, iodine, oxygen, sulfur, and/or nitrogen atoms. In certain embodiments, a substituent consists of carbon, hydrogen, fluorine, chlorine, bromine, and/or iodine atoms. In certain embodiments, a substituent consists of carbon, hydrogen, fluorine, and/or chlorine atoms. In certain embodiments, a substituent comprises 0, 1, 2, or 3 hydrogen bond donors. In certain embodiments, a substituent comprises 0, 1, 2, or 3 hydrogen bond acceptors.
  • 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 – , HCO 3 ⁇ , HSO 4 – , 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
  • Exemplary counterions which may be multivalent include CO3 2 ⁇ , HPO4 2 ⁇ , PO 3 ⁇ 4 , B4O7 2 ⁇ , SO4 2 ⁇ , S2O3 2 ⁇ , #13447332v2 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, fum
  • non-hydrogen group refers to any group that is defined for a particular variable that is not hydrogen.
  • 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.
  • 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,
  • 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.
  • solvate refers to forms of the compound, or a salt thereof, that are associated with a solvent, usually by a solvolysis reaction. This physical association may include hydrogen bonding.
  • solvents include water, methanol, ethanol, acetic acid, DMSO, THF, diethyl ether, and the like.
  • the compounds described herein may be prepared, e.g., in crystalline form, and may be solvated. Suitable solvates include pharmaceutically acceptable solvates and further include both stoichiometric solvates and non-stoichiometric solvates.
  • the solvate will be capable of isolation, for example, when one or more solvent molecules are incorporated in the crystal lattice of a crystalline solid.
  • “Solvate” encompasses both solution-phase and isolatable solvates.
  • Representative solvates include hydrates, ethanolates, and methanolates.
  • the term “hydrate” refers to a compound that is associated with water. Typically, the number of the water molecules contained in a hydrate of a compound is in a definite ratio to the number of the compound molecules in the hydrate. Therefore, a hydrate of a compound may be represented, for example, by the general formula R ⁇ x H2O, wherein R is the compound, and x is a number greater than 0.
  • a given compound may form more than one type of hydrate, including, e.g., monohydrates (x is 1), lower hydrates (x is a number greater than 0 and smaller than 1, e.g., hemihydrates (R ⁇ 0.5 H 2 O)), and polyhydrates (x is a number greater than 1, e.g., dihydrates (R ⁇ 2 H 2 O) and hexahydrates (R ⁇ 6 H 2 O)).
  • tautomers or “tautomeric” refers to two or more interconvertible compounds resulting from at least one formal migration of a hydrogen atom and at least one change in valency (e.g., a single bond to a double bond, a triple bond to a single bond, or vice versa).
  • Tautomerizations i.e., the reaction providing a tautomeric pair
  • exemplary tautomerizations include keto-to-enol, amide-to-imide, lactam-to-lactim, enamine-to- imine, and enamine-to-(a different enamine) tautomerizations.
  • isomers 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”.
  • stereoisomers that differ in the arrangement of their atoms in space are termed “stereoisomers”.
  • 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”.
  • 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 #13447332v2 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”.
  • the term “crystalline” or “crystalline form” refers to a solid form substantially exhibiting three-dimensional order.
  • a crystalline form of a solid is a solid form that is substantially not amorphous.
  • the X-ray powder diffraction (XRPD) pattern of a crystalline form includes one or more sharply defined peaks.
  • the term “co-crystal” refers to a crystalline structure comprising at least two different components (e.g., a compound disclosed herein and an acid), wherein each of the components is independently an atom, ion, or molecule. In certain embodiments, none of the components is a solvent. In certain embodiments, at least one of the components is a solvent. A co-crystal of a compound disclosed herein and an acid is different from a salt formed from a compound disclosed herein and the acid.
  • a compound disclosed herein is complexed with the acid in a way that proton transfer (e.g., a complete proton transfer) from the acid to a compound disclosed herein easily occurs at room temperature.
  • a compound disclosed herein is complexed with the acid in a way that proton transfer from the acid to a compound disclosed herein does not easily occur at room temperature.
  • Co-crystals may be useful to improve the properties (e.g., solubility, stability, and ease of formulation) of a compound disclosed herein.
  • polymorph refers to a crystalline form of a compound (or a salt, hydrate, or solvate thereof). All polymorphs have the same elemental composition. Different crystalline forms usually have different X-ray diffraction patterns, infrared spectra, melting points, density, hardness, crystal shape, optical and electrical properties, stability, and solubility. Recrystallization solvent, rate of crystallization, storage temperature, and other factors may cause one crystal form to dominate.
  • Various polymorphs of a compound can be prepared by crystallization under different conditions.
  • small molecule refers to molecules, whether naturally occurring or artificially created (e.g., via chemical synthesis) that have a relatively low molecular weight.
  • a small molecule is an organic compound (e.g., it contains carbon).
  • the small molecule may contain multiple carbon-carbon bonds, stereocenters, and other functional groups (e.g., amines, hydroxyl, carbonyls, and heterocyclic rings, etc.).
  • the molecular weight of a small molecule is not more than about 1,000 g/mol, not more than about 900 g/mol, not more than about 800 g/mol, not more than about 700 g/mol, not more than about 600 g/mol, not #13447332v2 more than about 500 g/mol, not more than about 400 g/mol, not more than about 300 g/mol, not more than about 200 g/mol, or not more than about 100 g/mol.
  • the molecular weight of a small molecule is at least about 100 g/mol, at least about 200 g/mol, at least about 300 g/mol, at least about 400 g/mol, at least about 500 g/mol, at least about 600 g/mol, at least about 700 g/mol, at least about 800 g/mol, or at least about 900 g/mol, or at least about 1,000 g/mol. Combinations of the above ranges (e.g., at least about 200 g/mol and not more than about 500 g/mol) are also possible.
  • the small molecule is a therapeutically active agent such as a drug (e.g., a molecule approved by the U.S.
  • the small molecule may also be complexed with one or more metal atoms and/or metal ions.
  • the small molecule is also referred to as a “small organometallic molecule.”
  • Preferred small molecules are biologically active in that they produce a biological effect in animals, preferably mammals, more preferably humans. Small molecules include, but are not limited to, radionuclides and imaging agents.
  • the small molecule is a drug.
  • the drug is one that has already been deemed safe and effective for use in humans or animals by the appropriate governmental agency or regulatory body. For example, drugs approved for human use are listed by the FDA under 21 C.F.R.
  • a “subject” to which administration is contemplated includes, but is not limited to, humans (i.e., a male or female of any age group, e.g., a pediatric subject (e.g., infant, child, adolescent) or adult subject (e.g., young adult, middle–aged adult, or senior adult)) and/or other non–human animals, for example, mammals (e.g., primates (e.g., cynomolgus monkeys, rhesus monkeys); commercially relevant mammals such as cattle, pigs, horses, sheep, goats, cats, and/or dogs) and birds (e.g., commercially relevant birds such as chickens, ducks, geese, and/or turkeys).
  • mammals e.g., primates (e.g., cynomolgus monkeys, rhesus monkeys); commercially relevant mammals such as cattle, pigs, horses, sheep, goats, cats, and/or dogs) and birds (
  • the animal is a mammal.
  • the animal may be a male or female and at any stage of development.
  • a non–human animal may be a transgenic animal.
  • the term “patient” refers to a human subject in need of treatment of a disease or disorder.
  • tissue 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, organelles (such as obtained by lysing cells and separating the components thereof by centrifugation or #13447332v2 otherwise).
  • 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
  • cell fractions, fragments, organelles such as obtained by lysing cells and separating the components thereof by centrifugation or #13447332v2 otherwise.
  • biological samples include blood, serum, urine, semen, fecal matter, cerebrospinal fluid, interstitial fluid, mucus, 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.
  • Biological samples also include those biological samples that are transgenic, such as a transgenic oocyte, sperm cell, blastocyst, embryo, fetus, donor cell, or cell nucleus, or cells or cell lines derived from biological samples.
  • 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 disclosure is delivered.
  • a tissue may be an abnormal or unhealthy tissue, which may need to be treated.
  • a 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 tissue is the central nervous system.
  • the tissue is the brain.
  • administer refers to implanting, absorbing, ingesting, injecting, inhaling, or otherwise introducing a compound, or a pharmaceutical composition thereof.
  • treatment refers to reversing, alleviating, delaying the onset of, or inhibiting the progress of a “pathological condition” (e.g., a disease, disorder, or condition, or one or more signs or symptoms thereof) described herein.
  • pathological condition e.g., a disease, disorder, or condition, or one or more signs or symptoms thereof
  • treatment may be administered after one or more signs or symptoms have developed or have been observed.
  • treatment may be administered in the absence of signs or symptoms of the disease or condition.
  • the subject is at a higher risk of developing the disease or disorder or at a higher risk of regression of the disease or disorder than an average healthy member of a population of subjects.
  • condition e.g., chronic obstructive pulmonary disease
  • disease e.g., chronic pulmonary disease
  • disorder e.g., pulmonary disease.
  • an effective amount of a compound described herein refers to an amount sufficient to elicit the desired biological response, i.e., treating the condition.
  • the effective amount of a compound described herein may vary depending on such factors as the desired biological endpoint, the pharmacokinetics of the compound, the condition being treated, the mode of administration, and the age and health of the subject.
  • An effective amount encompasses therapeutic and prophylactic treatment.
  • an effective amount of a compound may reduce the tumor burden or stop the growth or spread of a tumor.
  • 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 or causes of the condition, or enhances the therapeutic efficacy of another therapeutic agent.
  • a therapeutically effective amount is an amount sufficient, e.g.
  • a therapeutically effective amount is an amount sufficient for treating a disease or condition.
  • a “prophylactically effective amount” of a compound described herein is an amount sufficient to prevent a condition, or one or more signs or 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.
  • prophylactically effective amount can encompass an amount that improves overall prophylaxis or enhances the prophylactic efficacy of another prophylactic agent.
  • therapeutic compound therapeutic agent
  • therapeutic moiety refer to any substance having therapeutic properties that produce a desired, usually beneficial, effect.
  • therapeutic compounds, agents, and moieties may treat and/or ameliorate a disease or disorder.
  • therapeutic compounds, agents, and moieties, as disclosed herein, may be biologics or small molecule therapeutics, or combinations thereof.
  • sustained release refers to a continuous or continual release of a therapeutic agent introduced into the body of a subject over a period of time and at a therapeutic level sufficient to achieve a desired therapeutic effect throughout the period of time.
  • the rate at which the therapeutic agent is released is slower than the rate of release of the therapeutic agent when it is administered alone or in a non- #13447332v2 sustained release formulation.
  • Sustained release formulations may, by way of example, be created as films, slabs, pellets, microparticles, microspheres, microcapsules, spheroids, shaped derivatives and paste.
  • the formulations may be in a form that is suitable for suspension in isotonic saline, physiological buffer or other solution acceptable for injection into a patient. Further, the formulations may be used in conjunction with any implantable, insertable or injectable system that a person of ordinary skill would appreciate as useful in connection with embodiments herein including but not limited to parenteral formulations, microspheres, microcapsules, gels, pastes, implantable rods, pellets, plates or fibers, etc.
  • 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.
  • 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. In certain embodiments, the particle is substantially solid throughout. In certain embodiments, the particle is a nanoparticle. In certain embodiments, 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).
  • FIG.1 shows a synthetic scheme of BNP@BSA-X, a glutaraldehyde-crosslinked bovine serum albumin hydrogel (BSA-X) containing bupivacaine free base nanoparticles (BNPs).
  • FIGs.2A-2E show characterization of BNP@BSA-X.
  • FIG.2A shows size distribution by volume percent of BSA and BNP@BSA measured by dynamic light scattering (DLS).
  • FIG. 2B shows Transmission electron micrograph (TEM) of BNP@BSA. Scale bar: 100nm.
  • FIGs.5A-5D show tissue retention of fluorescently labeled formulations injected at the sciatic nerve, detected by IVIS.
  • FIG.5A shows representative time courses of retention of ICG dye containing formulations
  • FIG.5B shows quantification of the fluorescence intensity over #13447332v2 time, derived from data such as those in FIG.5A.
  • FIG.5C shows representative time courses of retention of Cy5.5 containing formulations.
  • FIG.5D shows quantification of the fluorescence intensity over time, derived from data such as those in FIG.5C.
  • Bupivacaine concentration was 20 mg/ml.
  • p-values compare groups at 7 days.
  • FIGs.6A-6B show comparison of tissue retention and the duration of sensory nerve block for different formulations.
  • FIG.5A shows duration of block as a function of the half-life of tissue retention of ICG, calculated from data in FIG.5B.
  • FIG.6B shows Tissue retention of Cy5.5 on day 2, calculated from the data in FIG.5D.
  • FIGs.7A-7E show tissue reaction.
  • FIG.7A-7B Representative hematoxylin-eosin stained sections of muscles and nerve 4 days after injection of (FIGs.7A-7B) BNP@BSA-X with 20 mg/ml bupivacaine or (FIGs.7C-7D) EXPAREL ® .
  • Scale bar 200 ⁇ m (magnification 100x, FIGs.7A and 7C), 40 ⁇ m (magnification 400x, FIGs.7B and 7D).
  • M muscle
  • Mtox myotoxicity
  • N nerve
  • Infl inflammation
  • BSA-X crosslinked BSA.
  • FIG.8 shows a photograph of BNP, BNP@BSA and BNP@BSA-X. Ratios represent concentrations of BSA (mg/ml): GA (mg/ml). Bupivacaine concentration was set at 20 mg/ml. The cross-linked samples (BSA-X) formed hydrogels, as evidence by the fact that they did not flow down inverted tubes.
  • FIGs.13A-13C show DNP@BSA-X: A hydrogel derived from glutaraldehyde- crosslinked bovine serum albumin (BSA-X) encapsulating drug nanoparticles (DNPs).
  • FIG.13A #13447332v2 shows synthetic scheme for DNP@BSA-X incorporating either amphiphilic (diphenhydramine) or hydrophobic (paclitaxel) drugs.
  • FIG.13B shows transmission electron microscopy (TEM) images of DHNP@BSA (left panel; Scale bar: 100nm) and PNP@BSA (right panel; Scale bar: 500nm).
  • FIG.13C shows mechanical properties of DHNP@BSA-X and PNP@BSA-X determined by rheometry.
  • FIGS.18A-18B show representative hematoxylin-eosin stained sections of muscles 28 days after injection of BNP@BSA-X with 20 mg/ml bupivacaine.
  • FIG.18A Scale bar: 200 ⁇ m (magnification 100X)
  • FIG.18B Scale bar: 40 ⁇ m (magnification 400X).
  • FIG.19 shows thermal latency in uninjected animals (“uninjected”) and in the uninjected (contralateral) hind paw when each formulation was injected into the other leg. The graph shows the thermal latency at the representative time point of 1 hr post-injection. Thermal latencies of uninjected legs were measured throughout the study, and none of the formulations caused deficits in the uninjected hind paw during the entire study period.
  • bupivacaine was self-assembled in the hydrophobic free base form into #13447332v2 nanoparticles to maximize loading and slow release of the drug.
  • the bupivacaine nanoparticles were formed in the presence of bovine serum albumin (BSA) to enhance nanoparticle stability and prevent aggregation. Since BSA also binds hydrophobic drugs such as bupivacaine, it may contribute to slow release of the drug.
  • BSA bovine serum albumin
  • the present disclosure also provides a composition comprising a hydrogel and a protein- stabilized nanoparticle, wherein: the hydrogel comprises a crosslinked protein matrix; the protein-stabilized nanoparticle comprises a therapeutic agent or a diagnostic agent; and the protein-stabilized nanoparticle is embedded in the crosslinked protein matrix.
  • the present disclosure also provides a composition comprising a hydrogel and a nanoparticle, wherein: the hydrogel comprises a crosslinked protein matrix; the nanoparticle comprises an anesthetic; and the nanoparticle is embedded in the crosslinked protein matrix. #13447332v2 As described herein, the composition comprises a hydrogel and a protein-stabilized nanoparticle, or the composition comprises a hydrogel and a nanoparticle. In some embodiments, the composition comprises additional components.
  • the hydrogel comprises a crosslinked protein matrix.
  • the hydrogel comprises saline.
  • the hydrogel comprises a buffer.
  • the hydrogel comprises pH 7.4 phosphate-buffered saline (PBS).
  • PBS pH 7.4 phosphate-buffered saline
  • the hydrogel comprises a buffer with a pH of about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13, or about 14.
  • the hydrogel comprises a buffer with a pH of about 4, about 4.5, about 5, about 5.5, about 6, about 6.5, about 7, about 7.5, about 8, about 8.5, about 9, about 9.5, or about 10.
  • the hydrogel comprises a buffer with a pH of between about 5 and about 9, between about 5 and about 8.5, between about 5 and about 8, between about 5 and about 7.5, between about 5 and about 7, between about 5 and about 6.5, between about 5 and about 6, between about 5.5 and about 9, between about 5.5 and about 8.5, between about 5.5 and about 8, between about 5.5 and about 7.5, between about 5.5 and about 7, between about 5.5 and about 6.5, between about 5.5 and about 6, between about 6 and about 9, between about 6 and about 8.5, between about 6 and about 8, between about 6 and about 7.5, between about 6 and about 7, between about 6 and about 6.5, between about 6.5 and about 9, between about 6.5 and about 8.5, between about 6.5 and about 8, between about 6.5 and about 7.5, between about 6.5 and about 7, between about 7 and about 9, between about 7 and about 8.5, between about 7 and about 8, between about 7 and about 7.5, between about 7.5 and about 9, between about 7.5 and about 8.5, between about 7.5 and about 8, between about 8 between about 7 and
  • the hydrogel comprises a buffer with a pH of between about 4 and about 11, between about 5 and about 10, between about 6 and about 9, or between about 7 and about 8.
  • Crosslinked Protein Matrix In some embodiments, the crosslinked protein matrix comprises a crosslinked protein. In some embodiments, the crosslinked protein is crosslinked bovine serum albumin or crosslinked alpha-1-acid glycoprotein. In some embodiments, the crosslinked protein is crosslinked bovine #13447332v2 serum albumin. In some embodiments, the crosslinked protein is crosslinked alpha-1-acid glycoprotein. In some embodiments, the crosslinked protein matrix is non-covalently crosslinked. In some embodiments, the crosslinked protein matrix is covalently crosslinked.
  • the crosslinked protein is covalently crosslinked bovine serum albumin or covalently crosslinked alpha-1-acid glycoprotein. In some embodiments, the crosslinked protein is covalently crosslinked bovine serum albumin. In some embodiments, the crosslinked protein is covalently crosslinked alpha-1-acid glycoprotein.
  • the therapeutic agent e.g., bupivacaine
  • the therapeutic agent is capable of binding to the crosslinked protein. In some embodiments, the therapeutic agent (e.g., bupivacaine) is capable of binding to crosslinked bovine serum albumin or crosslinked alpha-1-acid glycoprotein. In some embodiments, the therapeutic agent (e.g., bupivacaine) is capable of binding to crosslinked bovine serum albumin.
  • the therapeutic agent e.g., bupivacaine
  • the crosslinked protein matrix comprises a moiety of Formula (I): wherein: each instance of Z is independently a protein in the crosslinked protein matrix; and L is optionally substituted alkylene or optionally substituted heteroalkylene.
  • at least one instance of Z is bovine serum albumin or alpha-1-acid glycoprotein.
  • at least one instance of Z is bovine serum albumin.
  • at least one instance of Z is alpha-1-acid glycoprotein.
  • each instance of Z is bovine serum albumin or alpha-1-acid glycoprotein.
  • each instance of Z is bovine serum albumin. In some embodiments, each instance of Z is alpha- 1-acid glycoprotein. In some embodiments, each instance of Z is a different instance of the protein in the crosslinked protein matrix.
  • L is optionally substituted alkylene. In some embodiments, L is optionally substituted C 1-12 alkylene. In some embodiments, L is optionally substituted C 1-6 alkylene. In some embodiments, L is optionally substituted C1-3 alkylene.
  • L is optionally substituted C12 alkylene, optionally substituted C11 alkylene, optionally substituted C 10 alkylene, optionally substituted C 9 alkylene, optionally #13447332v2 substituted C8 alkylene, optionally substituted C7 alkylene, optionally substituted C6 alkylene, optionally substituted C 5 alkylene, optionally substituted C 4 alkylene, optionally substituted C 3 alkylene, optionally substituted C 2 alkylene, or optionally substituted C 1 alkylene.
  • L is optionally substituted C12 alkylene.
  • L is optionally substituted C11 alkylene.
  • L is optionally substituted C10 alkylene.
  • L is optionally substituted C 9 alkylene. In some embodiments, L is optionally substituted C8 alkylene. In some embodiments, L is optionally substituted C7 alkylene. In some embodiments, L is optionally substituted C6 alkylene. In some embodiments, L is optionally substituted C 5 alkylene. In some embodiments, L is optionally substituted C 4 alkylene. In some embodiments, L is optionally substituted C3 alkylene. In some embodiments, L is optionally substituted C2 alkylene. In some embodiments, L is optionally substituted C1 alkylene. In some embodiments, L is substituted alkylene. In some embodiments, L is substituted C 1-12 alkylene.
  • L is substituted C12 alkylene, substituted C11 alkylene, substituted C 10 alkylene, substituted C 9 alkylene, substituted C 8 alkylene, substituted C 7 alkylene, substituted C6 alkylene, substituted C5 alkylene, substituted C4 alkylene, substituted C3 alkylene, substituted C2 alkylene, or substituted C1 alkylene.
  • L is substituted C12 alkylene.
  • L is substituted C 11 alkylene.
  • L is substituted C10 alkylene.
  • L is substituted C9 alkylene.
  • L is substituted C8 alkylene.
  • L is substituted C7 alkylene. In some embodiments, L is substituted C 6 alkylene. In some embodiments, L is substituted C 5 alkylene. In some embodiments, L is substituted C 4 alkylene. In some embodiments, L is substituted C3 alkylene. In some embodiments, L is substituted C2 alkylene. In some embodiments, L is substituted C1 alkylene. In some embodiments, L is unsubstituted alkylene. In some embodiments, L is unsubstituted C1-12 alkylene. In some embodiments, L is unsubstituted C1-6 alkylene. In some embodiments, L is unsubstituted C1-3 alkylene.
  • L is unsubstituted C12 alkylene, unsubstituted C 11 alkylene, unsubstituted C 10 alkylene, unsubstituted C 9 alkylene, unsubstituted C8 alkylene, unsubstituted C7 alkylene, unsubstituted C6 alkylene, unsubstituted C5 alkylene, unsubstituted C4 alkylene, unsubstituted C3 alkylene, unsubstituted C2 alkylene, or unsubstituted C 1 alkylene.
  • L is unsubstituted C 12 alkylene.
  • L is unsubstituted C 11 alkylene.
  • L is unsubstituted C 10 alkylene. In some embodiments, L is unsubstituted C9 alkylene. In some embodiments, L is unsubstituted C8 alkylene. In some embodiments, L is unsubstituted C7 alkylene. In some embodiments, L is unsubstituted C 6 alkylene. In some embodiments, L is unsubstituted C 5 alkylene. In some embodiments, L is unsubstituted C4 alkylene. In some embodiments, L is unsubstituted C3 alkylene. In some embodiments, L is unsubstituted C2 alkylene. In some embodiments, L is unsubstituted C 1 alkylene.
  • L is methylene, ethylene, n-propylene, isopropylene, n-butylene, tert-butylene, sec-butylene, isobutylene, n-pentylene, 3- pentanylene, amylene, neopentylene, 3-methylene-2-butanylene, tert-amylene, or n-hexylene.
  • L is methylene, ethylene, n-propylene, n-butylene, n-pentylene, or n- hexylene.
  • L is methylene.
  • L is ethylene.
  • L is n-propylene.
  • L is optionally substituted heteroalkylene containing at least one instance of .
  • L is optionally containing at least one instance of –NR N –, wherein each instance of R N 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, optionally substituted heteroaryl, or a nitrogen protecting group, or two occurrences of R N are joined together with their intervening atoms to form an optionally substituted heterocyclic ring or optionally substituted heteroaryl ring.
  • L is optionally substituted heteroalkylene containing at least 1 instance of –NH–. In some embodiments, L is optionally substituted heteroalkylene containing at least 1 instance of – N(optionally substituted alkyl)–. In some embodiments, L is optionally substituted heteroalkylene containing at least 1 instance of –N(nitrogen protecting group)–.
  • the crosslinked protein matrix comprises a moiety of Formula (I- a): wherein each instance of Z is protein matrix. In some embodiments, the crosslinked protein matrix comprises a moiety of Formula (I-a), wherein at least one instance of Z is bovine serum albumin or alpha-1-acid glycoprotein.
  • the crosslinked protein matrix comprises a moiety of Formula (I-a), wherein at least one instance of Z is bovine serum albumin. In some embodiments, the crosslinked protein matrix comprises a moiety of Formula (I-a), wherein at least one instance of Z is alpha-1-acid glycoprotein. In some embodiments, the crosslinked protein matrix comprises a moiety of Formula (I-a), wherein each instance of Z is bovine serum albumin or alpha-1-acid glycoprotein. In some embodiments, the crosslinked protein matrix comprises a moiety of Formula (I-a), #13447332v2 wherein each instance of Z is bovine serum albumin.
  • the crosslinked protein matrix comprises a moiety of Formula (I-a), wherein each instance of Z is alpha-1-acid glycoprotein.
  • the crosslinked protein matrix is prepared by reacting a protein with a crosslinking agent.
  • the protein is not treated with ethanol prior to reacting with the crosslinking agent.
  • the protein is not denatured prior to reacting with the crosslinking agent.
  • the protein is bovine serum albumin or alpha-1-acid glycoprotein.
  • the protein is bovine serum albumin.
  • the protein is alpha-1-acid glycoprotein.
  • the therapeutic agent e.g., bupivacaine
  • the therapeutic agent is capable of binding to the protein.
  • the therapeutic agent e.g., bupivacaine
  • the therapeutic agent is capable of binding to bovine serum albumin or alpha-1-acid glycoprotein.
  • the therapeutic agent e.g., bupivacaine
  • the therapeutic agent is capable of binding to bovine serum albumin.
  • the therapeutic agent is capable of binding to alpha-1-acid glycoprotein.
  • Crosslinking Agent In some embodiments, the crosslinking agent comprises at least two acyl groups. In some embodiments, the crosslinking agent comprises two acyl groups.
  • the crosslinking agent comprises at least two aldehyde groups. In some embodiments, the crosslinking agent comprises two aldehyde groups. In some embodiments, the crosslinking agent is glutaraldehyde. In some embodiments, the crosslinking agent comprises at least two ketone groups. In some embodiments, the crosslinking agent comprises two ketone groups. In some embodiments, the crosslinking agent is of Formula (II): (II), wherein L is optionally substituted alkylene or optionally substituted heteroalkylene. In some embodiments, L is optionally substituted alkylene. In some embodiments, L is optionally substituted C1-12 alkylene. In some embodiments, L is optionally substituted C1-6 alkylene.
  • L is optionally substituted C 1-3 alkylene. In some embodiments, L is optionally substituted C 12 alkylene, optionally substituted C 11 alkylene, optionally substituted C10 alkylene, optionally substituted C9 alkylene, optionally substituted C 8 alkylene, optionally substituted C 7 alkylene, optionally substituted C 6 alkylene, optionally substituted C 5 alkylene, optionally substituted C 4 alkylene, optionally substituted C 3 alkylene, optionally substituted C2 alkylene, or optionally substituted C1 alkylene. In some embodiments, L is optionally substituted C 12 alkylene. In some embodiments, L is optionally #13447332v2 substituted C11 alkylene.
  • L is optionally substituted C10 alkylene. In some embodiments, L is optionally substituted C 9 alkylene. In some embodiments, L is optionally substituted C 8 alkylene. In some embodiments, L is optionally substituted C 7 alkylene. In some embodiments, L is optionally substituted C6 alkylene. In some embodiments, L is optionally substituted C5 alkylene. In some embodiments, L is optionally substituted C4 alkylene. In some embodiments, L is optionally substituted C 3 alkylene. In some embodiments, L is optionally substituted C2 alkylene. In some embodiments, L is optionally substituted C1 alkylene. In some embodiments, L is substituted alkylene.
  • L is substituted C12 alkylene, substituted C11 alkylene, substituted C 10 alkylene, substituted C 9 alkylene, substituted C 8 alkylene, substituted C 7 alkylene, substituted C 6 alkylene, substituted C 5 alkylene, substituted C 4 alkylene, substituted C 3 alkylene, substituted C2 alkylene, or substituted C1 alkylene.
  • L is substituted C12 alkylene.
  • L is substituted C11 alkylene.
  • L is substituted C 10 alkylene.
  • L is substituted C 9 alkylene.
  • L is substituted C8 alkylene.
  • L is substituted C7 alkylene. In some embodiments, L is substituted C6 alkylene. In some embodiments, L is substituted C5 alkylene. In some embodiments, L is substituted C 4 alkylene. In some embodiments, L is substituted C3 alkylene. In some embodiments, L is substituted C2 alkylene. In some embodiments, L is substituted C1 alkylene. In some embodiments, L is unsubstituted alkylene. In some embodiments, L is unsubstituted C 1-12 alkylene. In some embodiments, L is unsubstituted C 1-6 alkylene. In some embodiments, L is unsubstituted C1-3 alkylene.
  • L is unsubstituted C12 alkylene, unsubstituted C11 alkylene, unsubstituted C10 alkylene, unsubstituted C9 alkylene, unsubstituted C 8 alkylene, unsubstituted C 7 alkylene, unsubstituted C 6 alkylene, unsubstituted C 5 alkylene, unsubstituted C4 alkylene, unsubstituted C3 alkylene, unsubstituted C2 alkylene, or unsubstituted C1 alkylene.
  • L is unsubstituted C12 alkylene.
  • L is unsubstituted C 11 alkylene.
  • L is unsubstituted C 10 alkylene. In some embodiments, L is unsubstituted C9 alkylene. In some embodiments, L is unsubstituted C8 alkylene. In some embodiments, L is unsubstituted C7 alkylene. In some embodiments, L is unsubstituted C 6 alkylene. In some embodiments, L is unsubstituted C 5 alkylene. In some embodiments, L is unsubstituted C 4 alkylene. In some embodiments, L is unsubstituted C3 alkylene. In some embodiments, L is unsubstituted C2 alkylene. In some embodiments, L is unsubstituted C1 alkylene.
  • L is methylene, ethylene, n-propylene, isopropylene, n-butylene, tert-butylene, sec-butylene, isobutylene, n-pentylene, 3- pentanylene, amylene, neopentylene, 3-methylene-2-butanylene, tert-amylene, or n-hexylene.
  • L is methylene, ethylene, n-propylene, n-butylene, n-pentylene, or n- hexylene.
  • L is methylene.
  • L is ethylene.
  • L is n-propylene.
  • L is n-butylene. In some embodiments, L is n-pentylene. In some embodiments, L is n-hexylene. In some embodiments, L is optionally substituted heteroalkylene. In some embodiments, L is optionally substituted C 1-12 heteroalkylene. In some embodiments, L is optionally substituted C1-6 heteroalkylene. In some embodiments, L is optionally substituted C1-3 heteroalkylene. #13447332v2 In some embodiments, L is optionally substituted heteroalkylene containing at least one instance of –O–. In some embodiments, L is optionally substituted heteroalkylene containing at least 1, 2, 3, 4, or 5 instances of .
  • L is optionally substituted heteroalkylene containing at least 1 instance of –NH–. In some embodiments, L is optionally substituted heteroalkylene containing at least 1 instance of – N(optionally substituted alkyl)–. In some embodiments, L is optionally substituted heteroalkylene containing at least 1 instance of –N(nitrogen protecting group)–. Additional Embodiments, bovine serum albumin (BSA) refers to a compound having CAS Registry Number 9048-46-8.
  • BSA is encoded by the bovine ALB gene, located on chromosome 6 (e.g., encoded by Ensembl ID NO: ENSBTAG00000017121, Chromosome 6: 88,484,961-88,503,334 forward strand).
  • ALB encodes a peptide that is represented by NCBI Reference Sequence NP_851335.1.
  • ALB encodes an mRNA comprising the sequence set forth in NCBI Reference Sequence NM_180992.2.
  • alpha-1-acid glycoprotein refers to a compound having CAS Registry Number 66455-27-4.
  • the bovine serum albumin is present in a concentration of between about 25 mg/mL and about 500 mg/mL, between about 25 mg/mL and about 250 #13447332v2 mg/mL, between about 25 mg/mL and about 200 mg/mL, between about 25 mg/mL and about 175 mg/mL, between about 25 mg/mL and about 150 mg/mL, between about 25 mg/mL and about 125 mg/mL, between about 25 mg/mL and about 112.5 mg/mL, between about 25 mg/mL and about 105 mg/mL, between about 25 mg/mL and about 100 mg/mL, between about 37.5 mg/mL and about 500 mg/mL, between about 37.5 mg/mL and about 250 mg/mL, between about 37.5 mg/mL and about 200 mg/mL, between about 37.5 mg/mL and about 175 mg/mL, between about 37.5 mg/mL and about 150 mg/mL, between about 37.5 mg/mL and about
  • the bovine serum albumin is present in a concentration of between about 25 mg/mL and about 500 mg/mL, between about 25 mg/mL and about 250 mg/mL, between about 25 mg/mL and about 200 mg/mL, between about 25 mg/mL and about 175 mg/mL, between about 25 mg/mL and about 150 mg/mL, between about 25 mg/mL and about 125 mg/mL, between about 25 mg/mL and about 112.5 mg/mL, between about 25 mg/mL and about 105 mg/mL, between about 25 mg/mL and about 100 mg/mL, between about 37.5 mg/mL and about 125 mg/mL, between about 37.5 mg/mL and about 112.5 mg/mL, between about 37.5 mg/mL and about 105 mg/mL, between about 37.5 mg/mL and about 100 mg/mL, between about 45 mg/mL and about 125 mg/mL, between about 45 mg/mL and about 112.5 mg/mL
  • the bovine serum albumin is present in a concentration of between about 25 mg/mL and about 125 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of between about 37.5 mg/mL and about 112.5 mg/mL. In #13447332v2 some embodiments, the bovine serum albumin is present in a concentration of between about 45 mg/mL and about 105 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of between about 50 mg/mL and about 100 mg/mL.
  • the bovine serum albumin is present in a concentration of about 25 mg/mL, about 30 mg/mL, about 35 mg/mL, about 37.5 mg/mL, about 40 mg/mL, about 42.5 mg/mL, about 45 mg/mL, about 47.5 mg/mL, about 48 mg/mL, about 49 mg/mL, about 50 mg/mL, about 51 mg/mL, about 52 mg/mL, about 52.5 mg/mL, about 55 mg/mL, about 57.5 mg/mL, about 60 mg/mL, about 62.5 mg/mL, about 65 mg/mL, about 70 mg/mL, about 75 mg/mL, about 80 mg/mL about 85 mg/mL, about 87.5 mg/mL, about 90 mg/mL, about 92.5 mg/mL, about 95 mg/mL, about 97.5 mg/mL, about 98 mg/mL, about 99 mg/mL, about 100 mg/mL, about 101 mg/mL, about 102 mg/mL,
  • the bovine serum albumin is present in a concentration of about 35 mg/mL, about 37.5 mg/mL, about 40 mg/mL, about 42.5 mg/mL, about 45 mg/mL, about 47.5 mg/mL, about 48 mg/mL, about 49 mg/mL, about 50 mg/mL, about 51 mg/mL, about 52 mg/mL, about 52.5 mg/mL, about 55 mg/mL, about 57.5 mg/mL, about 60 mg/mL, about 62.5 mg/mL, or about 65 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 35 mg/mL.
  • the bovine serum albumin is present in a concentration of about 50 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 51 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 52 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 52.5 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about #13447332v2 55 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 57.5 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 60 mg/mL.
  • the glutaraldehyde is present in a concentration of between about 1 mg/mL and about 25 mg/mL, between about 1 mg/mL and about 20 mg/mL, between about 1 mg/mL and about 15 mg/mL, between about 1 mg/mL and about 14 mg/mL, between about 1 mg/mL and about 13 mg/mL, between about 1 mg/mL and about 12.5 mg/mL, between about 1 mg/mL and about 12 mg/mL, between about 1 mg/mL and about 11.5 mg/mL, between about 1 mg/mL and about 11 mg/mL, between about 1 mg/mL and about 10.5 mg/mL, between about 1 mg/mL and about 10 mg/mL, between about 2.5 mg/mL and about 25 mg/mL, between about #13447332v2 2.5 mg/mL and about 20 mg/mL, between about 2.5 mg/mL and about 15 mg/mL, between about 2.5 mg/mL and about 14 mg/mL,
  • the glutaraldehyde is present in a concentration of between about 1 mg/mL and about 15 mg/mL. In some embodiments, the glutaraldehyde is present in a concentration of between about 2.5 mg/mL and about 12.5 mg/mL. In some embodiments, the glutaraldehyde is present in a concentration of between about 4 mg/mL and about 11 mg/mL. In some embodiments, the glutaraldehyde is present in a concentration of between about 4.5 mg/mL and about 10.5 mg/mL. In some embodiments, the glutaraldehyde is present in a concentration of between about 5 mg/mL and about 10 mg/mL.
  • the glutaraldehyde is present in a concentration of about 1 mg/mL, about 1.5 mg/mL, about 2 mg/mL, about 2.5 mg/mL, about 3 mg/mL, about 3.5 mg/mL, about 4 mg/mL, about 4.5 mg/mL, about 5 mg/mL, about 5.5 mg/mL, about 6 mg/mL, about 6.5 mg/mL, about 7 mg/mL, about 7.5 mg/mL, about 8 mg/mL, about 8.5 mg/mL, about 9 mg/mL, about 9.5 mg/mL, about 10 mg/mL, about 10.5 mg/mL, about 11 mg/mL, about 11.5 mg/mL, about 12 mg/mL, about 12.5 mg/mL, about 13 mg/mL, about 13.5 mg/mL, about 14 mg/mL, about 14.5 mg/mL, about 15 mg/mL, about 16 mg/mL, about 17 mg/mL, about 18 mg/mL
  • the bovine serum albumin is present in a concentration of between about 25 mg/mL and about 125 mg/mL, and the glutaraldehyde is present in a concentration of between about 4 mg/mL and about 11 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of between about 25 mg/mL and about 125 mg/mL, and the glutaraldehyde is present in a concentration of between about 4.5 mg/mL and about 10.5 mg/mL.
  • the bovine serum albumin is present in a concentration of between about 37.5 mg/mL and about 112.5 mg/mL, and the glutaraldehyde is present in a concentration of between about 4.5 mg/mL and about 10.5 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of between about 37.5 mg/mL and about 112.5 mg/mL, and the glutaraldehyde is present in a concentration of between about 5 mg/mL and about 10 mg/mL.
  • the bovine serum albumin is present in a concentration of between about 45 mg/mL and about 105 mg/mL, and the glutaraldehyde is present in a concentration of between about 5 mg/mL and about 10 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of between about 50 mg/mL and about 100 mg/mL, and the glutaraldehyde is present in a concentration of between about 1 mg/mL and about 15 mg/mL.
  • the bovine serum albumin is present in a concentration of between about 50 mg/mL and about 100 mg/mL, and the glutaraldehyde is present in a concentration of between about 2.5 mg/mL and about 12.5 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of between about 50 mg/mL and about 100 mg/mL, and the glutaraldehyde is present in a concentration of between about 4 mg/mL and about 11 mg/mL.
  • the bovine serum albumin is present in a concentration of between about 50 mg/mL and about 100 mg/mL, and the glutaraldehyde is present in a concentration of between about 4.5 mg/mL and about 10.5 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of between about 50 mg/mL and about 100 mg/mL, and the glutaraldehyde is present in a concentration of between about 5 mg/mL and about 10 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 40 mg/mL, and the glutaraldehyde is present in a concentration of about 4 mg/mL.
  • the bovine serum albumin is present in a concentration of about 45 mg/mL, and the glutaraldehyde is present in a concentration of about 5.5 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 45 mg/mL, and the glutaraldehyde is present in a concentration of about 6 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 45 mg/mL, and the glutaraldehyde is present in a concentration of about 9 mg/mL.
  • the bovine serum albumin is present in a concentration of about 55 mg/mL, and the glutaraldehyde is present in a concentration of about 5.5 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 55 mg/mL, and the glutaraldehyde is present in a concentration of about 6 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 55 mg/mL, and the glutaraldehyde is present in a concentration of about 9 mg/mL.
  • the bovine serum albumin is present in a concentration of about 55 mg/mL, and the glutaraldehyde is present in a concentration of about 9.5 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 55 mg/mL, and the glutaraldehyde is present in a concentration of about 10 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 55 mg/mL, and the glutaraldehyde is present in a concentration of about 10.5 mg/mL.
  • the bovine serum albumin is present in a concentration of about 60 mg/mL, and the glutaraldehyde #13447332v2 is present in a concentration of about 5 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 60 mg/mL, and the glutaraldehyde is present in a concentration of about 5.5 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 60 mg/mL, and the glutaraldehyde is present in a concentration of about 6 mg/mL.
  • the bovine serum albumin is present in a concentration of about 60 mg/mL, and the glutaraldehyde is present in a concentration of about 9 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 60 mg/mL, and the glutaraldehyde is present in a concentration of about 9.5 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 60 mg/mL, and the glutaraldehyde is present in a concentration of about 10 mg/mL.
  • the bovine serum albumin is present in a concentration of about 60 mg/mL, and the glutaraldehyde is present in a concentration of about 10.5 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 60 mg/mL, and the glutaraldehyde is present in a concentration of about 11 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 90 mg/mL, and the glutaraldehyde is present in a concentration of about 4 mg/mL.
  • the bovine serum albumin is present in a concentration of about 90 mg/mL, and the glutaraldehyde is present in a concentration of about 4.5 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 90 mg/mL, and the glutaraldehyde is present in a concentration of about 5 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 90 mg/mL, and the glutaraldehyde is present in a concentration of about 5.5 mg/mL.
  • the bovine serum albumin is present in a concentration of about 90 mg/mL, and the glutaraldehyde is present in a concentration of about 6 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 90 mg/mL, and the glutaraldehyde is present in a concentration of about 9 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 90 mg/mL, and the glutaraldehyde is present in a concentration of about 9.5 mg/mL.
  • the bovine serum albumin is present in a concentration of about 90 mg/mL, and the glutaraldehyde is present in a concentration of about 10 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 90 mg/mL, and the glutaraldehyde is present in a concentration of about 10.5 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 90 mg/mL, and the glutaraldehyde is present in a concentration of about 11 mg/mL.
  • the bovine serum albumin is present in a concentration of about 95 mg/mL, and the glutaraldehyde is present in a concentration of about 5.5 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 95 mg/mL, and the glutaraldehyde is present in a concentration of about 6 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 95 mg/mL, and the glutaraldehyde is present in a concentration of about 9 mg/mL.
  • the bovine serum albumin is present in a concentration of about 95 mg/mL, and the glutaraldehyde is present in a concentration of about 9.5 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 95 mg/mL, and the glutaraldehyde is present in a concentration of about 10 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 95 mg/mL, and the glutaraldehyde is present in a concentration of about 10.5 mg/mL.
  • the bovine serum albumin is present in a concentration of about 95 mg/mL, and the glutaraldehyde is present in a concentration of about 11 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 97.5 mg/mL, and the glutaraldehyde is present in a concentration of about 4 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 97.5 mg/mL, and the glutaraldehyde is present in a concentration of about 4.5 mg/mL.
  • the bovine serum albumin is present in a concentration of about 97.5 mg/mL, and the glutaraldehyde is present in a concentration of about 5 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 97.5 mg/mL, and the glutaraldehyde is present in a concentration of about 5.5 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 97.5 mg/mL, and the glutaraldehyde is present in a concentration of about 6 mg/mL.
  • the bovine serum albumin is present in a concentration of about 97.5 mg/mL, and the glutaraldehyde is present in a concentration of about 9 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 97.5 mg/mL, and the glutaraldehyde is present in a concentration of about 9.5 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 97.5 mg/mL, and the glutaraldehyde is present in a concentration of about 10 mg/mL.
  • the bovine serum albumin is present in a concentration of about 97.5 mg/mL, and the glutaraldehyde is present in a concentration of about 10.5 mg/mL. In some embodiments, the bovine serum #13447332v2 albumin is present in a concentration of about 97.5 mg/mL, and the glutaraldehyde is present in a concentration of about 11 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 100 mg/mL, and the glutaraldehyde is present in a concentration of about 4 mg/mL.
  • the bovine serum albumin is present in a concentration of about 100 mg/mL, and the glutaraldehyde is present in a concentration of about 4.5 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 100 mg/mL, and the glutaraldehyde is present in a concentration of about 5 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 100 mg/mL, and the glutaraldehyde is present in a concentration of about 5.5 mg/mL.
  • the bovine serum albumin is present in a concentration of about 100 mg/mL, and the glutaraldehyde is present in a concentration of about 6 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 100 mg/mL, and the glutaraldehyde is present in a concentration of about 9 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 100 mg/mL, and the glutaraldehyde is present in a concentration of about 9.5 mg/mL.
  • the bovine serum albumin is present in a concentration of about 100 mg/mL, and the glutaraldehyde is present in a concentration of about 10 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 100 mg/mL, and the glutaraldehyde is present in a concentration of about 10.5 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 100 mg/mL, and the glutaraldehyde is present in a concentration of about 11 mg/mL.
  • the bovine serum albumin is present in a concentration of about 102.5 mg/mL, and the glutaraldehyde is present in a concentration of about 4 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 102.5 mg/mL, and the glutaraldehyde is present in a concentration of about 4.5 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 102.5 mg/mL, and the glutaraldehyde is present in a concentration of about 5 mg/mL.
  • the bovine serum albumin is present in a concentration of about 102.5 mg/mL, and the glutaraldehyde is present in a concentration of about 5.5 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 102.5 mg/mL, and the glutaraldehyde is present in a concentration of about 6 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 102.5 mg/mL, and the glutaraldehyde is present in a concentration of about 9 mg/mL.
  • the bovine serum albumin is present in a concentration of about 102.5 mg/mL, and the glutaraldehyde is present in a concentration of about 9.5 mg/mL.
  • the #13447332v2 bovine serum albumin is present in a concentration of about 102.5 mg/mL, and the glutaraldehyde is present in a concentration of about 10 mg/mL.
  • the bovine serum albumin is present in a concentration of about 102.5 mg/mL, and the glutaraldehyde is present in a concentration of about 10.5 mg/mL.
  • the bovine serum albumin is present in a concentration of about 102.5 mg/mL, and the glutaraldehyde is present in a concentration of about 11 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 105 mg/mL, and the glutaraldehyde is present in a concentration of about 4 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 105 mg/mL, and the glutaraldehyde is present in a concentration of about 4.5 mg/mL.
  • the bovine serum albumin is present in a concentration of about 105 mg/mL, and the glutaraldehyde is present in a concentration of about 5 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 105 mg/mL, and the glutaraldehyde is present in a concentration of about 5.5 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 105 mg/mL, and the glutaraldehyde is present in a concentration of about 6 mg/mL.
  • the bovine serum albumin is present in a concentration of about 105 mg/mL, and the glutaraldehyde is present in a concentration of about 9 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 105 mg/mL, and the glutaraldehyde is present in a concentration of about 9.5 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 105 mg/mL, and the glutaraldehyde is present in a concentration of about 10 mg/mL.
  • the bovine serum albumin is present in a concentration of about 105 mg/mL, and the glutaraldehyde is present in a concentration of about 10.5 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 105 mg/mL, and the glutaraldehyde is present in a concentration of about 11 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 110 mg/mL. the glutaraldehyde is present in a concentration of about 4 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 110 mg/mL.
  • the glutaraldehyde is present in a concentration of about 4.5 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 110 mg/mL. the glutaraldehyde is present in a concentration of about 5 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 110 mg/mL. the glutaraldehyde is present in a concentration of about 5.5 mg/mL. In some embodiments, the bovine serum albumin is present in a concentration of about 110 mg/mL. the glutaraldehyde is present in a concentration of about 6 mg/mL.
  • the bovine serum albumin is present in a concentration of #13447332v2 about 110 mg/mL.
  • the glutaraldehyde is present in a concentration of about 9 mg/mL.
  • the bovine serum albumin is present in a concentration of about 110 mg/mL.
  • the glutaraldehyde is present in a concentration of about 9.5 mg/mL.
  • the bovine serum albumin is present in a concentration of about 110 mg/mL.
  • the glutaraldehyde is present in a concentration of about 10 mg/mL.
  • the bovine serum albumin is present in a concentration of about 110 mg/mL.
  • the glutaraldehyde is present in a concentration of about 10.5 mg/mL.
  • the bovine serum albumin is present in a concentration of about 110 mg/mL.
  • the glutaraldehyde is present in a concentration of about 11 mg/mL.
  • Nanoparticle As described herein, the nanoparticle comprises a therapeutic agent or a diagnostic agent; and the nanoparticle is embedded in the crosslinked protein matrix.
  • the nanoparticle consists of a therapeutic agent or a diagnostic agent, and the nanoparticle is embedded in the crosslinked protein matrix.
  • the nanoparticle consists essentially of a therapeutic agent or a diagnostic agent, and the nanoparticle is embedded in the crosslinked protein matrix.
  • the nanoparticle does not comprise a protein. In some embodiments, the nanoparticle does not comprise a crosslinked protein. In some embodiments, the nanoparticle does not comprise the crosslinked protein matrix. In some embodiments, the nanoparticle does not comprise the hydrogel. In some embodiments, the nanoparticle does not comprise water. In some embodiments, the nanoparticle does not comprise bovine serum albumin. In some embodiments, the nanoparticle does not comprise a crosslinking agent. In some embodiments, the nanoparticle does not comprise glutaraldehyde.
  • the nanoparticle comprises a hydrodynamic diameter of between about 100 nm and about 400 nm, between about 100 nm and about 300 nm, between about 100 nm and about 200 nm, between about 200 nm and about 300 nm, between about 125 nm and about 300 nm, between about 125 nm and about 275 nm, between about 125 nm and about 250 nm, between about 125 nm and about 225 nm, between about 150 nm and about 300 nm, between about 150 nm and about 275 nm, between about 150 nm and about 250 nm, between about 150 nm and about 225 nm, between about 175 nm and about 300 nm, between about 175 nm and about 275 nm, between about 175 nm and about 250 nm, or between about 175 nm and about 225 nm.
  • the nanoparticle comprises a hydrodynamic diameter of about 100 nm, about 110 nm, about 120 nm, about 130 nm, about 140 nm, about 150 nm, about 160 #13447332v2 nm, about 170 nm, about 175 nm, about 180 nm, about 185 nm, about 190 nm, about 195 nm, about 200 nm, about 205 nm, about 210 nm, about 215 nm, about 220 nm, about 225 nm, about 230 nm, about 240 nm, about 250 nm, about 260 nm, about 270 nm, about 280 nm, about 290 nm, about 300 nm, about 310 nm, about 320 nm, about 330 nm, about 340 nm, about 350 nm, about 360 nm, about 370 nm, about 380 nm,
  • the nanoparticle comprises a hydrodynamic diameter of about 175 nm. In some embodiments, the nanoparticle comprises a hydrodynamic diameter of about 180 nm. In some embodiments, the nanoparticle comprises a hydrodynamic diameter of about 185 nm. In some embodiments, the nanoparticle comprises a hydrodynamic diameter of about 190 nm. In some embodiments, the nanoparticle comprises a hydrodynamic diameter of about 195 nm. In some embodiments, the nanoparticle comprises a hydrodynamic diameter of about 200 nm. In some embodiments, the nanoparticle comprises a hydrodynamic diameter of about 205 nm. In some embodiments, the nanoparticle comprises a hydrodynamic diameter of about 210 nm.
  • the nanoparticle comprises a hydrodynamic diameter of about 215 nm. In some embodiments, the nanoparticle comprises a hydrodynamic diameter of about 220 nm. In some embodiments, the nanoparticle comprises a hydrodynamic diameter of about 225 nm. In some embodiments, the nanoparticle is covalently attached to the hydrogel. In some embodiments, the nanoparticle is not covalently attached to the hydrogel. In some embodiments, the nanoparticle is covalently attached to the crosslinked protein matrix. In some embodiments, the nanoparticle is not covalently attached to the crosslinked protein matrix. In some embodiments, the nanoparticle is not covalently attached to the hydrogel and/or the nanoparticle is not covalently attached to the crosslinked protein matrix.
  • the nanoparticle is not covalently attached to the hydrogel, and the nanoparticle is not covalently attached to the crosslinked protein matrix.
  • Protein-Stabilized Nanoparticle As described herein, the protein-stabilized nanoparticle comprises a therapeutic agent or a diagnostic agent; and the protein-stabilized nanoparticle is embedded in the crosslinked protein matrix. In some embodiments, the protein-stabilized nanoparticle consists of a therapeutic agent or a diagnostic agent, and the protein-stabilized nanoparticle is embedded in the crosslinked protein matrix. In some embodiments, the protein-stabilized nanoparticle consists essentially of a therapeutic agent or a diagnostic agent, and the protein-stabilized nanoparticle is embedded in the crosslinked protein matrix.
  • the protein-stabilized nanoparticle does not comprise a protein. In some embodiments, the protein-stabilized nanoparticle does not comprise a crosslinked protein. In some embodiments, the protein-stabilized nanoparticle does not comprise the crosslinked protein matrix. In some embodiments, the protein-stabilized nanoparticle does not comprise the hydrogel. In some embodiments, the protein-stabilized nanoparticle does not comprise water. In some embodiments, the protein-stabilized nanoparticle does not comprise bovine serum albumin. In some embodiments, the protein-stabilized nanoparticle does not comprise a crosslinking agent. In some embodiments, the protein-stabilized nanoparticle does not comprise glutaraldehyde.
  • the crosslinked protein matrix comprises a crosslinked protein
  • the protein-stabilized nanoparticle comprises a therapeutic agent (e.g., bupivacaine) capable of binding to the crosslinked protein.
  • the crosslinked protein matrix comprises crosslinked bovine serum albumin or crosslinked alpha-1-acid glycoprotein
  • the protein-stabilized nanoparticle comprises a therapeutic agent (e.g., bupivacaine) capable of binding to the crosslinked bovine serum albumin or crosslinked alpha-1-acid glycoprotein.
  • the crosslinked protein matrix is prepared by reacting bovine serum albumin or alpha-1-acid glycoprotein with a crosslinking agent, and the protein-stabilized nanoparticle comprises a therapeutic agent (e.g., bupivacaine) capable of binding to the bovine serum albumin or alpha-1-acid glycoprotein.
  • the crosslinked protein matrix is prepared by reacting bovine serum albumin with a crosslinking agent, and the protein-stabilized nanoparticle comprises a therapeutic agent (e.g., bupivacaine) capable of binding to the bovine serum albumin.
  • the crosslinked protein matrix is prepared by reacting alpha-1-acid glycoprotein with a crosslinking agent, and the protein-stabilized nanoparticle comprises a therapeutic agent (e.g., bupivacaine) capable of binding to the alpha-1-acid glycoprotein.
  • a therapeutic agent e.g., bupivacaine
  • the protein-stabilized nanoparticle comprises a hydrodynamic diameter of about 175 nm. In some embodiments, the protein-stabilized nanoparticle comprises a hydrodynamic diameter of about 180 nm. In some embodiments, the protein-stabilized nanoparticle comprises a hydrodynamic diameter of about 185 nm. In some embodiments, the protein-stabilized nanoparticle comprises a hydrodynamic diameter of about 190 nm. In some embodiments, the protein-stabilized nanoparticle comprises a hydrodynamic diameter of about 195 nm. In some embodiments, the protein-stabilized nanoparticle comprises a hydrodynamic diameter of about 200 nm. In some embodiments, the protein-stabilized nanoparticle comprises a hydrodynamic diameter of about 205 nm.
  • the protein-stabilized nanoparticle comprises a hydrodynamic diameter of about 210 nm. In some embodiments, the protein-stabilized nanoparticle comprises a hydrodynamic diameter of about 215 nm. In some embodiments, the protein-stabilized nanoparticle comprises a hydrodynamic diameter of about 220 nm. In some embodiments, the protein-stabilized nanoparticle comprises a hydrodynamic diameter of about 225 nm. In some embodiments, the protein-stabilized nanoparticle is covalently attached to the hydrogel. In some embodiments, the protein-stabilized nanoparticle is not covalently attached to the hydrogel. In some embodiments, the protein-stabilized nanoparticle is covalently attached to the crosslinked protein matrix.
  • the protein-stabilized nanoparticle is not covalently attached to the crosslinked protein matrix. In some embodiments, the protein- stabilized nanoparticle is not covalently attached to the hydrogel and/or the protein-stabilized nanoparticle is not covalently attached to the crosslinked protein matrix. In some embodiments, #13447332v2 the protein-stabilized nanoparticle is not covalently attached to the hydrogel, and the protein- stabilized nanoparticle is not covalently attached to the crosslinked protein matrix. Diagnostic Agent In some embodiments, the protein-stabilized nanoparticle comprises a diagnostic agent. In some embodiments, the diagnostic agent is a fluorophore. In some embodiments, the diagnostic agent is a small molecule or a biologic.
  • the diagnostic agent is a small molecule. In some embodiments, the diagnostic agent is conjugated to a protein, a polymer, or a small molecule. In some embodiments, the diagnostic agent is conjugated to the protein-stabilized nanoparticle. In some embodiments, the diagnostic agent is not conjugated to the protein-stabilized nanoparticle. In some embodiments, the diagnostic agent is Sulforhodamine B, Cy-5.5, indocyanine green, fluorescein isothiocyanate, methylene blue, or coumarin. In some embodiments, the diagnostic agent comprises, by weight, about 0.05% to about 10% of the total mass of the composition.
  • the diagnostic agent comprises, by weight, about 0.05% to about 0.2%, about 0.1% to about 0.2%, about 0.12% to about 0.2%, about 0.13% to about 0.2%, about 0.14% to about 0.2%, about 0.15% to about 0.2%, about 0.15%, or about 0.16% of the total mass of the composition.
  • the diagnostic agent comprises, by weight, about 2% to about 4%, about 2.5% to about 3.5%, about 2% to about 3%, about 2.8% to about 3.2%, about 2.8% to about 3.0%, about 2.8%, about 2.9%, about 3.0%, about 3.1%, or about 0.16% of the total mass of the composition.
  • the therapeutic agent does not comprise a primary or secondary amine. In some embodiments, the therapeutic agent is not doxorubicin. In some embodiments, the therapeutic agent is hydrophobic. In some embodiments, the therapeutic agent is hydrophilic. In some embodiments, the therapeutic agent is amphiphilic. In certain embodiments, the therapeutic agent has a LogD or LogP that is equal to or less than 0. In certain embodiments, the therapeutic agent has a LogD or LogP that is equal to or less than 0.5. In certain embodiments, the therapeutic agent has a LogD or LogP that is equal to or less than 1. In certain embodiments, the therapeutic agent has a LogD or LogP that is equal to or less than 1.5.
  • the therapeutic agent has a LogD or LogP that is equal to or less than 2. In certain embodiments, the therapeutic agent has a LogD or LogP that is equal to or less than 2.5. In certain embodiments, the therapeutic agent has a LogD or LogP that is equal to or less than 3. In certain embodiments, the therapeutic agent has a LogD or LogP that is equal to or less than 3.5. In certain embodiments, the therapeutic agent has a LogD or LogP that is equal to or less than 4. In certain embodiments, the therapeutic agent has a LogD or LogP that is equal to or less than 4.5. In certain embodiments, the therapeutic agent has a LogD or LogP that is equal to or less than 5.
  • the therapeutic agent has a LogD or LogP that is equal to or less than -0.5. In certain embodiments, the therapeutic agent has a LogD or LogP that is equal to or less than -1. In certain embodiments, the therapeutic agent has a LogD or LogP that is equal to or less than -1.5. In certain embodiments, the therapeutic agent has a LogD or LogP that is equal to or less than -2. In certain embodiments, the therapeutic agent has a LogD or LogP that is equal to or less than -2.5. In certain embodiments, the therapeutic agent has a LogD or LogP that is equal to or less than -3. In certain embodiments, the therapeutic agent has a LogD or LogP that is equal to or less than -3.5.
  • the therapeutic agent has a LogD or LogP that is equal to or less than -4. In certain embodiments, the therapeutic agent has a LogD or LogP that is equal to or less than -4.5. In certain embodiments, the therapeutic agent has a LogD or LogP that is equal to or less than -5. In certain embodiments, the therapeutic agent has a LogD or LogP that is -5 to 0. In certain embodiments, the therapeutic agent has a LogD or LogP that is 0 to 5.
  • the therapeutic agent is a local anesthetic (e.g., tetrodotoxin, saxitoxin, neosaxitoxin, bupivacaine, amylocaine, ambucaine, articaine, benzocaine, benzonatate, butacaine, butanilicaine, carbocaine, cepastat, chloraseptic, chloroprocaine, cinchocaine, citanest, cyclomethycaine, dibucaine, diperodon, dimethocaine, eucaine, #13447332v2 etidocaine, fomocaine, fotocaine, hydroxyprocaine, isobucaine, levobupivacaine, lidocaine, marcaine, mepivacaine, meprylcaine, metabutoxycaine, nitracaine, orthocaine, orabloc, oxetacaine, oxybuprocaine, paraethoxyca
  • the therapeutic agent is a free base form of a local anesthetic.
  • the therapeutic agent is a local anesthetic compound, for example, but not limited to, an amino ester compound (e.g., procaine, tetracaine, chloroprocaine, benzocaine, butacaine, dimethocaine) or an amino amide compound (e.g., procainamide, lidocaine).
  • the local anesthetic is a sodium channel blocker, for example, a site 1 sodium channel blocker (e.g., tetrodotoxin, saxitoxins (saxitoxin, neosaxitoxin), gonyautoxins (gonyautoxin V, gonyautoxin VI), ⁇ - conotoxins)) or an amino amide local anesthetic.
  • the therapeutic agent is a site 1 sodium channel blocker, amino ester anesthetic, or an amino amide anesthetic, or a derivative thereof.
  • the therapeutic agent is tetrodotoxin, saxitoxin, neosaxitoxin, bupivacaine, amylocaine, ambucaine, articaine, benzocaine, benzonatate, butacaine, butanilicaine, carbocaine, cepastat, chloraseptic, chloroprocaine, cinchocaine, citanest, cyclomethycaine, dibucaine, diperodon, dimethocaine, eucaine, etidocaine, fomocaine, fotocaine, hydroxyprocaine, isobucaine, levobupivacaine, lidocaine, marcaine, mepivacaine, meprylcaine, metabutoxycaine, nitracaine, orthocaine, orabloc, oxetacaine, oxybuprocaine, paraethoxycaine, phenacaine, piperocaine, piridocaine, poloc
  • the therapeutic agent is bupivacaine. In some embodiments, the therapeutic agent is a free base form of bupivacaine. In some embodiments, the therapeutic agent is an anti-cancer agent (e.g., anti-estrogens (e.g., tamoxifen, raloxifene, and megestrol), LHRH agonists (e.g., goscrclin and leuprolide), anti-androgens (e.g., flutamide and bicalutamide), photodynamic therapies (e.g., vertoporfin (BPD-MA), phthalocyanine, photosensitizer Pc4, and demethoxy-hypocrellin A (2BA-2- DMHA)), nitrogen mustards (e.g., cyclophosphamide, ifosfamide, trofosfamide, chlorambucil, estramustine, and melphalan), nitrosoureas (e.g., car
  • the therapeutic agent is a taxoid (e.g., paclitaxel or a paclitaxel equivalent such as nanoparticle albumin-bound paclitaxel (Abraxane), docosahexaenoic acid bound-paclitaxel (DHA-paclitaxel, Taxoprexin), polyglutamate bound-paclitaxel (PG-paclitaxel, paclitaxel poliglumex, CT-2103, XYOTAX), the tumor-activated prodrug (TAP) ANG1005 (Angiopep-2 bound to three molecules of paclitaxel), paclitaxel-EC-1 (paclitaxel bound to the erbB2- recognizing peptide EC-1), or glucose-conjugated paclitaxel, e.g., ’2’-paclitaxel methyl 2- glucopyranosyl succinate; docetaxel, taxol).
  • a taxoid e.g., paclitaxe
  • the therapeutic agent is a free base form of an anti-cancer agent.
  • the anti-cancer agent is not doxorubicin.
  • the therapeutic agent is paclitaxel. In some embodiments, the therapeutic agent is a free base form of paclitaxel.
  • the therapeutic agent is an antihistamine (e.g., diphenhydramine, chlorpheniramine, cetirizine, loratadine, fexofenadine, levocetirizine, desloratadine, Brompheniramine, Clemastine, Cyproheptadine, Dexchlorpheniramine Dimenhydrinate, Doxylamine Hydroxyzine, Phenindamine, Azelastine, Cimetidine, Famotidine, Nizatidine, or Ranitidine).
  • an antihistamine e.g., diphenhydramine, chlorpheniramine, cetirizine, loratadine, fexofenadine, levocetirizine, desloratadine, Brompheniramine, Clemastine, Cyproheptadine, Dexchlorpheniramine Dimenhydrinate, Doxylamine Hydroxyzine, Phenindamine, Azelastine, Cimetidine, Famotidine, Nizatidine
  • the therapeutic agent is diphenhydramine, chlorpheniramine, cetirizine, loratadine, fexofenadine, levocetirizine, desloratadine, Brompheniramine, Clemastine, Cyproheptadine, Dexchlorpheniramine Dimenhydrinate, Doxylamine Hydroxyzine, Phenindamine, Azelastine, Cimetidine, Famotidine, Nizatidine, or Ranitidine.
  • the therapeutic agent is a free base form of an antihistamine.
  • the therapeutic agent is diphenhydramine.
  • the therapeutic agent is a free base form of diphenhydramine.
  • the amount of the therapeutic agent in the composition is about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 21%, about 22%, about 23%, about 24%, about 25%, about 26%, about 27%, about 28%, about 29%, about 30%, about 40%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, or about 100% greater than the amount of the therapeutic agent in a composition that is not a composition disclosed herein.
  • the therapeutic agent is bupivacaine present in the composition in a concentration of about 15 mg/mL. In some embodiments, the therapeutic agent is bupivacaine present in the composition in a concentration of about 17.5 mg/mL. In some embodiments, the therapeutic agent is bupivacaine present in the composition in a concentration of about 20 mg/mL. In some embodiments, the therapeutic agent is bupivacaine present in the composition in a concentration of about 22.5 mg/mL. In some embodiments, the therapeutic agent is bupivacaine present in the composition in a concentration of about 25 mg/mL.
  • the therapeutic agent is bupivacaine, the crosslinking agent is glutaraldehyde, and the protein is bovine serum albumin. In some embodiments, the therapeutic agent is bupivacaine, the crosslinking agent is glutaraldehyde, and the protein is alpha-1-acid glycoprotein. In some embodiments, the therapeutic agent is a free base form of bupivacaine, the crosslinking agent is of Formula (II), and the protein is bovine serum albumin or alpha-1-acid glycoprotein. In some embodiments, the therapeutic agent is a free base form of bupivacaine, the crosslinking agent is of Formula (II), and the protein is bovine serum albumin.
  • the composition is characterized in that, when administered to a subject, less than 100%, less than 90%, less than 80%, less than 70%, less than 60%, less than 50%, less than 40%, less than 30%, less than 20%, or less than 10% of the therapeutic agent is released from the composition 168 hours after administration of the composition. In some embodiments, the composition is characterized in that, when administered to a subject, less than 100%, less than 90%, less than 80%, less than 70%, less than 60%, less than 50%, less than 40%, less than 30%, less than 20%, or less than 10% of the therapeutic agent is released from the composition 144 hours after administration of the composition.
  • the composition is characterized in that, when administered to a subject, less than 100%, less than 90%, less than 80%, less than 70%, less than 60%, less than 50%, less than 40%, less than 30%, less than 20%, or less than 10% of the therapeutic agent is released from the composition 6 hours after administration of the composition. In some embodiments, the composition is characterized in that, when administered to a subject, less than 100%, less than 90%, less than 80%, less than 70%, less than 60%, less than 50%, less than 40%, less than 30%, less than 20%, or less than 10% of the therapeutic agent is released from the composition 3 hours after administration of the composition.
  • the composition is characterized in that, when administered to a subject, duration of a therapeutic effect is extended by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, or at least 200% compared to duration of the therapeutic effect upon administration of the therapeutic agent alone or in a composition that is not a composition of the present disclosure.
  • the therapeutic effect is nerve block and/or reduced pain.
  • the therapeutic effect is nerve block.
  • the therapeutic effect is sciatic nerve block.
  • the therapeutic effect is reduced pain.
  • the therapeutic effect is treatment of a cancer (e.g., reduction of the size of a tumor).
  • the therapeutic effect is reduction of the size of a tumor. In some embodiments, the therapeutic effect is treatment of an allergy.
  • the composition exhibits a honeycomb-like structure with pores having a diameter of between about 1 ⁇ m and about 25 ⁇ m in size, between about 1 ⁇ m and about 20 ⁇ m in size, between about 1 ⁇ m and about 15 ⁇ m in size, between about 1 ⁇ m and about 14 ⁇ m in size, between about 1 ⁇ m and about 13 ⁇ m in size, between about 1 ⁇ m and about 12.5 ⁇ m in size, between about 1 ⁇ m and about 12 ⁇ m in size, between about 1 ⁇ m and about 11 ⁇ m in size, between about 1 ⁇ m and about 10 ⁇ m in size, between about 2.5 ⁇ m and about 25 ⁇ m in size, between about 2.5 ⁇ m and about 20 ⁇ m in size, between about 2.5 ⁇ m and about 15 ⁇ m in size, between about 2.5 ⁇ m and about 14 ⁇ m in size, between about 2.5 ⁇ m and about 25 ⁇
  • the #13447332v2 composition exhibits a honeycomb-like structure with pores having a diameter of between about 1 ⁇ m and about 25 ⁇ m in size. In some embodiments, the composition exhibits a honeycomb- like structure with pores having a diameter of between about 1 ⁇ m and about 15 ⁇ m in size. In some embodiments, the composition exhibits a honeycomb-like structure with pores having a diameter of between about 2.5 ⁇ m and about 12.5 ⁇ m in size. In some embodiments, the composition exhibits a honeycomb-like structure with pores having a diameter of between about 5 ⁇ m and about 10 ⁇ m in size. In some embodiments, the honeycomb-like structure comprises hexagonal pores.
  • the honeycomb-like structure comprises a hexagonal cavity.
  • the composition comprises a storage modulus (G′) that is greater than a loss modulus (G′′) at a temperature of about 0 °C, about 5 °C, about 10 °C, about 15 °C, about 16 °C, about 17 °C, about 18 °C, about 19 °C, about 20 °C, about 21 °C, about 22 °C, about 23 °C, about 24 °C, about 25 °C, about 26 °C, about 27 °C, about 28 °C, about 29 °C, about 30 °C, about 31 °C, about 32 °C, about 33 °C, about 34 °C, about 35 °C, about 36 °C, about 37 °C, about 38 °C, about 39 °C, about 40 °C, about 45 °C, or about 50 °C.
  • the composition comprises a storage modulus (G′) that is greater than a loss modulus (G′′) at a temperature of about 25 °C.
  • the composition is injectable.
  • the composition is biodegradable.
  • the therapeutic agent is bupivacaine present in the composition in a concentration of between about 10 mg/mL and about 50 mg/mL
  • the bovine serum albumin is present in a concentration of between about 25 mg/mL and about 125 mg/mL
  • the glutaraldehyde is present in a concentration of between about 1 mg/mL and about 15 mg/mL.
  • the therapeutic agent is bupivacaine present in the composition in a concentration of between about 10 mg/mL and about 50 mg/mL
  • the bovine serum albumin is present in a concentration of between about 37.5 mg/mL and about 112.5 mg/mL
  • the glutaraldehyde is present in a concentration of between about 2.5 mg/mL and about 12.5 mg/mL.
  • the therapeutic agent is bupivacaine present in the composition in a concentration of between about 10 mg/mL and about 50 mg/mL
  • the bovine serum albumin is present in a concentration of between about 45 mg/mL and about 105 mg/mL
  • the glutaraldehyde is present in a concentration of between about 4.5 mg/mL and about 10.5 mg/mL.
  • the therapeutic agent is bupivacaine present in the composition in a concentration of between about 10 mg/mL and about 50 mg/mL, the bovine serum albumin is present in a concentration of between about 50 mg/mL and about 100 mg/mL, and the glutaraldehyde is present in a concentration of between about 5 mg/mL and about 10 mg/mL.
  • the therapeutic agent is bupivacaine present in the composition in a concentration of about 20 mg/mL
  • the bovine serum albumin is present in a concentration of about 50 mg/mL
  • the glutaraldehyde is present in a concentration of about 5 mg/mL.
  • the therapeutic agent is bupivacaine present in the composition in a concentration of about 20 mg/mL, the bovine serum albumin is present in a concentration of about 50 mg/mL, and the glutaraldehyde is present in a concentration of about 10 mg/mL. In some embodiments, the therapeutic agent is bupivacaine present in the composition in a concentration of about 20 mg/mL, the bovine serum albumin is present in a concentration of about 100 mg/mL, and the glutaraldehyde is present in a concentration of about 5 mg/mL.
  • the therapeutic agent is bupivacaine present in the composition in a concentration of about 20 mg/mL, the bovine serum albumin is present in a concentration of about 100 mg/mL, and the glutaraldehyde is present in a concentration of about 10 mg/mL.
  • the therapeutic agent is an anti-cancer agent, the crosslinking agent is of Formula (II), and the protein is bovine serum albumin or alpha-1-acid glycoprotein.
  • the therapeutic agent is an anti-cancer agent, the crosslinking agent is of Formula (II), and the protein is bovine serum albumin.
  • the therapeutic agent is an anti-cancer agent, the crosslinking agent is of Formula (II), and the protein is alpha- 1-acid glycoprotein.
  • the therapeutic agent is an anti-cancer agent, the crosslinking agent is glutaraldehyde, and the protein is bovine serum albumin or alpha-1-acid glycoprotein.
  • the therapeutic agent is an anti-cancer agent, the crosslinking agent is glutaraldehyde, and the protein is bovine serum albumin.
  • the therapeutic agent is an anti-cancer agent, the crosslinking agent is glutaraldehyde, and the protein is alpha-1-acid glycoprotein.
  • the therapeutic agent is a free base form of an anti-cancer agent, the crosslinking agent is of Formula (II), and the protein is bovine serum albumin or alpha-1- acid glycoprotein.
  • the therapeutic agent is a free base form of an anti- cancer agent, the crosslinking agent is of Formula (II), and the protein is bovine serum albumin.
  • the therapeutic agent is a free base form of an anti-cancer agent, the crosslinking agent is of Formula (II), and the protein is alpha-1-acid glycoprotein.
  • the therapeutic agent is a free base form of an anti-cancer agent
  • the crosslinking agent is glutaraldehyde
  • the protein is bovine serum albumin or alpha-1-acid glycoprotein.
  • the therapeutic agent is a free base form of an anti-cancer agent
  • the crosslinking agent is glutaraldehyde
  • the protein is bovine serum albumin.
  • the therapeutic agent is a free base form of an anti-cancer agent
  • the crosslinking agent is glutaraldehyde
  • the protein is alpha-1-acid glycoprotein.
  • the therapeutic agent is paclitaxel, the crosslinking agent is of Formula (II), and the protein is bovine serum albumin or alpha-1-acid glycoprotein.
  • the therapeutic agent is paclitaxel, the crosslinking agent is of Formula (II), and the protein is bovine serum albumin.
  • the therapeutic agent is paclitaxel, the crosslinking agent is of Formula (II), and the protein is alpha-1-acid glycoprotein.
  • the therapeutic agent is paclitaxel, the crosslinking agent is glutaraldehyde, and the protein is bovine serum albumin or alpha-1-acid glycoprotein.
  • the therapeutic agent is paclitaxel
  • the crosslinking agent is glutaraldehyde
  • the protein is bovine serum albumin.
  • the therapeutic agent is paclitaxel
  • the crosslinking agent is glutaraldehyde
  • the protein is alpha-1-acid glycoprotein.
  • the therapeutic agent is paclitaxel present in the composition in a concentration of between about 0.1 mg/mL and about 10 mg/mL
  • the bovine serum albumin is present in a concentration of between about 25 mg/mL and about 125 mg/mL
  • the glutaraldehyde is present in a concentration of between about 1 mg/mL and about 15 mg/mL.
  • the therapeutic agent is paclitaxel present in the composition in a concentration of between about 0.1 mg/mL and about 10 mg/mL
  • the bovine serum albumin is present in a concentration of between about 45 mg/mL and about 105 mg/mL
  • the glutaraldehyde is present in a concentration of between about 4.5 mg/mL and about 10.5 mg/mL.
  • the therapeutic agent is paclitaxel present in the composition in a concentration of between about 0.1 mg/mL and about 10 mg/mL, the bovine serum albumin is present in a concentration of between about 50 mg/mL and about 100 mg/mL, and the glutaraldehyde is present in a concentration of between about 5 mg/mL and about 10 mg/mL.
  • the therapeutic agent is paclitaxel present in the composition in a concentration of about 1 mg/mL
  • the bovine serum albumin is present in a concentration of about 50 mg/mL
  • the glutaraldehyde is present in a concentration of about 5 mg/mL.
  • the therapeutic agent is paclitaxel present in the composition in a concentration of about 1 mg/mL, the bovine serum albumin is present in a concentration of about 50 mg/mL, and the glutaraldehyde is present in a concentration of about 10 mg/mL. In some embodiments, the therapeutic agent is paclitaxel present in the composition in a concentration of about 1 mg/mL, the bovine serum albumin is present in a concentration of about 100 mg/mL, and the glutaraldehyde is present in a concentration of about 5 mg/mL.
  • the therapeutic agent is paclitaxel present in the composition in a concentration of about 1 mg/mL, the bovine serum albumin is present in a concentration of about 100 mg/mL, and the glutaraldehyde is present in a concentration of about 10 mg/mL.
  • the therapeutic agent is an antihistamine
  • the crosslinking agent is of Formula (II)
  • the protein is bovine serum albumin or alpha-1-acid glycoprotein.
  • the therapeutic agent is an antihistamine
  • the crosslinking agent is of Formula (II)
  • the protein is bovine serum albumin.
  • the therapeutic agent is an antihistamine, the crosslinking agent is of Formula (II), and the protein is alpha-1-acid glycoprotein.
  • the therapeutic agent is an antihistamine, the crosslinking agent is glutaraldehyde, and the protein is bovine serum albumin or alpha-1-acid glycoprotein.
  • the therapeutic agent is an antihistamine, the crosslinking agent is glutaraldehyde, and the protein is bovine serum albumin.
  • the therapeutic agent is an antihistamine, the crosslinking agent is glutaraldehyde, and the protein is alpha-1- acid glycoprotein.
  • the therapeutic agent is a free base form of an antihistamine, the crosslinking agent is of Formula (II), and the protein is bovine serum albumin or alpha-1-acid glycoprotein.
  • the therapeutic agent is a free base form of an antihistamine, the crosslinking agent is of Formula (II), and the protein is bovine serum albumin.
  • the therapeutic agent is a free base form of an antihistamine, the crosslinking agent is of Formula (II), and the protein is alpha-1-acid glycoprotein.
  • the therapeutic agent is a free base form of an antihistamine
  • the crosslinking agent is glutaraldehyde
  • the protein is bovine serum albumin or alpha-1-acid glycoprotein.
  • the therapeutic agent is a free base form of an antihistamine
  • the crosslinking agent is glutaraldehyde
  • the protein is bovine serum albumin.
  • the therapeutic agent is a free base form of an antihistamine
  • the crosslinking agent is glutaraldehyde
  • the protein is alpha-1-acid glycoprotein.
  • the therapeutic agent is diphenydramine, the crosslinking agent is of Formula (II), and the protein is bovine serum albumin or alpha-1-acid glycoprotein.
  • the therapeutic agent is diphenydramine, the crosslinking agent is of Formula (II), and the protein is bovine serum albumin.
  • the therapeutic agent is diphenydramine, the crosslinking agent is of Formula (II), and the protein is alpha-1-acid glycoprotein.
  • the therapeutic agent is diphenydramine, the crosslinking agent is glutaraldehyde, and the protein is bovine serum albumin or alpha-1-acid glycoprotein.
  • the therapeutic agent is diphenydramine, the crosslinking agent is glutaraldehyde, and the protein is bovine serum albumin.
  • the therapeutic #13447332v2 agent is diphenydramine, the crosslinking agent is glutaraldehyde, and the protein is alpha-1- acid glycoprotein.
  • the therapeutic agent is a free base form of diphenydramine, the crosslinking agent is of Formula (II), and the protein is bovine serum albumin or alpha-1-acid glycoprotein.
  • the therapeutic agent is a free base form of diphenydramine, the crosslinking agent is of Formula (II), and the protein is bovine serum albumin.
  • the therapeutic agent is a free base form of diphenydramine, the crosslinking agent is of Formula (II), and the protein is alpha-1-acid glycoprotein.
  • the therapeutic agent is a free base form of diphenydramine, the crosslinking agent is glutaraldehyde, and the protein is bovine serum albumin or alpha-1-acid glycoprotein.
  • the therapeutic agent is a free base form of diphenydramine, the crosslinking agent is glutaraldehyde, and the protein is bovine serum albumin.
  • the therapeutic agent is a free base form of diphenydramine
  • the crosslinking agent is glutaraldehyde
  • the protein is alpha-1-acid glycoprotein.
  • the therapeutic agent is diphenydramine present in the composition in a concentration of between about 0.1 mg/mL and about 10 mg/mL
  • the bovine serum albumin is present in a concentration of between about 25 mg/mL and about 125 mg/mL
  • the glutaraldehyde is present in a concentration of between about 1 mg/mL and about 15 mg/mL.
  • the therapeutic agent is diphenydramine present in the composition in a concentration of between about 0.1 mg/mL and about 10 mg/mL
  • the bovine serum albumin is present in a concentration of between about 45 mg/mL and about 105 mg/mL
  • the glutaraldehyde is present in a concentration of between about 4.5 mg/mL and about 10.5 mg/mL.
  • the therapeutic agent is diphenydramine present in the composition in a concentration of between about 0.1 mg/mL and about 10 mg/mL, the bovine serum albumin is present in a concentration of between about 50 mg/mL and about 100 mg/mL, and the glutaraldehyde is present in a concentration of between about 5 mg/mL and about 10 mg/mL.
  • the therapeutic agent is diphenydramine present in the composition in a concentration of about 1 mg/mL, the bovine serum albumin is present in a concentration of about 50 mg/mL, and the glutaraldehyde is present in a concentration of about 5 mg/mL.
  • the therapeutic agent is diphenydramine present in the #13447332v2 composition in a concentration of about 1 mg/mL, the bovine serum albumin is present in a concentration of about 50 mg/mL, and the glutaraldehyde is present in a concentration of about 10 mg/mL. In some embodiments, the therapeutic agent is diphenydramine present in the composition in a concentration of about 1 mg/mL, the bovine serum albumin is present in a concentration of about 100 mg/mL, and the glutaraldehyde is present in a concentration of about 5 mg/mL.
  • the preparation further comprises adding 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.
  • 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 #13447332v2 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, for example, one-half or one-third of such a dosage.
  • 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, 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, mucosal, nasal, bucal, sublingual
  • intratracheal instillation, bronchial instillation, and/or inhalation and/or as an oral spray, nasal spray, and/
  • 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 in a subject in need thereof, in preventing a disease in a subject in need thereof, and/or in reducing the risk to develop a disease in a subject in need thereof), improve bioavailability, improve their ability to cross the blood-brain barrier, improve safety, reduce drug resistance, reduce and/or modify metabolism, inhibit excretion, and/or modify distribution in a subject or cell.
  • additional pharmaceutical agents that improve their activity (e.g., activity (e.g., potency and/or efficacy) in treating a disease in a subject in need thereof, in preventing a disease in a subject in need thereof, and/or in reducing the risk to develop a disease in a subject in need thereof), improve bioavailability, improve their ability to cross the blood-brain barrier, improve safety, reduce drug resistance, reduce and/or modify metabolism, inhibit excretion, and/or modify
  • a pharmaceutical composition described herein including a therapeutic agent described herein and an additional agent exhibits a synergistic effect that is absent in a pharmaceutical composition including one of the therapeutic agent and the additional agent, but not both.
  • the additional agent is a therapeutic agent.
  • the 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
  • 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.
  • 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 #13447332v2 herein in a single dose or administered separately in different doses.
  • 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.
  • the additional pharmaceutical agent(s) in combination be utilized at levels that do not exceed the levels at which they are utilized individually.
  • the levels utilized in combination will be lower than those utilized individually.
  • the subject is an animal.
  • the animal may be of either sex and may be at any stage of development.
  • 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
  • kits e.g., pharmaceutical packs.
  • the kits provided comprise a composition described herein and instructions for using the kit.
  • the kits provided may comprise a composition described herein and 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 composition described herein.
  • kits including a first container comprising a composition described herein.
  • the kits are useful for treating a disease or condition (e.g., pain) in a subject in need thereof.
  • the kits are useful for preventing a disease or condition (e.g., pain) in a subject in need thereof.
  • the kits are useful for reducing the risk of developing a disease or condition (e.g., pain) in a subject in need thereof.
  • a kit described herein may also include information as required by a regulatory agency such as the U.S. Food and Drug Administration (FDA).
  • FDA U.S. Food and Drug Administration
  • kits described herein may include one or more additional pharmaceutical agents described herein as a separate composition.
  • Methods of Use The present disclosure also provides methods for treating diseases or conditions in a subject in need thereof, the methods comprising administering to the subject a provided composition.
  • the present disclosure also provides methods for preventing diseases or conditions in a subject in need thereof, the methods comprising administering to the subject a provided composition.
  • the present disclosure also provides methods for reducing the risk of developing a disease or condition in a subject in need thereof.
  • the disclosed compositions can be used to treat a subject (e.g., a human) having a disease or condition that would benefit from administration of a therapeutic agent.
  • the subject is administered an effective amount of any one or more disclosed compositions.
  • the subject is administered a therapeutically effective amount of any one or more disclosed compositions.
  • the subject is administered a prophylactically effective amount of any one or more disclosed compositions.
  • the subject is an animal.
  • the subject is a mammal.
  • the subject is a human.
  • the subject is a human aged 18 years or older.
  • the subject is a human aged 12-18 years, exclusive.
  • the subject is a human aged 2-12 years, inclusive. In some embodiments, the subject is a human younger than 2 years. In some embodiments, the subject is a non-human animal. In some embodiments, the subject is a non-human mammal. In some embodiments, the disease or condition being treated is one associated with a specific biological target. In some embodiments, the target is a voltage-gated sodium channel. In some embodiments, the target is a site-1 sodium channel. In some embodiments, the disease or condition is pain. The quality of life of patients suffering from postoperative or even chronic pain is often diminished by the need for repeated administration of systemic analgesic medications (e.g., opioids), which give rise to potentially serious complications and clouding of the sensorium.
  • systemic analgesic medications e.g., opioids
  • tetrodotoxin is an attractive candidate in peripheral nerve anesthesia because of its reduced affinity for voltage dependent sodium channels of the peripheral nerve, but a poor affinity for the cardiac sodium channel isoform. It also does not cross the blood brain barrier.
  • Voltage-gated sodium channels play important roles in nociceptive nerve conduction, but candidate anesthetics (e.g., specific antagonists of sodium channels) are often not effective in vivo because of lack of permeability of the perineurial barrier.
  • candidate anesthetics e.g., specific antagonists of sodium channels
  • permeation enhancers have been used to increase the permeability of lipid barriers and, they can be associated with myotoxicity.
  • the methods for treating diseases or conditions comprise administering the composition such that the therapeutic agent is delivered to a biological target at a rate that is less than if the therapeutic agent were administered alone.
  • the therapeutic agent is delivered via sustained release.
  • the administration is local such that the sustained release is a local sustained release (i.e., local administration for a local effect, e.g., subcutaneous administration).
  • administration is systemic such that the sustained release is a systemic sustained release (i.e., systemic administration for a dispersed or system-wide effect, e.g., parenteral administration).
  • a systemic sustained release i.e., systemic administration for a dispersed or system-wide effect, e.g., parenteral administration.
  • less than 100%, less than 90%, less than 80%, less than 70%, less than 60%, less than 50%, less than 40%, less than 30%, less than 20%, or less than 10% of the therapeutic agent is released from the composition 168 hours after administration of the composition to the subject.
  • less than 100%, less than 90%, less than 80%, less than 70%, less than 60%, less than 50%, less than 40%, less than 30%, less than 20%, or less than 10% of the therapeutic agent is released from the composition 144 hours after administration of the composition to the subject.
  • less than 100%, less than 90%, less than 80%, less than 70%, less than 60%, less than 50%, less than 40%, less than 30%, less than 20%, or less than 10% of the therapeutic agent is released from the composition 120 hours after administration of the composition to the subject. In some embodiments, less than 100%, less than 90%, less than 80%, less than 70%, less than 60%, less than 50%, less than 40%, less than 30%, less than 20%, or less than 10% of the therapeutic agent is released from the composition 96 hours after administration of the composition to the subject.
  • less than 100%, less than 90%, less than 80%, less than 70%, less than 60%, less than 50%, less than 40%, less than 30%, less than 20%, or less than 10% of the therapeutic agent is released from the composition 72 hours after administration of the composition to the subject. In some embodiments, less than 100%, less than 90%, less than 80%, less than 70%, less than 60%, less than 50%, less than 40%, less than 30%, less than 20%, or less than 10% of the therapeutic agent is released from the composition 48 hours after administration of the composition to the subject.
  • less than 100%, less than 90%, less than 80%, less than 70%, less than 60%, less than 50%, less than 40%, less than 30%, less than 20%, or less than 10% of the therapeutic agent is released from the composition 24 hours after administration of the composition to the subject. In some embodiments, less than 100%, less than 90%, less than 80%, less than 70%, less than 60%, less than 50%, less than 40%, less than 30%, less than 20%, or less than 10% of the therapeutic agent is released from the composition 12 hours after administration of the composition to the subject.
  • less than 100%, less than 90%, less than 80%, less than 70%, less than 60%, less than 50%, less than 40%, less than 30%, less than 20%, or less than 10% of the therapeutic agent is released from the composition 6 hours after administration of the composition to the subject. In some embodiments, less than 100%, less than 90%, less than 80%, less than 70%, less than 60%, less than 50%, less than 40%, less than 30%, less than 20%, or less than 10% of the therapeutic agent is released from the composition 3 hours after administration of the composition to the subject.
  • duration of a therapeutic effect is extended by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 200%, at least 300%, at least 400%, at least 500%, at least 600%, at least 700%, at least 800%, at least 900%, at least 1,000%, or at least 10,000% compared to duration of a therapeutic effect upon administration of the therapeutic agent alone or in a composition that is not a provided composition.
  • duration of a therapeutic effect is extended by at least 10% compared to duration of a therapeutic effect upon administration of the therapeutic agent alone or in a composition that is not a provided composition.
  • duration of a therapeutic effect is extended by at least 20% compared to duration of a therapeutic effect upon administration of the therapeutic agent alone or in a composition that is not a provided composition. In some embodiments, duration of a therapeutic effect is extended by at least 30% compared to duration of a therapeutic effect upon administration of the therapeutic agent alone or in a composition that is not a provided composition. In some embodiments, duration of a therapeutic effect is extended by at least 40% compared to duration of a therapeutic effect upon administration of the therapeutic agent alone or in a composition that is not a provided composition. In some embodiments, duration of a #13447332v2 therapeutic effect is extended by at least 50% compared to duration of a therapeutic effect upon administration of the therapeutic agent alone or in a composition that is not a provided composition.
  • duration of a therapeutic effect is extended by at least 60% compared to duration of a therapeutic effect upon administration of the therapeutic agent alone or in a composition that is not a provided composition. In some embodiments, duration of a therapeutic effect is extended by at least 70% compared to duration of a therapeutic effect upon administration of the therapeutic agent alone or in a composition that is not a provided composition. In some embodiments, duration of a therapeutic effect is extended by at least 80% compared to duration of a therapeutic effect upon administration of the therapeutic agent alone or in a composition that is not a provided composition. In some embodiments, duration of a therapeutic effect is extended by at least 90% compared to duration of a therapeutic effect upon administration of the therapeutic agent alone or in a composition that is not a provided composition.
  • duration of a therapeutic effect is extended by at least 100% compared to duration of a therapeutic effect upon administration of the therapeutic agent alone or in a composition that is not a provided composition. In some embodiments, duration of a therapeutic effect is extended by least 200% compared to duration of a therapeutic effect upon administration of the therapeutic agent alone or in a composition that is not a provided composition. In some embodiments, duration of a therapeutic effect is extended by least 300% compared to duration of a therapeutic effect upon administration of the therapeutic agent alone or in a composition that is not a provided composition. In some embodiments, duration of a therapeutic effect is extended by least 400% compared to duration of a therapeutic effect upon administration of the therapeutic agent alone or in a composition that is not a provided composition.
  • duration of a therapeutic effect is extended by least 500% compared to duration of a therapeutic effect upon administration of the therapeutic agent alone or in a composition that is not a provided composition. In some embodiments, duration of a therapeutic effect is extended by least 600% compared to duration of a therapeutic effect upon administration of the therapeutic agent alone or in a composition that is not a provided composition. In some embodiments, duration of a therapeutic effect is extended by least 700% compared to duration of a therapeutic effect upon administration of the therapeutic agent alone or in a composition that is not a provided composition. In some embodiments, duration of a therapeutic effect is extended by least 800% compared to duration of a therapeutic effect upon administration of the therapeutic agent alone or in a composition that is not a provided composition.
  • duration of a therapeutic effect is extended by least 900% compared to duration of a therapeutic effect upon administration of the therapeutic agent alone or in a composition that is not a provided composition. In some embodiments, duration of a #13447332v2 therapeutic effect is extended by least 1,000% compared to duration of a therapeutic effect upon administration of the therapeutic agent alone or in a composition that is not a provided composition. In some embodiments, duration of a therapeutic effect is extended by least 10,000% compared to duration of a therapeutic effect upon administration of the therapeutic agent alone or in a composition that is not a provided composition. In some embodiments, the therapeutic effect is nerve block and/or reduced pain. In some embodiments, the therapeutic effect is nerve block or reduced pain. In some embodiments, the therapeutic effect is nerve block and reduced pain.
  • the therapeutic effect is nerve block. In some embodiments, the therapeutic effect is sciatic nerve block. In some embodiments, the therapeutic effect is reduced pain. In some embodiments, the therapeutic effect is reduction of pain by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 99% compared to a baseline level of pain prior to administration.
  • the present disclosure also provides uses of a provided composition in a method described herein. The present disclosure also provides uses of a provided pharmaceutical composition in a method described herein. The present disclosure also provides a provided composition for use in a method described herein. The present disclosure also provides a provided pharmaceutical composition for use in a method described herein.
  • the present disclosure also provides methods for preparing a provided composition, wherein the method comprises: providing a therapeutic agent or a diagnostic agent; forming a protein-stabilized nanoparticle comprising the therapeutic agent or the diagnostic agent by exposing the therapeutic agent or the diagnostic agent to a solution of a protein in water; and reacting the protein with a crosslinking agent, thereby producing a crosslinked protein matrix.
  • providing the therapeutic agent or the diagnostic agent further comprises forming a free base form of the therapeutic agent or the diagnostic agent.
  • the free base form of the therapeutic agent or the diagnostic agent is formed by reacting a salt form of the therapeutic agent or the diagnostic agent with a base.
  • the method comprises providing a diagnostic agent.
  • providing the diagnostic agent further comprises forming a free base form of the #13447332v2 diagnostic agent.
  • the free base form of the diagnostic agent is formed by reacting a salt form of the diagnostic agent with a base.
  • the method comprises providing a therapeutic agent.
  • providing the therapeutic agent further comprises forming a free base form of the therapeutic agent.
  • the free base form of the therapeutic agent is formed by reacting a salt form of the therapeutic agent with a base.
  • the therapeutic agent is a small molecule or a biologic.
  • the therapeutic agent is a small molecule, a protein, or a nucleic acid.
  • the therapeutic agent is a small molecule.
  • the therapeutic agent is an anesthetic. In some embodiments, the therapeutic agent is a free base form of an anesthetic. In some embodiments, the therapeutic agent is hydrophobic. In some embodiments, the therapeutic agent is hydrohilic. In some embodiments, the therapeutic agent does not comprise a primary or secondary amine. In some embodiments, the therapeutic agent is not doxorubicin.
  • the therapeutic agent is a local anesthetic (e.g., tetrodotoxin, saxitoxin, neosaxitoxin, bupivacaine, amylocaine, ambucaine, articaine, benzocaine, benzonatate, butacaine, butanilicaine, carbocaine, cepastat, chloraseptic, chloroprocaine, cinchocaine, citanest, cyclomethycaine, dibucaine, diperodon, dimethocaine, eucaine, etidocaine, fomocaine, fotocaine, hydroxyprocaine, isobucaine, levobupivacaine, lidocaine, marcaine, mepivacaine, meprylcaine, metabutoxycaine, nitracaine, orthocaine, orabloc, oxetacaine, oxybuprocaine, paraethoxycaine, phenacaine
  • the therapeutic agent is a free base form of a local anesthetic.
  • the therapeutic agent is a local anesthetic compound, for example, but not limited to, an amino ester compound (e.g., procaine, tetracaine, chloroprocaine, benzocaine, butacaine, dimethocaine) or an amino amide compound (e.g., procainamide, lidocaine).
  • the local anesthetic is a sodium channel blocker, for example, a site 1 sodium channel blocker (e.g., tetrodotoxin, saxitoxins (saxitoxin, neosaxitoxin), gonyautoxins (gonyautoxin V, gonyautoxin VI), ⁇ - conotoxins)) or an amino amide local anesthetic.
  • the therapeutic agent is a site 1 sodium channel blocker, amino ester anesthetic, or an amino amide anesthetic, or a derivative thereof.
  • the therapeutic agent is tetrodotoxin, saxitoxin, neosaxitoxin, bupivacaine, amylocaine, ambucaine, articaine, benzocaine, benzonatate, butacaine, #13447332v2 butanilicaine, carbocaine, cepastat, chloraseptic, chloroprocaine, cinchocaine, citanest, cyclomethycaine, dibucaine, diperodon, dimethocaine, eucaine, etidocaine, fomocaine, fotocaine, hydroxyprocaine, isobucaine, levobupivacaine, lidocaine, marcaine, mepivacaine, meprylcaine, metabutoxycaine, nitracaine, orthocaine, orabloc, oxetacaine, oxybuprocaine, paraethoxycaine, phenacaine, piperocaine, orabloc,
  • the therapeutic agent is bupivacaine. In some embodiments, the therapeutic agent is a free base form of bupivacaine.
  • the present disclosure also provides methods for preparing a provided composition, wherein the method comprises: providing an anesthetic; forming a nanoparticle comprising the anesthetic by exposing the anesthetic to a solution of water comprising a protein; and reacting the protein with a crosslinking agent, thereby producing a crosslinked protein matrix.
  • providing the anesthetic further comprises forming a free base form of the anesthetic.
  • the free base form of the anesthetic is formed by reacting a salt form of the anesthetic with a base.
  • the protein is not treated with ethanol prior to reacting with the crosslinking agent. In some embodiments, the protein is not denatured prior to reacting with the crosslinking agent. In some embodiments, the protein is bovine serum albumin or alpha-1-acid glycoprotein. In some embodiments, the protein is bovine serum albumin. In some embodiments, the protein is alpha-1-acid glycoprotein. In some embodiments, the crosslinking agent is of Formula (II): (II), wherein L is optionally substituted alkylene or optionally substituted heteroalkylene. In some embodiments, L is optionally substituted alkylene. In some embodiments, L is optionally substituted C 1-12 alkylene.
  • L is optionally substituted C 1-6 alkylene. In some embodiments, L is optionally substituted C1-3 alkylene. #13447332v2 In some embodiments, L is unsubstituted alkylene. In some embodiments, L is unsubstituted C 1-12 alkylene. In some embodiments, L is unsubstituted C 1-6 alkylene. In some embodiments, L is unsubstituted C 1-3 alkylene.
  • L is unsubstituted C 12 alkylene, unsubstituted C11 alkylene, unsubstituted C10 alkylene, unsubstituted C9 alkylene, unsubstituted C8 alkylene, unsubstituted C7 alkylene, unsubstituted C6 alkylene, unsubstituted C5 alkylene, unsubstituted C 4 alkylene, unsubstituted C 3 alkylene, unsubstituted C 2 alkylene, or unsubstituted C1 alkylene.
  • L is unsubstituted C12 alkylene.
  • L is unsubstituted C11 alkylene.
  • L is unsubstituted C10 alkylene. In some embodiments, L is unsubstituted C 9 alkylene. In some embodiments, L is unsubstituted C8 alkylene. In some embodiments, L is unsubstituted C7 alkylene. In some embodiments, L is unsubstituted C6 alkylene. In some embodiments, L is unsubstituted C5 alkylene. In some embodiments, L is unsubstituted C 4 alkylene. In some embodiments, L is unsubstituted C 3 alkylene. In some embodiments, L is unsubstituted C 2 alkylene. In some embodiments, L is unsubstituted C1 alkylene.
  • L is methylene, ethylene, n-propylene, isopropylene, n-butylene, tert-butylene, sec-butylene, isobutylene, n-pentylene, 3- pentanylene, amylene, neopentylene, 3-methylene-2-butanylene, tert-amylene, or n-hexylene.
  • L is methylene, ethylene, n-propylene, n-butylene, n-pentylene, or n- hexylene.
  • L is methylene.
  • L is ethylene.
  • L is n-propylene.
  • L is n-butylene. In some embodiments, L is n-pentylene. In some embodiments, L is n-hexylene.
  • the crosslinking agent is glutaraldehyde. In some embodiments, the crosslinking agent is of Formula (II), and the protein is bovine serum albumin or alpha-1-acid glycoprotein. In some embodiments, the crosslinking agent is of Formula (II), and the protein is bovine serum albumin. In some embodiments, the crosslinking agent is of Formula (II), and the protein is alpha-1-acid glycoprotein.
  • the crosslinking agent is glutaraldehyde, and the protein is bovine serum albumin or alpha-1-acid glycoprotein. In some embodiments, the crosslinking agent is glutaraldehyde, and the protein is bovine serum albumin. In some embodiments, the crosslinking agent is glutaraldehyde, and the protein is alpha-1-acid glycoprotein. In some embodiments, the therapeutic agent is an anesthetic, the crosslinking agent is of Formula (II), and the protein is bovine serum albumin or alpha-1-acid glycoprotein. In some embodiments, the therapeutic agent is an anesthetic, the crosslinking agent is of Formula (II), and the protein is bovine serum albumin.
  • the therapeutic agent is an anesthetic, the crosslinking agent is of Formula (II), and the protein is alpha-1-acid glycoprotein.
  • the therapeutic agent is an anesthetic, the crosslinking agent is #13447332v2 glutaraldehyde, and the protein is bovine serum albumin or alpha-1-acid glycoprotein.
  • the therapeutic agent is an anesthetic, the crosslinking agent is glutaraldehyde, and the protein is bovine serum albumin.
  • the therapeutic agent is an anesthetic, the crosslinking agent is glutaraldehyde, and the protein is alpha-1-acid glycoprotein.
  • the therapeutic agent is a free base form of an anesthetic, the crosslinking agent is of Formula (II), and the protein is bovine serum albumin or alpha-1-acid glycoprotein.
  • the therapeutic agent is a free base form of an anesthetic, the crosslinking agent is of Formula (II), and the protein is bovine serum albumin.
  • the therapeutic agent is a free base form of an anesthetic, the crosslinking agent is of Formula (II), and the protein is alpha-1-acid glycoprotein.
  • the therapeutic agent is a free base form of an anesthetic
  • the crosslinking agent is glutaraldehyde
  • the protein is bovine serum albumin or alpha-1-acid glycoprotein.
  • the therapeutic agent is a free base form of an anesthetic
  • the crosslinking agent is glutaraldehyde
  • the protein is bovine serum albumin.
  • the therapeutic agent is a free base form of an anesthetic
  • the crosslinking agent is glutaraldehyde
  • the protein is alpha-1- acid glycoprotein.
  • the therapeutic agent is bupivacaine, the crosslinking agent is of Formula (II), and the protein is bovine serum albumin or alpha-1-acid glycoprotein.
  • the therapeutic agent is bupivacaine, the crosslinking agent is of Formula (II), and the protein is bovine serum albumin.
  • the therapeutic agent is bupivacaine, the crosslinking agent is of Formula (II), and the protein is alpha-1-acid glycoprotein.
  • the therapeutic agent is bupivacaine, the crosslinking agent is glutaraldehyde, and the protein is bovine serum albumin or alpha-1-acid glycoprotein.
  • the therapeutic agent is bupivacaine, the crosslinking agent is glutaraldehyde, and the protein is bovine serum albumin. In some embodiments, the therapeutic agent is bupivacaine, the crosslinking agent is glutaraldehyde, and the protein is alpha-1-acid glycoprotein. In some embodiments, the therapeutic agent is a free base form of bupivacaine, the crosslinking agent is of Formula (II), and the protein is bovine serum albumin or alpha-1-acid glycoprotein. In some embodiments, the therapeutic agent is a free base form of bupivacaine, the crosslinking agent is of Formula (II), and the protein is bovine serum albumin.
  • the therapeutic agent is a free base form of bupivacaine, the crosslinking agent is of Formula (II), and the protein is alpha-1-acid glycoprotein.
  • the therapeutic agent is a free base form of bupivacaine, the crosslinking agent is glutaraldehyde, and the protein is bovine serum albumin or alpha-1-acid glycoprotein.
  • the #13447332v2 therapeutic agent is a free base form of bupivacaine, the crosslinking agent is glutaraldehyde, and the protein is bovine serum albumin.
  • 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.
  • the invention 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 claim 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.
  • BNP@BSA-X A glutaraldehyde-crosslinked bovine serum albumin hydrogel (BSA-X) containing bupivacaine free base nanoparticles (BNPs), abbreviated BNP@BSA-X, was made by a one-pot approach (FIG.1). In brief (see Methods) the free base of bupivacaine was produced by alkaline precipitation of bupivacaine hydrochloride.
  • BNP bovine serum albumin
  • PBS phosphate-buffered saline
  • BNP@BSA bupivacaine nanoparticles
  • the methanol was evaporated by rotary evaporation.
  • G glutaraldehyde
  • BNP@BSA-X BSA hydrogel with embedded BNPs
  • BNP@BSA ⁇ 7 nm NPs, corresponding to BSA protein.
  • BNP@BSA ⁇ 7 nm NPs, corresponding to BSA protein.
  • BNP@BSA measured 70-100 nm by transmission electron microscopy (TEM; FIG.2B).
  • TEM transmission electron microscopy
  • #13447332v2 BNP@BSA-X was synthesized using 50 mg/ml or 100 mg/ml of BSA and 5 mg/ml or 10 mg/ml of GA.
  • all BNP@BSA-X samples formed hydrogels (FIG.8).
  • the gelation time (the time after the addition of GA for storage moduli to plateau) was shorter with 100 mg/ml BSA than 50 mg/ml BSA (FIG.9).
  • the BNP@BSA-X hydrogels had a storage modulus (G') greater than the loss modulus (G'') at all concentrations of BSA and GA, indicating solid-like properties (FIG.10).
  • the higher GA concentration (10 mg/ml) resulted in a higher G' (FIGs.2C and 10), likely due to increased crosslinking between BSA proteins. Since all the BNP@BSA-X formed hydrogels, 50 mg/ml BSA (with 5 or 10 mg/ml of GA) was used in downstream experiments, to minimize the mass to be injected in vivo.
  • the design is of drug nanoparticles contained within a cross-linked albumin hydrogel.
  • Cross-linking would occur prior to administration a) to ensure that the cross-linking occurs within the drug delivery system rather than with tissues, b) to minimize the extent to which the system will dissipate after injection, and c) to reduce the need for rapid cross-linking in vivo.
  • shear-thinning properties i.e., for the material to be highly elastic prior to injection, be readily injectable, then to regain elasticity after injection.
  • the mechanical properties of BNP@BSA-X formulations were evaluated by rheometry through step-strain oscillations (FIGs.2D and 11).
  • BNP@BSA-X (5 mg/ml GA)
  • the application of strain 500%) led to liquid-like properties, with G' ⁇ G'' (FIG.2D).
  • G' exceeded G'' within seconds.
  • BNP@BSA-X (10 mg/ml GA)
  • G' closely approached G'', suggesting a lack of shear-thinning behavior.
  • BNP@BSA with 10 mg/ml GA was difficult to inject through a 23-gauge needle.
  • Bupivacaine release from BNP@BSA and BNP@BSA-X were much slower than from BNP in PBS (both p ⁇ 0.0001 at 24 hr), underscoring the significant role of BSA in slowing release.
  • Nanoparticles were prepared from both drugs and incorporated into crosslinked BSA.
  • the free base form of diphenhydramine was generated by adding triethylamine (TEA) to diphenhydramine•HCl in methanol then added dropwise to a 50 mg/ml BSA solution in pH 7.4 PBS.
  • TAA triethylamine
  • G glutaraldehyde
  • DHNP@BSA-X diphenhydramine nanoparticles in crosslinked BSA
  • Paclitaxel was dissolved in methanol and added to a 50 mg/ml BSA solution in pH 7.4 PBS.
  • the duration of sensory block from BNP@BSA increased with increasing bupivacaine nanoparticle concentration (2.9 hr for 10 mg/ml and 3.8 hr for 20 mg/ml), but the increase was not statistically significant, and reached a maximum duration of approximately 4 hr.
  • BNP@BSA-X achieved 39.9 h of nerve block.
  • the duration achieved by the same volume of commercially available 13.3 mg/ml bupivacaine liposomes (Exparel ® ) was 5.1 h, i.e., 8-fold shorter (p ⁇ 0.0001).
  • Sensory nerve block with 20 mg/ml bupivacaine•HCl@BSA-X (7.3 hr) was 5.4-fold shorter than with BNP@BSA-X (39.9 hr, p ⁇ 0.0001). This observation aligns with in vitro release kinetics (FIG.2E).
  • the durations of nerve block and tissue retention from bupivacaine•HCl@BSA-X were much shorter than from BNP@BSA-X, supporting the benefit of nanoformulation.
  • the duration of nerve block and tissue retention from crosslinked BSA (BNP@BSA-X and bupivacaine•HCl@BSA-X) were much longer than from non-crosslinked BSA (BNP@BSA), indicating the significance of BSA cross-linking.
  • the duration of nerve block also correlated with the percentage of fluorescence relative intensity of Cy5.5 dye remaining on day 2 (FIG.6B), as calculated from data in FIG.5D.
  • cross-linking of BSA BNP@BSA-X vs. BNP@BSA was associated with much longer duration of block.
  • Tissue reaction Tissue reaction was assessed 4 days after the injection of BNP@BSA-X (20 mg/mL bupivacaine) in rat subjects, a time frame when acute inflammation and muscle injury are typically well-established following perineural injection of drugs and biomaterials (FIGs.7A- 7E).
  • the sciatic nerves and adjacent tissues were harvested, processed into hematoxylin-eosin (H&E) stained sections and inflammation and myotoxicity were assessed (inflammation scores 0-4; myotoxicity scores 0-6).
  • H&E hematoxylin-eosin
  • BNP@BSA-X On histological examination, the response to BNP@BSA-X was characterized by a mixed inflammatory infiltrate primarily composed of macrophages, along with a smaller population of lymphocytes and occasional neutrophils in the soft tissues surrounding the muscle #13447332v2 (inflammation; median score: 2). Furthermore, evidence of myotoxicity induced by BNP@BSA- X was observed, with degenerating and regenerating myocytes predominantly located in the perifascicular region of the muscle bundle (myotoxicity; median score: 1).
  • the strategy involves forming bupivacaine nanoparticles using the hydrophobic free base form of bupivacaine, which are then stabilized with BSA proteins.
  • the strategy involves forming diphenhydramine nanoparticles using the hydrophobic free base form of diphenhydramine, which are then stabilized with BSA proteins.
  • the strategy involves forming paclitaxel nanoparticles, which are then stabilized with BSA proteins.
  • the resulting nanoparticles are then crosslinked to create a stiff protein hydrogel.
  • the protein e.g., BSA
  • This system offers the advantage of dose flexibility for the drug. If the solvent used for drug dissolution, in this case, methanol, does not reach its saturation limit, it permits a significant increase in drug dosage. This flexibility can be particularly beneficial, especially when local anesthesia is needed for larger animals, where the minimum effective dose is higher.
  • BNP@BSA Drug binding to BSA may have had a greater effect than cross-linking per se, as evidenced by the fact that bupivacaine release was similar for BNP@BSA and BNP@BSA-X in vitro, i.e., when diffusion/dilution of BSA was prevented by containment within a dialysis membrane.
  • BNP@BSA-X resulted in much longer block durations than BNP@BSA in vivo, where disappearance of BSA from the site of injection was not limited.
  • the micron-scale pore size of BSA-X is unlikely to have a marked effect on diffusion of bupivacaine.
  • nerve block from BNP@BSA-X lasted 8 times longer than block from Exparel ® (13.3 mg/ml bupivacaine), a commercially available liposomal bupivacaine product, and had comparable or superior tissue reaction, even though it contained more bupivacaine (20 mg/ml).
  • none of the animals tested had neurobehavioral deficits in the contralateral (un-injected) hindpaw, suggesting that there was no effect from systemically distributed drug. This is important because of the potential for systemic toxicity with conventional local anesthetics.
  • BSA gel 500 mg/ml glutaraldehyde solution (GA) was added to the stirring BSA protein solution to achieve the desired final concentration of 5 mg/mL or 10 mg/mL GA.
  • BNP@BSA-X Bupivacaine free base was dissolved in methanol to prepare a stock solution.
  • Bovine serum albumin BSA was dissolved in pH 7.4 phosphate-buffered saline (PBS). The BSA solution was vigorously stirred while rapidly adding the bupivacaine free base solution.
  • DHNP@BSA-X Diphenhydramine hydrochloride was dissolved in methanol at a concentration of 100 mg/ml, and triethylamine (TEA) was added at a volume ratio of 1% (v/v). After 30 minutes of stirring, this solution was introduced into the BSA solution, previously prepared in pH 7.4 PBS at a concentration of 50 mg/ml. This addition resulted in a final diphenhydramine concentration of 1 mg/ml. Subsequently, GA was added to the solution at a final concentration of 5 mg/ml to create the diphenhydramine nanoparticle-embedded hydrogel, denoted as DHNP@BSA-X.
  • TEA triethylamine
  • BSA-X Preparation of PNP in BSA-X(BNP@BSA-X)
  • Paclitaxel was dissolved in methanol to create a stock solution with a concentration of 100 mg/ml.
  • bovine serum albumin (BSA) was dissolved in pH 7.4 phosphate- buffered saline (PBS) at a concentration of 50 mg/ml.
  • the BSA solution was vigorously stirred while the paclitaxel solution was rapidly added to reach a final paclitaxel concentration of 1 mg/ml.
  • the resulting mixture underwent rotary evaporation to remove the methanol and was continuously stirred for 2 hr to facilitate nanoparticle hardening.
  • ICG-BNP@BSA BSA
  • ICG-BNP@BSA bovine serum albumin
  • PBS pH 7.4 phosphate-buffered saline
  • the Bupivacaine free base and ICG solution were then rapidly added to the BSA solution while stirring.
  • the final concentration of BNP and ICG was 20 mg/ml and 0.05 mg/ml, respectively.
  • BNP@Cy5.5-BSA-X Cy5.5 dye conjugated BSA hydrogel
  • Bupivacaine free base was dissolved in methanol to prepare a stock solution.
  • a solution of Cy5.5 dye-conjugated BSA (Cy5.5-BSA, purchased from Nanocs Inc, New York, NY, USA) was mixed with BSA to create a mixture with a final concentration of 50 mg/ml BSA.
  • the concentration of Cy5.5 dye in the solution was 0.05 mg/ml.
  • the mixture was vigorously stirred while the bupivacaine free base was rapidly added.
  • the resulting mixture was then subjected to rotary evaporation to remove the methanol and continuously stirred for 2 hours to promote the hardening of nanoparticles.
  • the resulting solution was then treated with addition of a 500 mg/ml glutaraldehyde solution to achieve the desired final concentration of 5 mg/ml.
  • Dynamic Light Scattering The particle size was measured with a Malvern Nano ZetaSizer (Malvern Panalytical, Westborough, MA, USA). Transmission Electron Microscopy (TEM) The BNP@BSA solution was prepared by subjecting it to centrifugation to remove any excess BSA protein. The resulting pellet was then washed three times and resuspended in water.
  • Carbon-coated 400 mesh copper grids (Ted Pella, Inc., Redding, CA, USA) were subjected to #13447332v2 glow charging for 90 seconds before BNP@BSA solution were deposited on them. A volume of 5 ⁇ L of resuspended BNP@BSA was placed on the grid and allowed to settle for 5 minutes. The grid was then washed with distilled water, stained with 1% (w/w) uranyl acetate, and wicked dry with filter paper. Transmission electron microscopy (TEM) images were acquired using a Tecnai G2 Spirit BioTWIN transmission electron microscope (FEI company, Hillsboro, OR, USA) operating at an acceleration voltage of 80 kV.
  • TEM Transmission electron microscopy
  • SEM Scanning electron microscope
  • FE-SEM Field Emission Scanning Electron Microscope
  • the BSA hydrogel was prepared, flash-frozen in liquid nitrogen, and subsequently lyophilized. Prior to imaging, the sample cross-sections were obtained using a razor blade and mounted on SEM stubs with double-sided carbon tape. To enhance conductivity, the samples were further coated with an Au/Pd alloy using sputter coating techniques.
  • the rheological characteristics of the formulations were evaluated by means of a TA DHR-2 rheometer (TA instruments, NewCastle, DE, USA) furnished with a temperature regulator. A parallel plate with a diameter of 20 mm was implemented in all tests. The distance between the plates was set at 0.3 mm. The gelation time was recorded over a 5-hour period at a frequency of 6.28 rad/s and a strain of 0.1%. Frequency sweeps were conducted within the range of 0.63 to 628 rad/s with a strain of 0.1%. Step-strain oscillation tests were performed with a shear strain of 0.1% for 60 seconds, followed by 500% for 60 seconds, repeated for three cycles at a frequency of 6.28 rad/s.
  • the formulations were injected onto the sciatic nerve.
  • Neurobehavioural testing was conducted on both hindquarters. Deficits in the right (uninjected) extremity were used as an indicator of systemic drug distribution.
  • the sensory nerve blockade was assessed using a modified hotplate testing method. Briefly, the hind paws were exposed sequentially (left then right) to a 56 °C hotplate (Stoelting, Wood Dale, IL, USA), and the time the animal allowed its paw to remain on the hotplate (thermal latency) was measured. A thermal latency of 2 s indicated no nerve blockade (baseline), and a thermal latency of 12 s was considered maximal latency. Successful nerve blockade was defined as achieving a thermal latency above 7 s.
  • the hind paws were removed from the hotplate after 12 s to prevent thermal injury. Measurements were repeated three times in each animal at each time point, and the median was used for further data analysis.
  • the motor nerve block was assessed using a weight-bearing test to determine the motor strength of the rat's hindpaw. In brief, the rat was positioned with one hindpaw on a digital balance and allowed to bear its weight. The maximum weight that the rat could bear without the ankle touching the balance was recorded, and motor block was considered achieved when the motor strength was less than half-maximal. Measurements were repeated three times at each time point, and the median was used for further data analysis. The duration of sensory block was calculated as the time required for thermal latency to return to 7 s (halfway between baseline and maximal latencies).
  • the duration of motor block was defined as the time it took for weight-bearing to return to halfway between normal and maximal block.
  • Tissue harvesting and histology At 4 days after injection, rats were sacrificed to evaluate both acute and chronic inflammation and myotoxicity. The sciatic nerve was harvested along with surrounding tissues, and the dissector was blinded to the solution injected into each rat. Muscle samples were fixed in 10 % neutral buffered formalin, and standard techniques were employed to process them for histology (hematoxylin-eosin stained slides). An observer blinded to individual sample nature analyzed the slides.
  • the specimens were scored for inflammation (0-4 points) and myotoxicity (0-6 points), where inflammation severity was subjectively assessed as: 0, no inflammation; 1, peripheral inflammation; 2, deep inflammation; 3, muscular hemifascicular inflammation; 4, muscular holofascicular inflammation.
  • Myotoxicity was scored based on two characteristic features of local anesthetic myotoxicity, namely nuclear internalization and regeneration.
  • Nuclear internalization refers to myocytes with nuclei located away from their usual position at the cell periphery but with normal size and chromicity.
  • Regeneration refers to shrunken myocytes with scant eosinophilic cytoplasm and hyperchromatic nuclei.
  • Scoring ranged from 0 #13447332v2 (normal) to 6 (holofascicular regeneration), with the grade for a sample indicating the worst area (most severe damage) present on the slide.
  • the statistical analysis was performed using Prism 7 statistical software (GraphPad Software, San Diego, CA, USA), and a two-sided Student's t-test was used unless otherwise specified. Differences between groups were considered statistically significant when p ⁇ 0.05. Statistical significance was defined as follows: ns (p > 0.05), * (p ⁇ 0.05), ** (p ⁇ 0.01), *** (p ⁇ 0.001), and **** (p ⁇ 0.0001).
  • the concentration of BSA was maintained at 1 mg/ml in PBS, while the concentrations of BNP were 0.05, 0.1, and 0.2 mg/ml.
  • fluorescence was measured for 0.2 mg/ml of BNP in PBS.
  • UV-vis absorption spectra at 37°C were recorded on a UV-Vis spectrometer (Agilent Cary UV-Vis Compact Peltier, Agilent, Santa Clara, CA, USA) across a wavelength range of 190–350 nm.
  • the concentration of BSA was fixed at 1 mg/ml in PBS, and the concentrations of BNP were 0.02, 0.04, 0.06, and 0.08 mg/ml.
  • UV-Vis absorption was measured for 0.08 mg/ml of BNP in PBS. #13447332v2

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Abstract

La présente divulgation propose des compositions comprenant un hydrogel et une nanoparticule stabilisée par des protéines. L'hydrogel comprend une matrice de protéine réticulée, la nanoparticule stabilisée par des protéines comprend un agent thérapeutique ou un agent de diagnostic, et la nanoparticule stabilisée par des protéines est incorporée dans la matrice de protéine réticulée. Les compositions proposent ainsi un moyen d'administration régulée et prolongée d'agents thérapeutiques. La présente divulgation propose en outre des kits contenant les compositions, des méthodes de traitement ou de prévention d'une maladie ou d'un trouble, ainsi que des procédés de préparation des compositions.
PCT/US2024/059479 2023-12-12 2024-12-11 Hydrogels comprenant des nanoparticules stabilisées par des protéines pour une libération prolongée Pending WO2025128639A1 (fr)

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20100196478A1 (en) * 1998-09-25 2010-08-05 Gel-Del Technologies, Inc. Protein matrix materials, devices and methods of making and using thereof
US20160303281A1 (en) * 2015-04-17 2016-10-20 Rochal Industries, Llc Composition and kits for pseudoplastic microgel matrices
WO2023196970A1 (fr) * 2022-04-08 2023-10-12 University Of Pittsburgh - Of The Commonwealth System Of Higher Education Nanovésicules liées à une matrice encapsulées dans des hydrogels

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20100196478A1 (en) * 1998-09-25 2010-08-05 Gel-Del Technologies, Inc. Protein matrix materials, devices and methods of making and using thereof
US20160303281A1 (en) * 2015-04-17 2016-10-20 Rochal Industries, Llc Composition and kits for pseudoplastic microgel matrices
WO2023196970A1 (fr) * 2022-04-08 2023-10-12 University Of Pittsburgh - Of The Commonwealth System Of Higher Education Nanovésicules liées à une matrice encapsulées dans des hydrogels

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