[go: up one dir, main page]

WO2025174876A1 - Conjugaison de cystéine - Google Patents

Conjugaison de cystéine

Info

Publication number
WO2025174876A1
WO2025174876A1 PCT/US2025/015581 US2025015581W WO2025174876A1 WO 2025174876 A1 WO2025174876 A1 WO 2025174876A1 US 2025015581 W US2025015581 W US 2025015581W WO 2025174876 A1 WO2025174876 A1 WO 2025174876A1
Authority
WO
WIPO (PCT)
Prior art keywords
optionally substituted
formula
compound
peptide
salt
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
PCT/US2025/015581
Other languages
English (en)
Inventor
Kenneth SKINNER
David N. KAMBER
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Quantum Si Inc
Original Assignee
Quantum Si Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Quantum Si Inc filed Critical Quantum Si Inc
Publication of WO2025174876A1 publication Critical patent/WO2025174876A1/fr
Pending legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/06—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a carbon chain containing only aliphatic carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
    • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
    • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D213/06—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom containing only hydrogen and carbon atoms in addition to the ring nitrogen atom
    • C07D213/16—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom containing only hydrogen and carbon atoms in addition to the ring nitrogen atom containing only one pyridine ring
    • C07D213/18—Salts thereof
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
    • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
    • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D213/06—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom containing only hydrogen and carbon atoms in addition to the ring nitrogen atom
    • C07D213/16—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom containing only hydrogen and carbon atoms in addition to the ring nitrogen atom containing only one pyridine ring
    • C07D213/20—Quaternary compounds thereof
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
    • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
    • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D213/24—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with substituted hydrocarbon radicals attached to ring carbon atoms
    • C07D213/28—Radicals substituted by singly-bound oxygen or sulphur atoms
    • C07D213/32—Sulfur atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/107—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides
    • C07K1/1072—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides by covalent attachment of residues or functional groups
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/107—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides
    • C07K1/1072—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides by covalent attachment of residues or functional groups
    • C07K1/1077—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides by covalent attachment of residues or functional groups by covalent attachment of residues other than amino acids or peptide residues, e.g. sugars, polyols, fatty acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/12—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by hydrolysis, i.e. solvolysis in general
    • C07K1/128—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by hydrolysis, i.e. solvolysis in general sequencing

Definitions

  • lysine is absent in some proteins (e.g., p14ARF tumor suppressor), which cannot be sequenced by those approaches. Expanding conjugation beyond lysine residues would help streamline workflows in protein sequencing and identification.
  • a compound of Formula (I) or a salt thereof, wherein X, n, L 1 , R 1 , and Z are defined herein.
  • a compound of Formula (IV′) or a salt thereof, wherein X, n, L 1 , R B , R 1 , R 2 , R 3a , and R 3b are defined herein.
  • a compound of Formula (IV): or a salt thereof wherein X, n, L 1 , R 1 , R 2 , R 3a , and R 3b are defined herein.
  • a method of preparing a compound of Formula (III-i): or a salt thereof, comprising contacting a compound of Formula (V): or a salt thereof, with a compound of Formula (II):
  • a method of preparing a compound of Formula (VIII): or a salt thereof comprising contacting a compound of Formula (V): or a salt thereof, with a compound of Formula (IX):
  • a method of functionalizing a peptide having at least one free cysteine residue comprising reacting the peptide with a compound of Formula (II): or a salt thereof, to provide an azide-functionalized peptide, such that the sulfur atom of the cysteine residue in the azide-functionalized peptide is bonded to a group of the formula: wherein X, n, L 1 , and R 1 are defined herein.
  • a method of sequencing a peptide having at least one free cysteine residue comprising: reacting the peptide with a compound of Formula (II), Formula (VII), or Formula (IX): or a salt thereof, to provide an azide-functionalized peptide, such that the sulfur atom of the cysteine residue in the azide-functionalized peptide is bonded to a group of the formula: performing a peptide sequencing protocol; wherein X, n, L 1 , R 1 , L 2 , L 3 , and R 4 are defined herein.
  • R0708.70175WO00 4/180 #13640937v1 or a salt thereof to provide an azide-functionalized peptide, such that a sulfur atom of a cysteine residue in the azide-functionalized peptide is bonded to a group of the formula: contacting the azide-functionalized peptide with a compound comprising a complementary moiety, such that the azide moiety of the azide-functionalized peptide reacts with the complementary moiety via click chemistry to provide a triazole-linked conjugated peptide; immobilizing the triazole-linked conjugated peptide to a surface; and removing any non-immobilized peptides or portions of peptides; wherein X, n, L 1 , R 1 , L 2 , L 3 , and R 4 are defined herein.
  • a method of sequencing a peptide comprising: reacting the peptide with a vinylpyridinium compound, or a salt thereof, wherein the vinylpyridinium compound comprises an azide moiety, to provide an azide-functionalized peptide, wherein a sulfur atom of a cysteine residue in the azide-functionalized peptide is bonded to an ethylenepyridinium moiety; and performing a peptide sequencing protocol.
  • FIGs.15A-15C show GFAP conjugation of Cys with APN & VPN.
  • FIG.15A shows GFAP APN (Asp-N digest). DYRRQLQSLTC (SEQ ID NO: 6); 1882.9229 (1); 941.9651 (2); 628.3125 (3).
  • FIG.15B shows GFAP APN (Glu-C digest). DYRRQLQSLTC (SEQ ID NO: 6); 1882.9229 (1); 941.9651 (2); 628.3125 (3).
  • FIG.15C shows GFAP VPN (Asp-N digest). DYRRQLQSLTC (SEQ ID NO: 6); 1556.7751 (1); 778.8912 (2); 519.5966 (3).
  • FIGs.16A-15D show Vimentin conjugation of Cys with APN & VPN.
  • FIG.16A shows
  • FIGs.23A-23B show Vimentin alternative conjugations.
  • FIG.23A shows QL859 – GluC digest, APN.
  • FIG.23B shows representative QL859 traces, YRRQVQSLTCE (SEQ ID NO: 7) appears most abundant which is ⁇ 10 th for a typical VIME library (SOP).
  • FIGs.24A-24B show GFAP alternative conjugations.
  • FIG.24A shows QL861 – AspN digest, APN.
  • FIG.24B shows representative QL861 traces, same YRRQ peptide appears most abundant.
  • FIG.26A shows QP1112 (RLAIQFAYPFAQ) (SEQ ID NO: 8).
  • FIG.26B shows QP1149 (RLAIQFAYPACE) (SEQ ID NO: 9).
  • FIGs.27A-27B show Cys conjugation to IL1RA protein (PSGRK SSK MQAFRIWDVNQK TFYLRNNQLVAGYLQGPNVNLEEK IDVVPIEPHALFLGIHGGK MCLSCVK SGDETRLQLEAVNITDLSENRK QDK RFAFIRSDSGPTTSFESAACPGWFLCTAMEADQPVSLTNMPDEGVMVTK FYFQEDE) (SEQ ID NO: 10).
  • formulae and structures depicted herein include compounds that do not include isotopically enriched atoms, and also include compounds that include isotopically enriched atoms.
  • compounds having the present structures except for the replacement of hydrogen by deuterium or tritium, replacement of 19 F with 18 F, or the replacement of a carbon by a 13 C- or 14 C-enriched carbon are within the scope of the disclosure. Such compounds are useful, for example, as analytical tools or probes in biological assays.
  • range When a range of values (“range”) is listed, it encompasses each value and sub-range within the range. A range is inclusive of the values at the two ends of the range unless otherwise provided.
  • C 1-6 alkyl encompasses, C 1 , C 2 , C 3 , C 4 , C 5 , C 6 , C 1–6 , C 1–5 , C 1–4 , C 1–3 , C 1–2 , C 2–6 , C 2–5 , C 2–4 , C 2–3 , C 3–6 , C 3–5 , C 3–4 , C 4–6 , C 4–5 , and C 5–6 alkyl.
  • aliphatic refers to alkyl, alkenyl, alkynyl, and carbocyclic groups.
  • heteroaliphatic refers to heteroalkyl, heteroalkenyl, heteroalkynyl, and heterocyclic groups.
  • alkyl refers to a radical of a straight-chain or branched saturated hydrocarbon group having from 1 to 20 carbon atoms (“C 1–20 alkyl”). In some embodiments, an alkyl group has 1 to 12 carbon atoms (“C 1–12 alkyl”). In some embodiments, 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”).
  • an alkyl group has 1 to 8 carbon atoms (“C 1–8 alkyl”). In some embodiments, an alkyl group has 1 to 7 carbon atoms (“C 1–7 alkyl”). In some embodiments, an alkyl group has 1 to 6 carbon
  • 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”). In some embodiments, an alkyl group has 1 to 3 carbon atoms (“C1–3 alkyl”). In some embodiments, an alkyl group has 1 to 2 carbon atoms (“C 1–2 alkyl”). In some embodiments, an alkyl group has 1 carbon atom (“C 1 alkyl”). In some embodiments, an alkyl group has 2 to 6 carbon atoms (“C2-6 alkyl”).
  • C1–6 alkyl groups include methyl (C1), ethyl (C2), propyl (C3) (e.g., n-propyl, isopropyl), butyl (C4) (e.g., n-butyl, tert-butyl, sec-butyl, isobutyl), pentyl (C5) (e.g., n-pentyl, 3-pentanyl, amyl, neopentyl, 3-methyl-2-butanyl, tert- amyl), and hexyl (C6) (e.g., n-hexyl).
  • C1–6 alkyl groups include methyl (C1), ethyl (C2), propyl (C3) (e.g., n-propyl, isopropyl), butyl (C4) (e.g., n-butyl, tert-butyl, sec-butyl, isobuty
  • alkyl groups include n-heptyl (C7), n-octyl (C8), n-dodecyl (C12), and the like. Unless otherwise specified, each instance of an alkyl group is independently unsubstituted (an “unsubstituted alkyl”) or substituted (a “substituted alkyl”) with one or more substituents (e.g., halogen, such as F).
  • substituents e.g., halogen, such as F
  • the alkyl group is an unsubstituted C1–12 alkyl (such as unsubstituted C1–6 alkyl, e.g., ⁇ CH3 (Me), unsubstituted ethyl (Et), unsubstituted propyl (Pr, e.g., unsubstituted n-propyl (n-Pr), unsubstituted isopropyl (i-Pr)), unsubstituted butyl (Bu, e.g., unsubstituted n-butyl (n-Bu), unsubstituted tert-butyl (tert-Bu or t-Bu), unsubstituted sec-butyl (sec- Bu or s-Bu), unsubstituted isobutyl (i-Bu)).
  • unsubstituted C1–12 alkyl such as unsubstituted C1–6 alkyl, e.g.
  • the alkyl group is a substituted C1–12 alkyl (such as substituted C1–6 alkyl, e.g., –CH2F, –CHF2, –CF3, –CH2CH2F, –CH2CHF2, –CH2CF3, or benzyl (Bn)).
  • heteroalkyl refers to an alkyl group, which further includes at least one heteroatom (e.g., 1, 2, 3, or 4 heteroatoms) selected from oxygen, nitrogen, or sulfur within (e.g., inserted between adjacent carbon atoms of) and/or placed at one or more terminal position(s) of the parent chain.
  • a heteroalkyl group refers to a saturated group having from 1 to 20 carbon atoms and 1 or more heteroatoms within the parent chain (“heteroC1–20 alkyl”). In certain embodiments, a heteroalkyl group refers to a saturated group having from 1 to 12 carbon atoms and 1 or more heteroatoms within the parent chain (“heteroC1–12 alkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 11 carbon atoms and 1 or more heteroatoms within the parent chain (“heteroC 1–11 alkyl”).
  • a heteroalkyl group is a saturated group having 1 to 10 carbon atoms and 1 or more heteroatoms within the parent chain (“heteroC 1–10 alkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 9 carbon atoms and 1 or more heteroatoms within the parent chain (“heteroC 1–9 alkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 8 carbon atoms and 1 or more heteroatoms within the parent chain (“heteroC 1–8 alkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 7 carbon atoms and 1 or more heteroatoms within the parent chain (“heteroC 1–7 alkyl”).
  • an alkenyl group has 1 to 11 carbon atoms (“C1–11 alkenyl”). In some embodiments, an alkenyl group has 1 to 10 carbon atoms (“C1–10 alkenyl”). In some embodiments, an alkenyl group has 1 to 9 carbon atoms (“C1–9 alkenyl”). In some embodiments, an alkenyl group has 1 to 8 carbon atoms (“C1–8 alkenyl”). In some embodiments, an alkenyl group has 1 to 7 carbon atoms (“C1–7 alkenyl”). In some embodiments, an alkenyl group has 1 to 6 carbon atoms (“C1–6 alkenyl”).
  • C1-4 alkynyl groups include, without limitation, methylidynyl (C1), ethynyl (C2), 1-propynyl (C3), 2-propynyl (C3), 1-butynyl (C4), 2-butynyl (C4), and the like.
  • C1-6 alkenyl groups include the aforementioned C2-4 alkynyl groups as well as pentynyl (C5), hexynyl (C6), and the like.
  • Additional examples of alkynyl include heptynyl (C7), octynyl (C8), and the like.
  • each 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 C1-20 alkynyl.
  • the alkynyl group is a substituted C1-20 alkynyl.
  • Exemplary C 3-8 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 (C 8 ), bicyclo[2.2.1]heptanyl (C 7 ), bicyclo[2.2.2]octanyl (C 8 ), and the like.
  • Exemplary C 3-8 carbocyclyl groups include the aforementioned C 3-10 carbocyclyl groups as well as cycloundecyl (C 11 ), spiro[5.5]undecanyl (C 11 ), cyclododecyl (C 12 ), cyclododecenyl (C 12 ), cyclotridecane (C 13 ), cyclotetradecane (C 14 ), and the like.
  • a heterocyclyl group can either be monocyclic (“monocyclic heterocyclyl”) or polycyclic (e.g., a fused, bridged or spiro ring system such as a bicyclic system (“bicyclic heterocyclyl”) or tricyclic system (“tricyclic heterocyclyl”)), and can be saturated or can contain one or more carbon-carbon double or triple bonds.
  • Heterocyclyl polycyclic ring systems can include one or more heteroatoms in one or both rings.
  • Heterocyclyl also includes ring systems wherein the heterocyclyl ring, as defined above, is fused with one or more carbocyclyl groups wherein the point of attachment is either on the carbocyclyl or heterocyclyl ring, or ring systems wherein the heterocyclyl ring, as defined above, is fused with one or more aryl or heteroaryl groups, wherein the point of attachment is on the heterocyclyl ring, and in such instances, the number of ring members continue to designate the number of ring members in the heterocyclyl ring system. Unless otherwise specified, each instance of
  • 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”).
  • a heterocyclyl group is a 5–6 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 (“5–6 membered heterocyclyl”).
  • the 5–6 membered heterocyclyl has 1–3 ring heteroatoms selected from nitrogen, oxygen, and sulfur.
  • Exemplary 5-membered heterocyclyl groups containing 1 heteroatom include tetrahydrofuranyl, dihydrofuranyl, tetrahydrothiophenyl, dihydrothiophenyl, pyrrolidinyl, dihydropyrrolyl, and pyrrolyl-2,5-dione.
  • Exemplary 5-membered heterocyclyl groups containing 2 heteroatoms include dioxolanyl, oxathiolanyl and dithiolanyl.
  • Exemplary 5-membered heterocyclyl groups containing 3 heteroatoms include triazolinyl, oxadiazolinyl, and thiadiazolinyl.
  • each instance of an aryl group is independently unsubstituted (an “unsubstituted aryl”) or substituted (a “substituted aryl”) with one or more substituents.
  • the aryl group is an unsubstituted C6-14 aryl.
  • the aryl group is a substituted C6-14 aryl.
  • heteroaryl refers to a radical of a 5-14 membered monocyclic or polycyclic (e.g., bicyclic, tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10, or 14 ⁇ electrons shared in a cyclic array) having ring carbon atoms and 1–4 ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-14 membered heteroaryl”).
  • 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.
  • Heteroaryl includes ring systems wherein the heteroaryl ring, as defined above, is fused with one or more carbocyclyl or heterocyclyl groups wherein the point of attachment is on the heteroaryl ring, and in such instances, the number of ring members continue to designate the number of ring members in the heteroaryl ring system. “Heteroaryl” also includes ring systems wherein the heteroaryl ring, as defined above, is fused with one or more aryl groups wherein the point of attachment is either on the aryl or heteroaryl ring, and in such instances, the number of ring members designates the number of ring members in the fused polycyclic (aryl/heteroaryl) ring system.
  • 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
  • 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. In some embodiments, 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.
  • 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 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 6-membered heteroaryl groups containing 2 heteroatoms include pyridazinyl, pyrimidinyl, and pyrazinyl.
  • Exemplary 6-membered heteroaryl groups containing 3 or 4 heteroatoms include triazinyl and tetrazinyl, respectively.
  • Exemplary 7-membered heteroaryl groups containing 1 heteroatom include azepinyl, oxepinyl, and thiepinyl.
  • 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.
  • 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. [0064] 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.
  • alkylene is the divalent moiety of alkyl
  • alkenylene is the divalent moiety of alkenyl
  • alkynylene is the divalent moiety of alkynyl
  • heteroalkylene is the divalent moiety of heteroalkyl
  • heteroalkenylene is the divalent moiety of heteroalkenyl
  • heteroalkynylene is the divalent moiety of heteroalkynyl
  • carbocyclylene is the divalent moiety
  • heterocyclylene is the divalent moiety of heterocyclyl
  • arylene is the divalent moiety of aryl
  • heteroarylene is the divalent moiety of heteroaryl.
  • a group is optionally substituted unless expressly provided otherwise.
  • the term “optionally substituted” refers to being substituted or unsubstituted.
  • alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl groups are optionally substituted.
  • Optionally substituted refers to a group which is substituted or unsubstituted (e.g., “substituted” or “unsubstituted” alkyl, “substituted” or “unsubstituted” alkenyl, “substituted” or “unsubstituted” alkynyl, “substituted” or “unsubstituted” heteroalkyl, “substituted” or “unsubstituted” heteroalkenyl, “substituted” or “unsubstituted” heteroalkynyl, “substituted” or “unsubstituted” carbocyclyl, “substituted” or “unsubstituted” heterocyclyl, “substituted” or “unsubstituted” aryl or “substituted” or “unsubstituted” heteroaryl group).
  • substituted means that at least one hydrogen present on a group is replaced with a permissible substituent, e.g., a substituent which upon substitution results in a stable compound, e.g., a compound which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, or other reaction.
  • a “substituted” group has a substituent at one or more substitutable positions of the group, and when more than one position in any given structure is substituted, the substituent is either the same or different at each position.
  • substituted is contemplated to include substitution with all permissible substituents of organic compounds, and includes any of the substituents described herein that results in the formation of a stable compound.
  • the present disclosure contemplates any and all such combinations in order to arrive at a stable compound.
  • heteroatoms such as nitrogen may have hydrogen substituents and/or any suitable substituent as described herein which satisfy the valencies of the heteroatoms and results in the formation of a stable moiety.
  • the disclosure is not limited in any manner by the exemplary substituents described herein.
  • R0708.70175WO00 17/180 #13640937v1 X ⁇ is a counterion; each instance of R ee is, independently, selected from C1–10 alkyl, C1–10 perhaloalkyl, C1–10 alkenyl, C1–10 alkynyl, heteroC1–10 alkyl, heteroC1–10 alkenyl, heteroC1–10 alkynyl, C3-10 carbocyclyl, C 6-10 aryl, 3-10 membered heterocyclyl, and 3-10 membered heteroaryl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 R gg groups; each instance of R ff is, independently, selected from hydrogen, C1–10 alkyl, C1–10 perhaloalkyl, C1
  • each carbon atom substituent is independently halogen, substituted (e.g., substituted with one or more
  • 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 C1–10 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-
  • a carbon atom substituent consists of carbon, hydrogen, fluorine, and/or chlorine atoms.
  • halo or “halogen” refers to fluorine (fluoro, ⁇ F), chlorine (chloro, ⁇ Cl), bromine (bromo, ⁇ Br), or iodine (iodo, ⁇ I).
  • hydroxyl or “hydroxy” refers to the group ⁇ OH.
  • Nitrogen atoms can be substituted or unsubstituted as valency permits, and include primary, secondary, tertiary, and quaternary nitrogen atoms.
  • each nitrogen protecting group 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-trichloroethyl carbamate (Troc), 2-trimethylsilylethyl carbamate (Teoc), 2- phenylethyl carbamate (hZ), 1–(1-adamantyl)-1-methylethyl carba
  • 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
  • each nitrogen protecting group is independently selected from the group 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-di
  • 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.
  • Oxygen protecting groups are well known in the art and include those described in detail in Protecting Groups in Organic Synthesis, T. W. Greene and P. G. M. Wuts, 3 rd edition, John Wiley & Sons, 1999, incorporated herein by reference.
  • 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-bromotetrahydropyranyl, tetrahydrothiopyranyl, 1-methoxycyclo
  • 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
  • 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. [0093] A “counterion” or “anionic counterion” is a negatively charged group associated with a positively charged group in order to maintain electronic neutrality.
  • the compound of Formula (I′) is of Formula (I). [0117] In some embodiments, the compound of Formula (I) is of Formula (II): or a salt thereof. [0118] In some embodiments, the compound of Formula (II′) is of Formula (II).
  • the compound of Formula (I) is of Formula (III): or a salt thereof.
  • the compound of Formula (III′) is of Formula (III).
  • R 2 is optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; and each of R 3a and R 3b is independently hydrogen, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl, or R 3a and R 3b are joined together with their intervening atoms to form optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl.
  • L 1 comprises substituted linear alkylene. In some embodiments, L 1 comprises substituted linear C1-C12 alkylene. In some embodiments, L 1 comprises substituted linear C1-C6 alkylene. In some embodiments, L 1 comprises substituted linear C1-C3 alkylene. [0133] In some embodiments, L 1 comprises unsubstituted alkylene. In some embodiments, L 1 comprises unsubstituted C1-C12 alkylene. In some embodiments, L 1 comprises unsubstituted C1-C6 alkylene. In some embodiments, L 1 comprises unsubstituted C1-C3 alkylene. In some embodiments, L 1 comprises unsubstituted linear alkylene.
  • L 1 comprises substituted C1-C6 alkenylene. In some embodiments, L 1 comprises substituted C1-C3 alkenylene. In some embodiments, L 1 comprises unsubstituted alkenylene. In some embodiments, L 1 comprises unsubstituted C1-C12 alkenylene. In some embodiments, L 1 comprises unsubstituted C1-C6 alkenylene. In some embodiments, L 1 comprises unsubstituted C1-C3 alkenylene. [0137] In some embodiments, L 1 comprises optionally substituted alkynylene. In some embodiments, L 1 comprises optionally substituted C1-C12 alkynylene.
  • L 1 comprises unsubstituted C1-C6 alkynylene. In some embodiments, L 1 comprises unsubstituted C1-C3 alkynylene. [0138] In some embodiments, L 1 comprises optionally substituted heteroalkylene. In some embodiments, L 1 comprises optionally substituted C1-C12 heteroalkylene. In some embodiments, L 1 comprises optionally substituted C1-C6 heteroalkylene. In some embodiments, L 1 comprises optionally substituted C1-C3 heteroalkylene. In some embodiments, L 1 comprises optionally substituted linear heteroalkylene. In some embodiments, L 1 comprises optionally substituted linear C 1 -C 12 heteroalkylene.
  • L 1 comprises optionally substituted linear C 1 -C 6 heteroalkylene. In some embodiments, L 1 comprises optionally substituted linear C 1 -C 3 heteroalkylene. [0139] In some embodiments, L 1 comprises substituted heteroalkylene. In some embodiments, L 1 comprises substituted C 1 -C 12 heteroalkylene. In some embodiments, L 1 comprises substituted C 1 -C 6 heteroalkylene. In some embodiments, L 1 comprises substituted C 1 -C 3 heteroalkylene. In some embodiments, L 1 comprises substituted linear heteroalkylene. In some embodiments, L 1 comprises substituted linear C 1 -C 12 heteroalkylene. In some embodiments, L 1 comprises substituted linear C 1 -C 6 heteroalkylene.
  • L 1 comprises unsubstituted linear C 1 -C 3 heteroalkylene.
  • L 1 comprises , wherein x is an integer between 1 and 100, inclusive.
  • L 1 is or , wherein x is an integer between 1 and 100, inclusive.
  • x is an integer between 1 and 100, inclusive.
  • x is an integer between 1 and 90, inclusive; between 1 and 80, inclusive; between 1 and 70, inclusive; between 1 and 60, inclusive; between 1 and 50, inclusive; between 1 and 40, inclusive; between 1 and 30, inclusive; between 1 and 20, inclusive; between 1 and 10, inclusive; or between 1 and 5, inclusive.
  • x is an integer between 1 and 10, inclusive.
  • x is an integer between 1 and 5, inclusive.
  • x is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.
  • x is 1, 2, 3, 4, 5, 6, 7, 8, or 9.
  • x is 1, 2, 3, 4, 5, 6, 7, or 8.
  • x is 1, 2, 3, 4, 5, 6, or 7.
  • x is 1, 2, 3, 4, 5, or 6. In some embodiments, x is 1, 2, 3, 4, or 5. In some embodiments, x is 1, 2, 3, or 4. In some embodiments, x is 1, 2, or 3. In some embodiments, x is 1 or 2. In some embodiments, x is 1. In some embodiments, x is 2. In some embodiments, x is 3. In some embodiments, x is 4. In some embodiments, x is 5. In some embodiments, x is 6. In some embodiments, x is 7. In some embodiments, x is 8. In some embodiments, x is 9. In some embodiments, x is 10. [0143] In some embodiments, L 1 comprises or , wherein x is an integer between 1 and 10, inclusive.
  • L 1 comprises or embodiments, L 1 is , wherein x is an integer between 1 and 5, inclusive.
  • L 1 comprises optionally substituted heteroalkenylene.
  • L 1 comprises optionally substituted C1-C12 heteroalkenylene.
  • L 1 comprises optionally substituted C1-C6 heteroalkenylene.
  • L 1 comprises optionally substituted C1-C3 heteroalkenylene.
  • L 1 comprises substituted heteroalkenylene.
  • L 1 comprises optionally substituted heteroalkynylene. In some embodiments, L 1 comprises optionally substituted C1-C12 heteroalkynylene. In some embodiments, L 1 comprises optionally substituted C1-C6 heteroalkynylene. In some embodiments, L 1 comprises optionally substituted C1-C3 heteroalkynylene. In some embodiments, L 1 comprises substituted heteroalkynylene. In some embodiments, L 1 comprises substituted C1-C12 heteroalkynylene. In some embodiments, L 1 comprises substituted C1-C6 heteroalkynylene. In some embodiments, L 1 comprises substituted C1-C3 heteroalkynylene.
  • At least one occurrence of R 1 is optionally substituted C1-C12 alkynyl. In some embodiments, at least one occurrence of R 1 is optionally substituted C1-C6 alkynyl. In some embodiments, at least one occurrence of R 1 is optionally substituted C1-C3 alkynyl. [0153] In some embodiments, at least one occurrence of R 1 is optionally substituted heteroaliphatic. In some embodiments, at least one occurrence of R 1 is optionally substituted heteroalkyl. In some embodiments, at least one occurrence of R 1 is optionally substituted C1-C12 heteroalkyl.
  • At least one occurrence of R 1 is optionally substituted C 1 -C 12 heteroalkynyl. In some embodiments, at least one occurrence of R 1 is optionally substituted C 1 -C 6 heteroalkynyl. In some embodiments, at least one occurrence of R 1 is optionally substituted C 1 -C 3 heteroalkynyl. [0154] In some embodiments, at least one occurrence of R 1 is optionally substituted carbocyclyl. In some embodiments, at least one occurrence of R 1 is optionally substituted C 3 -C 10 carbocyclyl. In some embodiments, at least one occurrence of R 1 is optionally substituted C 3 -C 6 carbocyclyl.
  • each occurrence of R A is independently hydrogen, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, a nitrogen protecting group when attached to a nitrogen atom, an oxygen protecting group when attached to an oxygen atom, or a sulfur protecting group when attached to a sulfur atom, or two occurrences of R A are joined together with their intervening atom(s) to form an optionally substituted heterocyclic ring or optionally substituted heteroaryl ring; [0158] In some embodiments, at least one occurrence of R A is hydrogen, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, a nitrogen protecting group when attached to a nitrogen atom, an oxygen protecting group when attached to an oxygen atom
  • R 2 is optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl.
  • R 2 is optionally substituted aliphatic. In some embodiments, R 2 is
  • R0708.70175WO00 48/180 #13640937v1 optionally substituted alkyl.
  • R 2 is optionally substituted C 1 -C 12 alkyl.
  • R 2 is optionally substituted C1-C6 alkyl.
  • R 2 is optionally substituted C1-C3 alkyl.
  • R 2 is optionally substituted linear alkyl. [0172] In some embodiments, R 2 is substituted alkyl. In some embodiments, R 2 is substituted C 1 -C 12 alkyl. In some embodiments, R 2 is substituted C1-C6 alkyl. In some embodiments, R 2 is substituted C1-C3 alkyl.
  • R 2 is substituted linear alkyl.
  • R 2 is optionally substituted alkenyl. In some embodiments, R 2 is optionally substituted C1-C12 alkenyl. In some embodiments, R 2 is optionally substituted C1-C6 alkenyl. In some embodiments, R 2 is optionally substituted C1-C3 alkenyl. In some embodiments, R 2 is optionally substituted alkynyl. In some embodiments, R 2 is optionally substituted C1-C12 alkynyl. In some embodiments, R 2 is optionally substituted C 1 -C 6 alkynyl. In some embodiments, R 2 is optionally substituted C 1 -C 3 alkynyl.
  • R 2 is optionally substituted heteroaliphatic. In some embodiments, R 2 is optionally substituted heteroalkyl. In some embodiments, R 2 is optionally substituted C 1 -C 12 heteroalkyl. In some embodiments, R 2 is optionally substituted C 1 -C 6 heteroalkyl. In some embodiments, R 2 is optionally substituted C 1 -C 3 heteroalkyl. [0177] In some embodiments, R 2 is substituted heteroaliphatic. In some embodiments, R 2 is substituted heteroalkyl. In some embodiments, R 2 is substituted C 1 -C 12 heteroalkyl. In some embodiments, R 2 is substituted C 1 -C 6 heteroalkyl.
  • R 2 is substituted C 1 -C 3 heteroalkyl.
  • R N is hydrogen, a nitrogen protecting group, an amino acid moiety, or a peptide. In some embodiments, R N is hydrogen. In some embodiments, R N is a nitrogen protecting group. In some embodiments, R N is an amino acid moiety or a peptide. In some embodiments, R N is an amino acid moiety. In some embodiments, R N is a peptide. [0184] In some embodiments, R C is -OH, and R N is an amino acid moiety or a peptide. In some embodiments, R C is -OH, and R N is an amino acid moiety. In some embodiments, R C is -OH, and R N is a peptide.
  • R C is an amino acid moiety or a peptide, and R N is hydrogen. In some embodiments, R C is an amino acid moiety, and R N is hydrogen. In some embodiments, R C is a peptide, and R N is hydrogen. [0185] In some embodiments, R C is an amino acid moiety or a peptide; and R N is an amino acid moiety or a peptide. In some embodiments, R C is an amino acid moiety; and R N is an amino acid moiety. In some embodiments,
  • R 2 is optionally substituted carbocyclyl. In some embodiments, R 2 is optionally substituted C3-C10 carbocyclyl. In some embodiments, R 2 is optionally substituted C3-C6 carbocyclyl. In some embodiments, R 2 is optionally substituted heterocyclyl. In some embodiments, R 2 is optionally substituted 3-10 membered heterocyclyl. In some embodiments, R 2 is optionally substituted 3- 6 membered heterocyclyl. In some embodiments, R 2 is optionally substituted aryl. In some embodiments, R 2 is optionally substituted phenyl. In some embodiments, R 2 is optionally substituted heteroaryl. In some
  • At least one of R 3a and R 3b is substituted C 1 -C 12 alkyl. In some embodiments, at least one of R 3a and R 3b is substituted C 1 -C 6 alkyl. In some embodiments, at least one of R 3a and R 3b is substituted C 1 -C 3 alkyl. In some embodiments, at least one of R 3a and R 3b is substituted linear alkyl. In some embodiments, R 3a is optionally substituted alkyl. In some embodiments, R 3a is optionally substituted C 1 -C 12 alkyl. In some embodiments, R 3a is optionally substituted C 1 -C 6 alkyl.
  • R 3a is optionally substituted C1-C3 alkyl. In some embodiments, R 3a is optionally substituted linear alkyl. In some embodiments, R 3a is substituted alkyl. In some embodiments, R 3a is substituted C1-C12 alkyl. In some embodiments, R 3a is substituted C1-C6 alkyl. In some embodiments, R 3a is substituted C1-C3 alkyl. In some embodiments, R 3a is substituted linear alkyl. In some embodiments, R 3b is optionally substituted alkyl. In some embodiments, R 3b is optionally substituted C1-C12 alkyl. In some embodiments, R 3b is optionally substituted C1-C6 alkyl.
  • At least one of R 3a and R 3b is optionally substituted C1-C6 alkenyl. In some embodiments, at least one of R 3a and R 3b is optionally substituted C1-C3 alkenyl. In some embodiments, at least one of R 3a and R 3b is
  • R 3b is substituted alkenyl. In some embodiments, R 3b is substituted C1-C12 alkenyl. In some embodiments, R 3b is substituted C1-C6 alkenyl. In some embodiments, R 3b is substituted C1-C3 alkenyl. [0197] In some embodiments, at least one of R 3a and R 3b is optionally substituted alkynyl. In some embodiments, at least one of R 3a and R 3b is optionally substituted C1-C12 alkynyl. In some embodiments, at least one of R 3a and R 3b is optionally substituted C1-C6 alkynyl.
  • R 3a is substituted heteroalkenyl. In some embodiments, R 3a is substituted C 1 -C 12 heteroalkenyl. In some embodiments, R 3a is substituted C 1 -C 6 heteroalkenyl. In some embodiments, R 3a is substituted C 1 -C 3 heteroalkenyl. In some embodiments, R 3b is optionally substituted heteroalkenyl. In some embodiments, R 3b is optionally substituted C 1 -C 12 heteroalkenyl. In some embodiments, R 3b is optionally substituted C 1 -C 6 heteroalkenyl. In some embodiments, R 3b is optionally substituted C 1 -C 3 heteroalkenyl.
  • At least one of R 3a and R 3b is optionally substituted carbocyclyl. In some embodiments, at least one of R 3a and R 3b is optionally substituted C3-C10 carbocyclyl. In some embodiments, at least one of R 3a and R 3b is optionally substituted C3-C6 carbocyclyl. In some embodiments, at least one of R 3a and R 3b is substituted carbocyclyl. In some embodiments, at least one of R 3a and R 3b is substituted C3-C10 carbocyclyl. In some embodiments, at least one of R 3a and R 3b is substituted C3-C6 carbocyclyl.
  • x is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.
  • the compound of Formula (III) is of Formula (III-C-i-a) or Formula (III- D-i-a): or a salt thereof, wherein x is an integer between 1 and 100, inclusive.
  • x is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.
  • R 3a and R 3b are joined together with their intervening atoms to form optionally substituted carbocyclyl or optionally substituted heterocyclyl. In some embodiments of Formula (IV-C) or Formula (IV-D), R 3a and R 3b are joined together with their intervening atoms to form substituted carbocyclyl or substituted heterocyclyl. [0228] In some embodiments, the compound of Formula (IV) is of Formula (IV-C-i) or Formula (IV-D- i):
  • the molar ratio of the compound of Formula (II), or salt thereof, to the compound of Formula (V), or salt thereof is between about 1:1 and about 50:1, about 1:1 and about 45:1, about 1:1 and about 40:1, about 1:1 and about 35:1, about 1:1 and about 30:1, about 1:1 and about 25:1, about 1:1 and about 20:1, about 1:1 and about 15:1, about 1:1 and about 10:1, about 1:1 and about 5:1, about 5:1 and about 30:1, about 5:1 and about 25:1, about 5:1 and about 20:1, about 5:1 and about 15:1, about 5:1 and about 10:1, about 10:1 and about 30:1, about 10:1 and about 25:1, about 10:1 and about 20:1, about 10:1 and about 15:1, about 15:1 and about 30:1, about 15:1 and about 25:1, or about 15:1 and about 20:1. In some embodiments, the molar ratio of the compound of Formula (II), or salt thereof, to the compound of Formula (V), or salt thereof, is between about 1:1 and about 50:1,
  • a molar ratio of the compound of Formula (II), or salt thereof, to the compound of Formula (V), or salt thereof is between about 1:1 and about 50:1. In some embodiments, the molar ratio of the compound of Formula (II), or salt thereof, to the compound of Formula (V), or salt thereof, is between about 5:1 and about 25:1.
  • L 1 comprises substituted linear C 1 -C 12 alkylene. In some embodiments, L 1 comprises substituted linear C 1 -C 6 alkylene. In some embodiments, L 1 comprises substituted linear C 1 -C 3 alkylene. [0238] In some embodiments, L 1 comprises unsubstituted alkylene. In some embodiments, L 1 comprises
  • L 1 comprises unsubstituted C 1 -C 6 alkylene. In some embodiments, L 1 comprises unsubstituted C1-C3 alkylene. In some embodiments, L 1 comprises unsubstituted linear alkylene. In some embodiments, L 1 comprises unsubstituted linear C1-C12 alkylene. In some embodiments, L 1 comprises unsubstituted linear C 1 -C 6 alkylene. In some embodiments, L 1 comprises unsubstituted linear C1-C3 alkylene.
  • L 1 comprises methylene, ethylene, n-propylene, n-butylene, n-pentylene, or n-hexylene. In some embodiments, L 1 comprises methylene, ethylene, or n-propylene. In some embodiments, L 1 comprises ethylene. [0240] In some embodiments, L 1 is methylene, ethylene, n-propylene, n-butylene, n-pentylene, or n- hexylene. In some embodiments, L 1 is methylene, ethylene, or n-propylene. In some embodiments, L 1 is ethylene. [0241] In some embodiments, L 1 comprises optionally substituted alkenylene.
  • L 1 comprises optionally substituted C1-C12 alkenylene. In some embodiments, L 1 comprises optionally substituted C1-C6 alkenylene. In some embodiments, L 1 comprises optionally substituted C1-C3 alkenylene. In some embodiments, L 1 comprises substituted alkenylene. In some embodiments, L 1 comprises substituted C1-C12 alkenylene. In some embodiments, L 1 comprises substituted C1-C6 alkenylene. In some embodiments, L 1 comprises substituted C1-C3 alkenylene. In some embodiments, L 1 comprises unsubstituted alkenylene. In some embodiments, L 1 comprises unsubstituted C1-C12 alkenylene.
  • L 1 comprises unsubstituted C1-C6 heteroalkylene. In some embodiments, L 1 comprises unsubstituted C1-C3 heteroalkylene. In some embodiments, L 1 comprises unsubstituted linear heteroalkylene. In some embodiments, L 1 comprises unsubstituted linear C1-C12 heteroalkylene. In some embodiments, L 1 comprises unsubstituted linear C1- C6 heteroalkylene. In some embodiments, L 1 comprises unsubstituted linear C1-C3 heteroalkylene. [0246] In some embodiments, L 1 comprises , wherein x is an integer between 1 and 100, inclusive.
  • x is 1, 2, 3, 4, 5, 6, or 7. In some embodiments, x is 1, 2, 3, 4, 5, or 6. In some embodiments, x is 1, 2, 3, 4, or 5. In some embodiments, x is 1, 2, 3, or 4. In some embodiments, x is 1, 2, or 3. In some embodiments, x is 1 or 2. In some embodiments, x is 1. In some embodiments, x is 2. In some embodiments, x is 3. In some embodiments, x is 4. In some embodiments, x is 5. In some embodiments, x is 6. In some embodiments, x is 7. In some embodiments, x is 8. In some embodiments, x is 9. In some embodiments, x is 10. [0248] In some embodiments, L 1 comprises , wherein x is an integer between 1 and 10, inclusive. In some embodiments, L 1 comprises or
  • R C is an amino acid moiety; and R N is an amino acid moiety. In some embodiments, R C is an amino acid moiety; and R N is a peptide. In some embodiments, R C is a peptide; and R N is an amino acid moiety. In some embodiments, R C is a peptide; and R N is a peptide.
  • the compound of Formula (II) is of Formula (II-A) or Formula (II-B): or a salt thereof, wherein x is an integer between 1 and 100, inclusive. In some embodiments of Formula (II-B), x is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.
  • R C is an amino acid moiety or a peptide.
  • R N is an amino acid moiety or a peptide.
  • R C is an amino acid moiety or a peptide; and R N is an amino acid moiety or a peptide.
  • R C is -OH, and R N is an amino acid moiety. In some embodiments, R C is -OH, and R N is a peptide. In some embodiments, R C is an amino acid moiety or a peptide, and R N is hydrogen. In some embodiments, R C is an amino acid moiety, and R N is hydrogen. In some embodiments, R C is a peptide, and R N is hydrogen. In some embodiments, R C is an amino acid moiety or a peptide; and R N is an amino acid moiety or a peptide. In some embodiments, R C is an amino acid moiety; and R N is an amino acid moiety. In some embodiments, R C is an amino acid moiety; and R N is an amino acid moiety.
  • L 3 is optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted heteroalkylene, optionally substituted heteroalkenylene, optionally substituted heteroalkynylene, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, optionally substituted heteroarylene, or a combination thereof.
  • L 3 comprises optionally substituted alkylene.
  • L 3 comprises optionally substituted C1-C12 alkylene.
  • L 3 comprises optionally substituted C1-C6 alkylene. In some embodiments, L 3 comprises optionally substituted C1-C3 alkylene. In some embodiments, L 3 comprises optionally substituted linear alkylene. In some embodiments, L 3 comprises optionally substituted linear C1-C12 alkylene. In some embodiments, L 3 comprises optionally substituted linear C1-C6 alkylene. In some embodiments, L 3 comprises optionally substituted linear C1-C3 alkylene. [0300] In some embodiments, L 3 comprises substituted alkylene. In some embodiments, L 3 comprises substituted C1-C12 alkylene. In some embodiments, L 3 comprises substituted C1-C6 alkylene.
  • L 3 comprises unsubstituted C1-C6 alkynylene. In some embodiments, L 3 comprises unsubstituted C1-C3 alkynylene. [0304] In some embodiments, L 3 comprises optionally substituted heteroalkylene. In some embodiments, L 3 comprises optionally substituted C1-C12 heteroalkylene. In some embodiments, L 3 comprises optionally substituted C1-C6 heteroalkylene. In some embodiments, L 3 comprises optionally substituted C1-C3 heteroalkylene. In some embodiments, L 3 comprises optionally substituted linear heteroalkylene. In some embodiments, L 3 comprises optionally substituted linear C1-C12 heteroalkylene.
  • L 3 comprises unsubstituted linear C 1 -C 3 heteroalkylene. [0307] In some embodiments, L 3 comprises , wherein z is an integer between 1 and 100, inclusive. [0308] As generally described herein, z is an integer between 1 and 100, inclusive. In some embodiments, z is an integer between 1 and 90, inclusive; between 1 and 80, inclusive; between 1 and 70, inclusive; between 1 and 60, inclusive; between 1 and 50, inclusive; between 1 and 40, inclusive; between 1 and 30, inclusive; between 1 and 20, inclusive; between 1 and 10, inclusive; or between 1 and 5, inclusive. In some embodiments, z is an integer between 1 and 10, inclusive. In some embodiments, z is an integer between 1 and 5, inclusive.
  • z is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, z is 1, 2, 3, 4, 5, 6, 7, 8, or 9. In some embodiments, z is 1, 2, 3, 4, 5, 6, 7, or 8. In some embodiments, z is 1, 2, 3, 4, 5, 6, or 7. In some embodiments, z is 1, 2, 3, 4, 5, or 6. In some embodiments, z is 1, 2, 3, 4, or 5. In some embodiments, z is 1, 2, 3, or 4. In some embodiments, z is 1, 2,
  • z is 1 or 2. In some embodiments, z is 1. In some embodiments, z is 2. In some embodiments, z is 3. In some embodiments, z is 4. In some embodiments, z is 5. In some embodiments, z is 6. In some embodiments, z is 7. In some embodiments, z is 8. In some embodiments, z is 9. In some embodiments, z is 10. [0309] In some embodiments, L 3 comprises , wherein z is an integer between 1 and 10, inclusive. In some embodiments, L 3 comprises , wherein z is an integer between 1 and 5, inclusive.
  • L 3 is , wherein z is an integer between 1 and 10, inclusive. In some embodiments, L 3 is , wherein z is an integer between 1 and 5, inclusive. [0310] In some embodiments, L 3 comprises optionally substituted heteroalkenylene. In some embodiments, L 3 comprises optionally substituted C 1 -C 12 heteroalkenylene. In some embodiments, L 3 comprises optionally substituted C 1 -C 6 heteroalkenylene. In some embodiments, L 3 comprises optionally substituted C 1 -C 3 heteroalkenylene. In some embodiments, L 3 comprises substituted heteroalkenylene. In some embodiments, L 3 comprises substituted C 1 -C 12 heteroalkenylene.
  • L 3 comprises substituted C 1 -C 6 heteroalkenylene. In some embodiments, L 3 comprises substituted C 1 -C 3 heteroalkenylene. In some embodiments, L 3 comprises unsubstituted heteroalkenylene. In some embodiments, L 3 comprises unsubstituted C1-C12 heteroalkenylene. In some embodiments, L 3 comprises unsubstituted C 1 -C 6 heteroalkenylene. In some embodiments, L 3 comprises unsubstituted C 1 -C 3 heteroalkenylene. [0311] In some embodiments, L 3 comprises optionally substituted heteroalkynylene. In some embodiments, L 3 comprises optionally substituted C1-C12 heteroalkynylene.
  • L 3 comprises optionally substituted C1-C6 heteroalkynylene. In some embodiments, L 3 comprises optionally substituted C1-C3 heteroalkynylene. In some embodiments, L 3 comprises substituted heteroalkynylene. In some embodiments, L 3 comprises substituted C1-C12 heteroalkynylene. In some embodiments, L 3 comprises substituted C1-C6 heteroalkynylene. In some embodiments, L 3 comprises substituted C1-C3 heteroalkynylene. In some embodiments, L 3 comprises unsubstituted heteroalkynylene. In some embodiments, L 3 comprises unsubstituted C1-C12 heteroalkynylene.
  • L 3 comprises unsubstituted C1-C6 heteroalkynylene. In some embodiments, L 3 comprises unsubstituted C1-C3 heteroalkynylene. [0312] In some embodiments, L 3 comprises optionally substituted carbocyclylene. In some embodiments, L 3 comprises optionally substituted C3-C10 carbocyclylene. In some embodiments, L 3 comprises optionally substituted C3-C6 carbocyclylene. In some embodiments, L 3 comprises substituted
  • L 3 comprises substituted C 3 -C 10 carbocyclylene. In some embodiments, L 3 comprises substituted C3-C6 carbocyclylene. In some embodiments, L 3 comprises unsubstituted carbocyclylene. In some embodiments, L 3 comprises unsubstituted C3-C10 carbocyclylene. In some embodiments, L 3 comprises unsubstituted C 3 -C 6 carbocyclylene. [0313] In some embodiments, L 3 comprises optionally substituted heterocyclylene. In some embodiments, L 3 comprises optionally substituted 3-10 membered heterocyclylene.
  • L 3 comprises optionally substituted 3-6 membered heterocyclylene. In some embodiments, L 3 comprises substituted heterocyclylene. In some embodiments, L 3 comprises substituted 3-10 membered heterocyclylene. In some embodiments, L 3 comprises substituted 3-6 membered heterocyclylene. In some embodiments, L 3 comprises unsubstituted heterocyclylene. In some embodiments, L 3 comprises unsubstituted 3-10 membered heterocyclylene. In some embodiments, L 3 comprises unsubstituted 3-6 membered heterocyclylene. [0314] In some embodiments, L 3 comprises optionally substituted arylene. In some embodiments, L 3 comprises optionally substituted phenylene.
  • L 3 comprises unsubstituted heteroarylene. In some embodiments, L 3 comprises unsubstituted 5-10 membered heteroarylene. In some embodiments, L 3 comprises unsubstituted 5-6 membered monocyclic heteroarylene.
  • the compound of Formula (IX) is of Formula (IX-B): or a salt thereof, wherein z is an integer between 1 and 100, inclusive. In some embodiments of Formula (IX-B), z is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments of Formula (IX-B), z is 1, 2, 3, 4, or 5. In some embodiments of Formula (IX-B), z is 3. [0317] In some embodiments, the compound of Formula (IX) is of Formula (IX-C):
  • R 4 is optionally substituted C1-12 alkyl. As generally described herein, R 4 is a nitrogen protecting group.
  • the compound of Formula (IX) is of Formula (IX-D): or a salt thereof, wherein z is an integer between 1 and 100, inclusive. In some embodiments of Formula (IX-D), z is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments of Formula (IX-D), z is 1, 2, 3, 4, or 5. In some embodiments of Formula (IX-D), z is 3.
  • the compound of Formula (IX) is of formula: , or a salt thereof. [0321] In some embodiments, R C is -OH.
  • R C is an amino acid moiety or a peptide
  • R N is hydrogen. In some embodiments, R C is an amino acid moiety, and R N is hydrogen. In some embodiments, R C is a peptide, and R N is hydrogen. In some embodiments, R C is an amino acid moiety or a peptide; and R N is an amino acid moiety or a peptide. In some embodiments, R C is an amino acid moiety; and R N is an amino acid moiety. In some embodiments, R C is an amino acid moiety; and R N is a peptide. In some embodiments, R C is a peptide; and R N is an amino acid moiety. In some embodiments, R C is a peptide; and R N is an amino acid moiety. In some embodiments, R C is a peptide; and R N is a peptide. [0322] In some embodiments, the compound of Formula (VIII) is of Formula (VIII-A):
  • z is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.
  • z is 1, 2, 3, 4, or 5.
  • z is 3.
  • R C is an amino acid moiety or a peptide.
  • R N is an amino acid moiety or a peptide.
  • R C is an amino acid moiety or a peptide; and R N is an amino acid moiety or a peptide.
  • R C is an amino acid moiety or a peptide; R N is an amino acid moiety or a peptide; and z is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.
  • R C is an amino acid moiety or a peptide; R N is an amino acid moiety or a peptide; and z is 1, 2, 3, 4, or 5.
  • R C is an amino acid moiety or a peptide
  • R N is an amino acid moiety or a peptide
  • z is 3. Additional Embodiments of Methods of Preparation of Formulae (III-i), (VI), and (VIII) [0323]
  • the compound of Formula (V), or salt thereof is a peptide comprising a cysteine residue.
  • the peptide comprises a C-terminal cysteine residue.
  • the peptide comprises an internal cysteine residue.
  • the peptide is somatostatin 14 (SST-14), somatostatin 28 (SST-28), hepatitis B virus core antigen protein (HBcAg), vimentin, glial fibrillary acidic protein (GFAP), desmin, human serum albumin (HSA), interleukin-1 receptor antagonist protein (IL1RA), rabies virus glycoprotein (RVG), or Tau peptide (306-336).
  • the peptide is somatostatin 14 (SST-14).
  • the peptide is somatostatin 28 (SST-28).
  • the peptide is hepatitis B virus core antigen protein (HBcAg).
  • the peptide is vimentin. In some embodiments, the peptide is glial fibrillary acidic protein (GFAP). In some embodiments, the peptide is desmin. In some embodiments, the peptide is human serum albumin (HSA). In some embodiments, the peptide is interleukin-1 receptor antagonist protein (IL1RA). In some embodiments, the peptide is rabies virus glycoprotein (RVG). In some embodiments, the peptide is Tau peptide (306-336). [0325] In some embodiments, the peptide is QP1094, Tet-20, QP300, QP1112, or QP1149. In some embodiments, the peptide is QP1094. In some embodiments, the peptide is Tet-20.
  • the peptide is QP300. In some embodiments, the peptide is QP1112. In some embodiments, the peptide is QP1149. [0326] In some embodiments, the method is performed at a pH of between about 6.0 and about 10.0, about 6.5 and about 9.5, about 6.5 and about 9.0, about 6.5 and about 8.5, about 7.0 and about 9.0, about 7.0 and about 8.5, about 7.0 and about 8.0, or about 7.4 and about 8.0. In some embodiments, the method is performed at a pH of between about 7.0 and about 9.0. In some embodiments, the method is performed at a pH of between about 7.0 and about 8.0.
  • the method is performed at a pH of between about 7.4 and about 8.0. In some embodiments, the method is performed at a pH of between about 7.0 and about 9.0, preferably wherein the method is performed at a pH of about 7.4 or about 8.0. [0327] In some embodiments, the method is performed at a pH of about 6.5, about 6.6, about 6.7, about 6.8, about 6.9, about 7.0, about 7.1, about 7.2, about 7.3, about 7.4, about 7.5, about 7.6, about 7.7, about 7.8, about 7.9, about 8.0, about 8.1, about 8.2, about 8.3, about 8.4, or about 8.5. In some embodiments,
  • the method is performed at a temperature of between about 35 °C and about 40 °C. In some embodiments, the method is performed at a temperature of between about 20 °C and about 50 °C, preferably wherein the method is performed at a temperature of about 37 °C.
  • the method is performed at a temperature of 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 41 °C, about 42 °C, about 43 °C, about 44 °C, about 45 °C, about 46 °C, about 47 °C, about 48 °C, about 49 °C, or about 50 °C.
  • the method is performed at a temperature of about 37 °C.
  • the method further comprises one or more steps selected from: contacting a reaction mixture comprising the product with a polymer particle comprising a thiol group or a dibenzocyclooctynyl group; subjecting the reaction mixture to dialysis; and subjecting the reaction mixture to a desalting column.
  • the method further comprises contacting a reaction mixture comprising the product with a polymer particle comprising a thiol group or a dibenzocyclooctynyl group.
  • the method further comprises subjecting the reaction mixture to dialysis.
  • the method further comprises subjecting the reaction mixture to a desalting column.
  • the method further comprises enzymatic digestion of the peptide or the compound of any one of Formula (III-i), Formula (VI), or Formula (VIII), or salt thereof.
  • enzymatic digestion of the peptide or the compound of any one of Formula (III-i), Formula (VI), or Formula (VIII), or salt thereof comprises exposing the peptide or the compound of any one of Formula (III-i), Formula (VI), or Formula (VIII), or salt thereof, to a polypeptide-cleaving agent.
  • the method further comprises enzymatic digestion of the peptide or the compound of Formula (III-i), or salt thereof.
  • enzymatic digestion of the peptide or the compound of Formula (III-i), or salt thereof comprises exposing the peptide or the compound of Formula (III-i), or salt thereof, to a polypeptide-cleaving agent.
  • the method further comprises enzymatic digestion of the peptide or the compound of Formula (VI), or salt thereof.
  • enzymatic digestion of the peptide or the compound of Formula (VI), or salt thereof comprises exposing the peptide or the compound of Formula (VI), or salt thereof, to a polypeptide- cleaving agent.
  • the method further comprises enzymatic digestion of the peptide or the compound of Formula (VIII), or salt thereof.
  • enzymatic digestion of the peptide or the compound of Formula (VIII), or salt thereof comprises exposing the peptide or the
  • the polypeptide-cleaving agent comprises a protease.
  • the protease comprises Arg-C, Lys-C, Glu-C, trypsin, chymotrypsin, and/or Asp-N.
  • the protease comprises Arg-C.
  • the protease comprises Lys-C.
  • the protease comprises Glu-C.
  • the protease comprises trypsin.
  • the protease comprises chymotrypsin. In some embodiments, the protease comprises Asp-N. In some embodiments, the protease comprises Lys-C, Glu-C, and/or Asp-N. [0333] In some embodiments, the peptide is a product of Lys-C digest of HSA, a product of Glu-C digest of HSA, a product of Glu-C digest of GFAP, a product of Asp-N digest of GFAP, or a product of Glu-C digest of vimentin. In some embodiments, the peptide is a product of Lys-C digest of HSA.
  • the peptide is a product of Glu-C digest of HSA. In some embodiments, the peptide is a product of Glu-C digest of GFAP. In some embodiments, the peptide is a product of Asp-N digest of GFAP. In some embodiments, the peptide is or a product of Glu-C digest of vimentin. [0334] In some embodiments, the method further comprises contacting the peptide or the compound of any one of Formula (III-i), Formula (VI), or Formula (VIII), or salt thereof, with tris(2- carboxyethyl)phosphine (TCEP).
  • TCEP tris(2- carboxyethyl)phosphine
  • the method further comprises contacting the peptide or the compound of Formula (III-i), or salt thereof, with tris(2-carboxyethyl)phosphine (TCEP). In some embodiments, the method further comprises contacting the peptide or the compound of Formula (VI), or salt thereof, with tris(2-carboxyethyl)phosphine (TCEP). In some embodiments, the method further comprises contacting the peptide or the compound of Formula (VIII), or salt thereof, with tris(2- carboxyethyl)phosphine (TCEP).
  • the method further comprises contacting the compound of any one of Formula (III-i), Formula (VI), or Formula (VIII), or salt thereof, with a compound comprising a complementary moiety, such that the azide moiety of the compound of any one of Formula (III-i), Formula (VI), or Formula (VIII), or salt thereof, reacts with the complementary moiety via click chemistry to form a triazole-linked conjugated peptide.
  • the method further comprises contacting the compound of Formula (III-i), or salt thereof, with a compound comprising a complementary moiety, such that the azide moiety of the compound of Formula (III-i), or salt thereof, reacts with the complementary moiety via click chemistry to form a triazole-linked conjugated peptide.
  • the method further comprises contacting the compound of Formula (VI), or salt thereof, with a compound comprising a complementary moiety, such that the azide moiety of the compound of Formula (VI), or salt thereof, reacts with the complementary moiety via click chemistry to form a triazole-linked conjugated peptide.
  • the method further comprises contacting the compound of Formula (VIII), or salt thereof, with a compound comprising a complementary moiety, such that the azide moiety of the compound of Formula (VIII), or salt thereof, reacts with the complementary moiety via click chemistry to form a triazole-linked conjugated peptide.
  • the compound comprising a complementary moiety is of Formula (X):
  • each of R 3a and R 3b is independently hydrogen, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl, or R 3a and R 3b are joined together with their intervening atoms to form optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl.
  • at least one of R 3a and R 3b is hydrogen.
  • R 3a is hydrogen.
  • R 3b is hydrogen. [0338] In some embodiments, at least one of R 3a and R 3b is optionally substituted aliphatic. In some embodiments, R 3a is optionally substituted aliphatic. In some embodiments, R 3b is optionally substituted aliphatic. In some embodiments, at least one of R 3a and R 3b is substituted aliphatic. In some embodiments, R 3a is substituted aliphatic. In some embodiments, R 3b is substituted aliphatic. [0339] In some embodiments, at least one of R 3a and R 3b is optionally substituted alkyl. In some embodiments, at least one of R 3a and R 3b is optionally substituted C1-C12 alkyl.
  • At least one of R 3a and R 3b is optionally substituted C1-C6 alkyl. In some embodiments, at least one of R 3a and R 3b is optionally substituted C1-C3 alkyl. In some embodiments, at least one of R 3a and R 3b is optionally substituted linear alkyl. In some embodiments, at least one of R 3a and R 3b is substituted alkyl. In some embodiments, at least one of R 3a and R 3b is substituted C1-C12 alkyl. In some embodiments, at least one of R 3a and R 3b is substituted C1-C6 alkyl.
  • At least one of R 3a and R 3b is substituted C1-C3 alkyl. In some embodiments, at least one of R 3a and R 3b is substituted linear alkyl. In some embodiments, R 3a is optionally substituted alkyl. In some embodiments, R 3b is optionally substituted alkyl. [0340] In some embodiments, at least one of R 3a and R 3b is optionally substituted alkenyl. In some embodiments, at least one of R 3a and R 3b is optionally substituted C1-C12 alkenyl. In some embodiments, at least one of R 3a and R 3b is optionally substituted C 1 -C 6 alkenyl.
  • At least one of R 3a and R 3b is optionally substituted C 1 -C 3 alkenyl. In some embodiments, at least one of R 3a and R 3b is substituted alkenyl. In some embodiments, at least one of R 3a and R 3b is substituted C 1 -C 12 alkenyl. In some embodiments, at least one of R 3a and R 3b is substituted C 1 -C 6 alkenyl. In some embodiments, at least one of R 3a and R 3b is substituted C 1 -C 3 alkenyl. In some embodiments, R 3a is optionally substituted alkenyl. In some embodiments, R 3b is optionally substituted alkenyl.
  • R0708.70175WO00 92/180 #13640937v1 [0342] In some embodiments, at least one of R 3a and R 3b is optionally substituted heteroaliphatic. In some embodiments, R 3a is optionally substituted heteroaliphatic. In some embodiments, R 3b is optionally substituted heteroaliphatic. In some embodiments, at least one of R 3a and R 3b is substituted heteroaliphatic. In some embodiments, R 3a is substituted heteroaliphatic. In some embodiments, R 3b is substituted heteroaliphatic. [0343] In some embodiments, at least one of R 3a and R 3b is optionally substituted heteroalkyl.
  • At least one of R 3a and R 3b is optionally substituted C1-C6 heteroalkenyl. In some embodiments, at least one of R 3a and R 3b is optionally substituted C1-C3 heteroalkenyl. In some embodiments, at least one of R 3a and R 3b is substituted heteroalkenyl. In some embodiments, at least one of R 3a and R 3b is substituted C1-C12 heteroalkenyl. In some embodiments, at least one of R 3a and R 3b is substituted C1-C6 heteroalkenyl. In some embodiments, at least one of R 3a and R 3b is substituted C1-C3 heteroalkenyl.
  • R 3a is optionally substituted heteroalkenyl. In some embodiments, R 3b is optionally substituted heteroalkenyl. [0345] In some embodiments, at least one of R 3a and R 3b is optionally substituted heteroalkynyl. In some embodiments, at least one of R 3a and R 3b is optionally substituted C 1 -C 12 heteroalkynyl. In some embodiments, at least one of R 3a and R 3b is optionally substituted C 1 -C 6 heteroalkynyl. In some embodiments, at least one of R 3a and R 3b is optionally substituted C 1 -C 3 heteroalkynyl.
  • At least one of R 3a and R 3b is optionally substituted carbocyclyl. In some embodiments, at least one of R 3a and R 3b is optionally substituted C 3 -C 10 carbocyclyl. In some embodiments, at least one of R 3a and R 3b is optionally substituted C 3 -C 6 carbocyclyl. In some embodiments, at least one of R 3a and R 3b is substituted carbocyclyl. In some embodiments, at least one of
  • At least one of R 3a and R 3b is optionally substituted 3-6 membered heterocyclyl. In some embodiments, at least one of R 3a and R 3b is substituted heterocyclyl. In some embodiments, at least one of R 3a and R 3b is substituted 3-10 membered heterocyclyl. In some embodiments, at least one of R 3a and R 3b is substituted 3-6 membered heterocyclyl. In some embodiments, R 3a is optionally substituted heterocyclyl. In some embodiments, R 3b is optionally substituted heterocyclyl. [0348] In some embodiments, at least one of R 3a and R 3b is optionally substituted aryl.
  • At least one of R 3a and R 3b is optionally substituted phenyl. In some embodiments, at least one of R 3a and R 3b is substituted aryl. In some embodiments, at least one of R 3a and R 3b is substituted phenyl. In some embodiments, R 3a is optionally substituted aryl. In some embodiments, R 3b is optionally substituted aryl. [0349] In some embodiments, at least one of R 3a and R 3b is optionally substituted heteroaryl. In some embodiments, at least one of R 3a and R 3b is optionally substituted 5-10 membered heteroaryl.
  • At least one of R 3a and R 3b is optionally substituted 5-6 membered monocyclic heteroaryl. In some embodiments, at least one of R 3a and R 3b is substituted heteroaryl. In some embodiments, at least one of R 3a and R 3b is substituted 5-10 membered heteroaryl. In some embodiments, at least one of R 3a and R 3b is substituted 5-6 membered monocyclic heteroaryl. In some embodiments, R 3a is optionally substituted heteroaryl. In some embodiments, R 3b is optionally substituted heteroaryl.
  • R 3a and R 3b are joined together with their intervening atoms to form optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl. In some embodiments, R 3a and R 3b are joined together with their intervening atoms to form substituted carbocyclyl, substituted heterocyclyl, substituted aryl, or substituted heteroaryl. In some embodiments, R 3a and R 3b are joined together with their intervening atoms to form optionally substituted carbocyclyl or optionally substituted heterocyclyl.
  • R 3a and R 3b are joined together with their intervening atoms to form substituted carbocyclyl or substituted heterocyclyl. [0351] In some embodiments, R 3a and R 3b are joined together with their intervening atoms to form optionally substituted carbocyclyl. In some embodiments, R 3a and R 3b are joined together with their intervening atoms to form optionally substituted C 3 -C 10 carbocyclyl. In some embodiments, R 3a and R 3b are joined together with their intervening atoms to form optionally substituted unsaturated C 3 -C 10 carbocyclyl.
  • R 3a and R 3b are joined together with their intervening atoms to form optionally substituted cyclooctenyl. In some embodiments, R 3a and R 3b are joined together with their intervening atoms to form substituted carbocyclyl. In some embodiments, R 3a and R 3b are joined together with their intervening atoms to form substituted C 3 -C 10 carbocyclyl. In some embodiments, R 3a and R 3b
  • R0708.70175WO00 94/180 #13640937v1 are joined together with their intervening atoms to form substituted unsaturated C 3 -C 10 carbocyclyl.
  • R 3a and R 3b are joined together with their intervening atoms to form substituted cyclooctenyl.
  • R 3a and R 3b are joined together with their intervening atoms to form optionally substituted heterocyclyl.
  • R 3a and R 3b are joined together with their intervening atoms to form optionally substituted 3-10 membered heterocyclyl.
  • R 3a and R 3b are joined together with their intervening atoms to form optionally substituted unsaturated 3-10 membered heterocyclyl. In some embodiments, R 3a and R 3b are joined together with their intervening atoms to form optionally substituted unsaturated 3-10 membered heterocyclyl containing 1 ring N atom. In some embodiments, R 3a and R 3b are joined together with their intervening atoms to form substituted heterocyclyl. In some embodiments, R 3a and R 3b are joined together with their intervening atoms to form substituted 3-10 membered heterocyclyl.
  • R 3a and R 3b are joined together with their intervening atoms to form substituted unsaturated 3-10 membered heterocyclyl. In some embodiments, R 3a and R 3b are joined together with their intervening atoms to form substituted unsaturated 3-10 membered heterocyclyl containing 1 ring N atom. [0353] In some embodiments, R 3a and R 3b are joined together with their intervening atoms to form optionally substituted aryl. In some embodiments, R 3a and R 3b are joined together with their intervening atoms to form optionally substituted phenyl. In some embodiments, R 3a and R 3b are joined together with their intervening atoms to form substituted aryl.
  • R 3a and R 3b are joined together with their intervening atoms to form substituted phenyl. In some embodiments, R 3a and R 3b are joined together with their intervening atoms to form optionally substituted heteroaryl. In some embodiments, R 3a and R 3b are joined together with their intervening atoms to form optionally substituted 5-10 membered heteroaryl. In some embodiments, R 3a and R 3b are joined together with their intervening atoms to form substituted heteroaryl. In some embodiments, R 3a and R 3b are joined together with their intervening atoms to form substituted 5-10 membered heteroaryl. [0354] In some embodiments, the compound of Formula (X) is of formula: , wherein R 3C is optionally substituted aliphatic, optionally substituted heteroaliphatic,
  • R0708.70175WO00 95/180 #13640937v1 optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, or –OR A .
  • R 3C is optionally substituted aliphatic.
  • R 3C is optionally substituted heteroaliphatic.
  • R 3C is optionally substituted carbocyclyl.
  • R 3C is optionally substituted heterocyclyl.
  • R 3C is optionally substituted aryl.
  • R 3C is optionally substituted heteroaryl.
  • R 3C is –OR A .
  • the compound comprising a complementary moiety comprises a strained alkyne. In some embodiments, the compound comprising a complementary moiety comprises a cyclooctyne or an azacyclooctyne. In some embodiments, the compound comprising a complementary moiety comprises a cyclooctyne. In some embodiments, the compound comprising a complementary moiety comprises an azacyclooctyne.
  • the compound comprising a complementary moiety comprises dibenzoazacyclooctyne (DIBAC or DBCO), biarylazacyclooctynone (BARAC), dibenzocyclooctyne (DIBO), difluorinated cyclooctyne (DIFO), bicyclononyne (BCN), dimethoxyazacyclooctyne (DIMAC), monofluorinated cyclooctyne (MOFO), cyclooctyne (OCT), and/or aryl-less cyclooctyne (ALO).
  • DIBAC or DBCO dibenzoazacyclooctyne
  • BARAC dibenzocyclooctynone
  • DIBO dibenzocyclooctyne
  • DIFO difluorinated cyclooctyne
  • BCN bicyclononyne
  • DIMAC dimethoxyazacycloo
  • the compound comprising a complementary moiety comprises dibenzoazacyclooctyne (DIBAC or DBCO). In some embodiments, the compound comprising a complementary moiety comprises biarylazacyclooctynone (BARAC). In some embodiments, the compound comprising a complementary moiety comprises dibenzocyclooctyne (DIBO). In some embodiments, the compound comprising a complementary moiety comprises difluorinated cyclooctyne (DIFO). In some embodiments, the compound comprising a complementary moiety comprises bicyclononyne (BCN). In some embodiments, the compound comprising a complementary moiety comprises dimethoxyazacyclooctyne (DIMAC).
  • DIBAC dibenzoazacyclooctyne
  • DBCO dibenzoazacyclooctyne
  • the compound comprising a complementary moiety comprises biarylazacyclooctynone (BARAC).
  • the compound comprising a complementary moiety comprises monofluorinated cyclooctyne (MOFO). In some embodiments, the compound comprising a complementary moiety comprises cyclooctyne (OCT). In some embodiments, the compound comprising a complementary moiety comprises aryl-less cyclooctyne (ALO). [0356] In some embodiments, the compound comprising a complementary moiety comprises a linking group. In some embodiments, the linking group comprises a polypeptidyl group. [0357] In some embodiments, the linking group comprises a polypeptidyl group.
  • the polypeptidyl group comprises at least 5 amino acid residues, at least 10 amino acid residues, at least 15 amino acid residues, or at least 20 amino acid residues. In certain embodiments, the polypeptidyl group comprises between 5 and 10 amino acid residues, between 5 and 15 amino acid residues, between 5 and 20 amino acid residues, between 10 and 15 amino acid residues, between 10 and 20 amino acid residues, or between 15 and 20 amino acid residues. In some embodiments, the polypeptidyl group comprises between 5 and 15 amino acid residues.
  • the polypeptidyl group has a length of at least about 20 ⁇ , 25 ⁇ , 30 ⁇ , 35 ⁇ , 40 ⁇ , 45 ⁇ , 50 ⁇ , 55 ⁇ , 60 ⁇ , 65 ⁇ , 70 ⁇ , or 75 ⁇ .
  • the polypeptidyl group has a length in a range from 20 ⁇ to 30 ⁇ , 20 ⁇ to 35 ⁇ , 20 ⁇ to 40 ⁇ , 20 ⁇ to 45 ⁇ , 20 ⁇ to 50 ⁇ , 20 ⁇ to 55 ⁇ , 20 ⁇ to 60 ⁇ , 20 ⁇ to 65 ⁇ , 20 ⁇ to 70 ⁇ , 20 ⁇ to 75 ⁇ , 30 ⁇ to 40 ⁇ , 30 ⁇ to 45 ⁇ , 30 ⁇ to 50
  • the polypeptidyl group comprises at least 1 negatively charged moiety at physiological pH.
  • the polypeptidyl group comprises at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, or at least 15 negatively charged moieties at physiological pH.
  • the polypeptidyl group comprises between 1 and 2, 1 and 3, 1 and 4, 1 and 5, 1 and 6, 1 and 7, 1 and 8, 1 and 9, 1 and 10, 1 and 11, 1 and 12, 1 and 13, 1 and 14, 1 and 15, 2 and 3, 2 and 4, 2 and 5, 2 and 6, 2 and 7, 2 and 8, 2 and 9, 2 and 10, 2 and 11, 2 and 12, 2 and 13, 2 and 14, 2 and 15, 3 and 4, 3 and 5, 3 and 6, 3 and 7, 3 and 8, 3 and 9, 3 and 10, 3 and 11, 3 and 12, 3 and 13, 3 and 14, 3 and 15, 4 and 5, 4 and 6, 4 and 7, 4 and 8, 4 and 9, 4 and 10, 4 and 11, 4 and 12, 4 and 13, 4 and 14, 4 and 15, 5 and 6, 5 and 7, 5 and 8, 5 and 9, 5 and 10, 5 and 11, 5 and 12, 5 and 13, 5 and 14, 5 and 15, 6 and 10, 6 and 15, 7 and 10, 7 and 15, 8 and 10, 8 and 15, 9 and 10, 9 and 15, or 10 and 15 negatively charged moieties at physiological pH.
  • the polypeptidyl group comprises between 1 and 10 negatively charged moieties at physiological pH. [0360] In some embodiments, the polypeptidyl group comprises at least 1 aspartate residue. In certain embodiments, the polypeptidyl group comprises at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, or at least 15 aspartate residues.
  • the polypeptidyl group comprises between 1 and 2, 1 and 3, 1 and 4, 1 and 5, 1 and 6, 1 and 7, 1 and 8, 1 and 9, 1 and 10, 1 and 11, 1 and 12, 1 and 13, 1 and 14, 1 and 15, 2 and 3, 2 and 4, 2 and 5, 2 and 6, 2 and 7, 2 and 8, 2 and 9, 2 and 10, 2 and 11, 2 and 12, 2 and 13, 2 and 14, 2 and 15, 3 and 4, 3 and 5, 3 and 6, 3 and 7, 3 and 8, 3 and 9, 3 and 10, 3 and 11, 3 and 12, 3 and 13, 3 and 14, 3 and 15, 4 and 5, 4 and 6, 4 and 7, 4 and 8, 4 and 9, 4 and 10, 4 and 11, 4 and 12, 4 and 13, 4 and 14, 4 and 15, 5 and 6, 5 and 7, 5 and 8, 5 and 9, 5 and 10, 5 and 11, 5 and 12, 5 and 13, 5 and 14, 5 and 15, 6 and 10, 6 and 15, 7 and 10, 7 and 15, 8 and 10, 8 and 15, 9 and 10, 9 and 15, or 10 and 15 aspartate residues.
  • the polypeptidyl group comprises between 1 and 10 aspartate residues. [0361] In some embodiments, the polypeptidyl group comprises at least 1 phenylalanine residue. In certain embodiments, the polypeptidyl group comprises at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, or at least 15 phenylalanine residues.
  • the polypeptidyl group comprises between 1 and 2, 1 and 3, 1 and 4, 1 and 5, 1 and 6, 1 and 7, 1 and 8, 1 and 9, 1 and 10, 1 and 11, 1 and 12, 1 and 13, 1 and 14, 1 and 15, 2 and 3, 2 and 4, 2 and 5, 2 and 6, 2 and 7, 2 and 8, 2 and 9, 2 and 10, 2 and 11, 2 and 12, 2 and 13, 2 and 14, 2 and 15, 3 and 4, 3 and 5, 3 and 6, 3 and 7, 3 and 8, 3 and 9, 3 and 10, 3 and 11, 3 and 12, 3 and 13, 3 and 14, 3 and 15, 4 and 5, 4 and 6, 4 and 7, 4 and 8, 4 and 9, 4 and 10, 4 and 11, 4 and 12, 4 and 13, 4 and 14, 4 and 15, 5 and 6, 5 and 7, 5 and 8, 5 and 9, 5 and 10, 5 and 11, 5 and 12, 5 and 13, 5 and 14, 5 and 15, 6 and 10, 6 and 15, 7 and 10, 7 and 15, 8 and 10, 8 and 15, 9 and 10, 9 and 15, or
  • the polypeptidyl group comprises at least 1 glycine residue. In certain embodiments, the polypeptidyl group comprises at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, or at least 15 glycine residues.
  • the polypeptidyl group comprises between 1 and 2, 1 and 3, 1 and 4, 1 and 5, 1 and 6, 1 and 7, 1 and 8, 1 and 9, 1 and 10, 1 and 11, 1 and 12, 1 and 13, 1 and 14, 1 and 15, 2 and 3, 2 and 4, 2 and 5, 2 and 6, 2 and 7, 2 and 8, 2 and 9, 2 and 10, 2 and 11, 2 and 12, 2 and 13, 2 and 14, 2 and 15, 3 and 4, 3 and 5, 3 and 6, 3 and 7, 3 and 8, 3 and 9, 3 and 10, 3 and 11, 3 and 12, 3 and 13, 3 and 14, 3 and 15, 4 and 5, 4 and 6, 4 and 7, 4 and 8, 4 and 9, 4 and 10, 4 and 11, 4 and 12, 4 and 13, 4 and 14, 4 and 15, 5 and 6, 5 and 7, 5 and 8, 5 and 9, 5 and 10, 5 and 11, 5 and 12, 5 and 13, 5 and 14, 5 and 15, 6 and 10, 6 and 15, 7 and 10, 7 and 15, 8 and 10, 8 and 15, 9 and 10, 9 and 15, or 10 and 15 glycine residues.
  • the polypeptidyl group comprises at least 1 proline residue. In certain embodiments, the polypeptidyl group comprises at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, or at least 15 proline residues.
  • the polypeptidyl group comprises between 1 and 2, 1 and 3, 1 and 4, 1 and 5, 1 and 6, 1 and 7, 1 and 8, 1 and 9, 1 and 10, 1 and 11, 1 and 12, 1 and 13, 1 and 14, 1 and 15, 2 and 3, 2 and 4, 2 and 5, 2 and 6, 2 and 7, 2 and 8, 2 and 9, 2 and 10, 2 and 11, 2 and 12, 2 and 13, 2 and 14, 2 and 15, 3 and 4, 3 and 5, 3 and 6, 3 and 7, 3 and 8, 3 and 9, 3 and 10, 3 and 11, 3 and 12, 3 and 13, 3 and 14, 3 and 15, 4 and 5, 4 and 6, 4 and 7, 4 and 8, 4 and 9, 4 and 10, 4 and 11, 4 and 12, 4 and 13, 4 and 14, 4 and 15, 5 and 6, 5 and 7, 5 and 8, 5 and 9, 5 and 10, 5 and 11, 5 and 12, 5 and 13, 5 and 14, 5 and 15, 6 and 10, 6 and 15, 7 and 10, 7 and 15, 8 and 10, 8 and 15, 9 and 10, 9 and 15, or 10 and 15 proline residues.
  • the polypeptidyl group comprises at least 1 DD repeat, GG repeat, FF repeat, DDD repeat, GGG, and/or FFF repeat. In certain embodiments, the polypeptidyl group comprises at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, or at least 15 DD repeats, GG repeats, FF repeats, DDD repeats, GGG, and/or FFF repeats.
  • the polypeptidyl group comprises between 1 and 2, 1 and 3, 1 and 4, 1 and 5, 1 and 6, 1 and 7, 1 and 8, 1 and 9, 1 and 10, 1 and 11, 1 and 12, 1 and 13, 1 and 14, 1 and 15, 2 and 3, 2 and 4, 2 and 5, 2 and 6, 2 and 7, 2 and 8, 2 and 9, 2 and 10, 2 and 11, 2 and 12, 2 and 13, 2 and 14, 2 and 15, 3 and 4, 3 and 5, 3 and 6, 3 and 7, 3 and 8, 3 and 9, 3 and 10, 3 and 11, 3 and 12, 3 and 13, 3 and 14, 3 and 15, 4 and 5, 4 and 6, 4 and 7, 4 and 8, 4 and 9, 4 and 10, 4 and 11, 4 and 12, 4 and 13, 4 and 14, 4 and 15, 5 and 6, 5 and 7, 5 and 8, 5 and 9, 5 and 10, 5 and 11, 5 and 12, 5 and 13, 5 and 14, 5 and 15, 6 and 10, 6 and 15, 7 and 10, 7 and 15, 8 and 10, 8 and 15, 9 and 10, 9 and 15, or 10 and 15 DD repeats, GG repeats, FF repeats, DDD repeats, GGG, and/or FFF repeats.
  • the polypeptidyl group comprises a sequence selected from the group consisting of GPPPPPPPPG (SEQ ID NO: 11), isoEGWRW (SEQ ID NO: 12), DDGGGDDDFF (SEQ ID NO: 13), GGSSSGSGNDEEFQ (SEQ ID NO: 14), GGGGGDPDPDFF (SEQ ID NO: 15),
  • the polypeptidyl group comprises DDGGGDDDFF (SEQ ID NO: 13).
  • the linking group comprises an oligonucleotide. In certain embodiments, the oligonucleotide is a single-stranded oligonucleotide.
  • the oligonucleotide is a double-stranded oligonucleotide. In certain embodiments, the oligonucleotide has a length of at least 15, at least 20, at least 25, at least 30, at least 35, at least 40, at least 45, or at least 50 nucleotides.
  • the oligonucleotide has a length in a range from 15 to 20, 15 to 25, 15 to 30, 15 to 35, 15 to 40, 15 to 45, 15 to 50, 20 to 25, 20 to 30, 20 to 35, 20 to 40, 20 to 45, 20 to 50, 25 to 30, 25 to 35, 25 to 40, 25 to 45, 25 to 50, 30 to 35, 30 to 40, 30 to 45, 30 to 50, 35 to 40, 35 to 45, 35 to 50, 40 to 45, 40 to 50, or 45 to 50 nucleotides. In some embodiments, the oligonucleotide has a length of at least 25 nucleotides.
  • At least one strand of the oligonucleotide has a sequence that is at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% identical to 5'-CCACGCGTGGAACCCTTGGGATCCA- 3' (SEQ ID NO: 19). In some embodiments, at least one strand of the oligonucleotide has a sequence that is at least 80% identical to 5'-CCACGCGTGGAACCCTTGGGATCCA-3' (SEQ ID NO: 19).
  • At least one strand of the oligonucleotide has a sequence that is at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% identical to 5'-TGG AGT CAA GGT CCT CTG ATG CCA T-3’ (SEQ ID NO: 20).
  • the linking group further comprises at least one of optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted heteroalkylene, optionally substituted heteroalkenylene, optionally substituted heteroalkynylene, optionally substituted heterocyclylene, optionally substituted carbocyclylene, optionally substituted arylene, optionally substituted heteroarylene, or a combination thereof.
  • the compound comprising a complementary moiety comprises a binding group.
  • the binding group comprises a biotin moiety.
  • the biotin moiety is a bis-biotin moiety.
  • the binding group comprises at least one tag sequence.
  • the at least one tag sequence comprises at least one biotin ligase recognition sequence that permits biotinylation of the compound comprising the complementary moiety (e.g., incorporation of one or more biotin moieties, including biotin and bis-biotin moieties).
  • the at least one tag sequence comprises two biotin ligase recognition sequences oriented in tandem.
  • a biotin ligase recognition sequence refers to an amino acid sequence that is recognized by a biotin ligase, which catalyzes a covalent linkage between the sequence and a biotin molecule.
  • Each biotin ligase recognition sequence of a tag sequence can be covalently linked to a biotin moiety, such that a tag sequence having multiple biotin ligase recognition sequences can be covalently linked to multiple biotin
  • a region of a tag sequence having one or more biotin ligase recognition sequences can be generally referred to as a biotinylation tag or a biotinylation sequence.
  • a bis-biotin or bis-biotin moiety can refer to two biotins bound to two biotin ligase recognition sequences oriented in tandem.
  • the binding group comprises at least one biotin ligase recognition sequence having a biotin moiety attached thereto or at least two biotin ligase recognition sequences, each having a biotin moiety attached thereto.
  • the binding group comprises or is conjugated to an avidin protein.
  • the biotin moiety comprises an avidin protein.
  • the biotin moiety is conjugated to an avidin protein.
  • avidin protein refers to a biotin-binding protein, generally having a biotin binding site at each of four subunits of the avidin protein.
  • avidin proteins include avidin, streptavidin, traptavidin, tamavidin, bradavidin, xenavidin, and homologs and variants thereof.
  • the avidin protein may have a monomeric, dimeric, or tetrameric form.
  • the avidin protein is streptavidin in a tetrameric form (e.g., a homotetramer).
  • the streptavidin in a tetrameric form may be bound to one component (e.g., a first component comprising a first mono-biotin moiety or a first bis-biotin moiety), two components (e.g., a first component comprising a first mono-biotin moiety or a first bis-biotin moiety and a second component comprising a second mono-biotin moiety or a second bis-biotin moiety), three components (e.g., a first component comprising a first bis-biotin moiety, a second component comprising a first mono-biotin moiety, and a third component comprising a second mono-biotin moiety), or four components (e.g., four components, each comprising a mono-biotin moiety).
  • the compound comprising a complementary moiety is immobilized to a surface (e.g., a surface of a sample well). In some embodiments, the compound comprising a complementary moiety is immobilized to a surface of a sample well.
  • a method of functionalizing a peptide having at least one free cysteine residue comprising reacting the peptide with a compound of Formula (II): or a salt thereof, to provide an azide-functionalized peptide, such that the sulfur atom of the cysteine residue in the azide-functionalized peptide is bonded to a group of the formula:
  • R0708.70175WO00 100/180 #13640937v1 wherein: X ⁇ is a counterion; n is 0, 1, 2, 3, or 4; L 1 is optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted heteroalkylene, optionally substituted heteroalkenylene, optionally substituted heteroalkynylene, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, optionally substituted heteroarylene, or a combination thereof; each occurrence of R A is independently hydrogen, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, a nitrogen protecting group when attached to a nitrogen atom, an oxygen protecting group when attached to an oxygen atom, or a sulfur protecting group when attached to a sulfur atom, or two occurrences of
  • X ⁇ is F – , Cl – , Br – , or I – . In some embodiments, X ⁇ is Cl – or Br – . In some embodiments, X ⁇ is Cl – . [0376] In some embodiments, X ⁇ is triflate or a halide ion. In some embodiments, X ⁇ is triflate, F – , Cl – , Br – , or I – . In some embodiments, X ⁇ is triflate, Cl – , or Br – . In some embodiments, X ⁇ is triflate or Cl – . [0377] In some embodiments, L 1 comprises optionally substituted alkylene.
  • L 1 comprises optionally substituted C1-C12 alkylene. In some embodiments, L 1 comprises optionally substituted C1-C6 alkylene. In some embodiments, L 1 comprises optionally substituted C1-C3 alkylene. In some embodiments, L 1 comprises optionally substituted linear alkylene. In some embodiments, L 1 comprises optionally substituted linear C1-C12 alkylene. In some embodiments, L 1 comprises optionally substituted linear C1-C6 alkylene. In some embodiments, L 1 comprises optionally substituted linear C1-C3 alkylene. [0378] In some embodiments, L 1 comprises substituted alkylene. In some embodiments, L 1 comprises substituted C1-C12 alkylene.
  • L 1 comprises substituted C1-C6 alkylene. In some embodiments, L 1 comprises substituted C1-C3 alkylene. In some embodiments, L 1 comprises substituted linear alkylene. In some embodiments, L 1 comprises substituted linear C1-C12 alkylene. In some embodiments, L 1 comprises substituted linear C1-C6 alkylene. In some embodiments, L 1 comprises substituted linear C1-C3 alkylene. [0379] In some embodiments, L 1 comprises unsubstituted alkylene. In some embodiments, L 1 comprises unsubstituted C1-C12 alkylene. In some embodiments, L 1 comprises unsubstituted C1-C6 alkylene.
  • L 1 comprises unsubstituted C1-C3 alkylene. In some embodiments, L 1 comprises unsubstituted linear alkylene. In some embodiments, L 1 comprises unsubstituted linear C1-C12 alkylene. In some embodiments, L 1 comprises unsubstituted linear C1-C6 alkylene. In some embodiments, L 1 comprises unsubstituted linear C1-C3 alkylene. [0380] In some embodiments, L 1 comprises methylene, ethylene, n-propylene, n-butylene, n-pentylene, or n-hexylene. In some embodiments, L 1 comprises methylene, ethylene, or n-propylene.
  • L 1 comprises ethylene. [0381] In some embodiments, L 1 is methylene, ethylene, n-propylene, n-butylene, n-pentylene, or n- hexylene. In some embodiments, L 1 is methylene, ethylene, or n-propylene. In some embodiments, L 1 is ethylene. [0382] In some embodiments, L 1 comprises optionally substituted alkenylene. In some embodiments, L 1 comprises optionally substituted C 1 -C 12 alkenylene. In some embodiments, L 1 comprises optionally substituted C 1 -C 6 alkenylene. In some embodiments, L 1 comprises optionally substituted C 1 -C 3 alkenylene.
  • L 1 comprises substituted alkenylene. In some embodiments, L 1 comprises substituted C 1 -C 12 alkenylene. In some embodiments, L 1 comprises substituted C 1 -C 6 alkenylene. In some embodiments, L 1 comprises substituted C 1 -C 3 alkenylene. In some embodiments, L 1 comprises unsubstituted alkenylene. In some embodiments, L 1 comprises unsubstituted C 1 -C 12 alkenylene. In some embodiments, L 1 comprises unsubstituted C 1 -C 6 alkenylene. In some embodiments, L 1 comprises unsubstituted C 1 -C 3 alkenylene.
  • L 1 comprises optionally substituted alkynylene. In some embodiments, L 1 comprises optionally substituted C1-C12 alkynylene. In some embodiments, L 1 comprises optionally substituted C1-C6 alkynylene. In some embodiments, L 1 comprises optionally substituted C1-C3 alkynylene. In some embodiments, L 1 comprises substituted alkynylene. In some embodiments, L 1 comprises substituted C1-C12 alkynylene. In some embodiments, L 1 comprises substituted C1-C6 alkynylene. In some embodiments, L 1 comprises substituted C1-C3 alkynylene.
  • L 1 comprises unsubstituted alkynylene. In some embodiments, L 1 comprises unsubstituted C1-C12 alkynylene. In some embodiments, L 1 comprises unsubstituted C1-C6 alkynylene. In some embodiments, L 1 comprises unsubstituted C1-C3 alkynylene. [0384] In some embodiments, L 1 comprises optionally substituted heteroalkylene. In some embodiments, L 1 comprises optionally substituted C1-C12 heteroalkylene. In some embodiments, L 1 comprises optionally substituted C1-C6 heteroalkylene. In some embodiments, L 1 comprises optionally substituted C1-C3 heteroalkylene.
  • L 1 comprises optionally substituted linear heteroalkylene. In some embodiments, L 1 comprises optionally substituted linear C1-C12 heteroalkylene. In some embodiments, L 1 comprises optionally substituted linear C1-C6 heteroalkylene. In some embodiments, L 1 comprises optionally substituted linear C1-C3 heteroalkylene. [0385] In some embodiments, L 1 comprises substituted heteroalkylene. In some embodiments, L 1 comprises substituted C1-C12 heteroalkylene. In some embodiments, L 1 comprises substituted C1-C6 heteroalkylene. In some embodiments, L 1 comprises substituted C1-C3 heteroalkylene. In some embodiments, L 1 comprises substituted linear heteroalkylene.
  • L 1 comprises substituted linear C1-C12 heteroalkylene. In some embodiments, L 1 comprises substituted linear C1-C6 heteroalkylene. In some embodiments, L 1 comprises substituted linear C1-C3 heteroalkylene. [0386] In some embodiments, L 1 comprises unsubstituted heteroalkylene. In some embodiments, L 1 comprises unsubstituted C1-C12 heteroalkylene. In some embodiments, L 1 comprises unsubstituted C1-C6 heteroalkylene. In some embodiments, L 1 comprises unsubstituted C 1 -C 3 heteroalkylene. In some embodiments, L 1 comprises unsubstituted linear heteroalkylene.
  • L 1 comprises unsubstituted linear C 1 -C 12 heteroalkylene. In some embodiments, L 1 comprises unsubstituted linear C 1 - C 6 heteroalkylene. In some embodiments, L 1 comprises unsubstituted linear C 1 -C 3 heteroalkylene. [0387] In some embodiments, L 1 comprises or , wherein x is an integer between 1 and 100, inclusive. In some embodiments, L 1 is or , wherein x is an integer between 1 and 100, inclusive.
  • x is an integer between 1 and 90, inclusive; between 1 and 80, inclusive; between 1 and 70, inclusive; between 1 and 60, inclusive; between 1 and 50, inclusive; between 1 and 40, inclusive; between 1 and 30, inclusive; between 1 and 20, inclusive; between 1 and 10, inclusive; or
  • x is an integer between 1 and 10, inclusive. In some embodiments, x is an integer between 1 and 5, inclusive. In some embodiments, x is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, x is 1, 2, 3, 4, 5, 6, 7, 8, or 9. In some embodiments, x is 1, 2, 3, 4, 5, 6, 7, or 8. In some embodiments, x is 1, 2, 3, 4, 5, 6, or 7. In some embodiments, x is 1, 2, 3, 4, 5, or 6. In some embodiments, x is 1, 2, 3, 4, or 5. In some embodiments, x is 1, 2, 3, or 4. In some embodiments, x is 1, 2, or 3. In some embodiments, x is 1 or 2.
  • x is 1. In some embodiments, x is 2. In some embodiments, x is 3. In some embodiments, x is 4. In some embodiments, x is 5. In some embodiments, x is 6. In some embodiments, x is 7. In some embodiments, x is 8. In some embodiments, x is 9. In some embodiments, x is 10. , wherein x is an integer between 1 and 10, inclusive. In some embodiments, L 1 comprises or embodiments, inclusive. [0390] In some embodiments, L 1 comprises optionally substituted heteroalkenylene. In some embodiments, L 1 comprises optionally substituted C1-C12 heteroalkenylene. In some embodiments, L 1 comprises optionally substituted C1-C6 heteroalkenylene.
  • L 1 comprises optionally substituted C1-C3 heteroalkenylene. In some embodiments, L 1 comprises substituted heteroalkenylene. In some embodiments, L 1 comprises substituted C1-C12 heteroalkenylene. In some embodiments, L 1 comprises substituted C1-C6 heteroalkenylene. In some embodiments, L 1 comprises substituted C1-C3 heteroalkenylene. In some embodiments, L 1 comprises unsubstituted heteroalkenylene. In some embodiments, L 1 comprises unsubstituted C1-C12 heteroalkenylene. In some embodiments, L 1 comprises unsubstituted C1-C6 heteroalkenylene.
  • L 1 comprises unsubstituted C1-C3 heteroalkenylene. [0391] In some embodiments, L 1 comprises optionally substituted heteroalkynylene. In some embodiments, L 1 comprises optionally substituted C1-C12 heteroalkynylene. In some embodiments, L 1 comprises optionally substituted C1-C6 heteroalkynylene. In some embodiments, L 1 comprises optionally substituted C 1 -C 3 heteroalkynylene. In some embodiments, L 1 comprises substituted heteroalkynylene. In some embodiments, L 1 comprises substituted C1-C12 heteroalkynylene. In some embodiments, L 1 comprises substituted C1-C6 heteroalkynylene. In some embodiments, L 1 comprises substituted C1-C3
  • L 1 comprises unsubstituted heteroalkynylene. In some embodiments, L 1 comprises unsubstituted C1-C12 heteroalkynylene. In some embodiments, L 1 comprises unsubstituted C1-C6 heteroalkynylene. In some embodiments, L 1 comprises unsubstituted C1-C3 heteroalkynylene. [0392] In some embodiments, L 1 comprises optionally substituted carbocyclylene. In some embodiments, L 1 comprises optionally substituted C3-C10 carbocyclylene.
  • L 1 comprises optionally substituted C3-C6 carbocyclylene. In some embodiments, L 1 comprises substituted carbocyclylene. In some embodiments, L 1 comprises substituted C3-C10 carbocyclylene. In some embodiments, L 1 comprises substituted C3-C6 carbocyclylene. In some embodiments, L 1 comprises unsubstituted carbocyclylene. In some embodiments, L 1 comprises unsubstituted C3-C10 carbocyclylene. In some embodiments, L 1 comprises unsubstituted C3-C6 carbocyclylene. [0393] In some embodiments, L 1 comprises optionally substituted heterocyclylene.
  • L 1 comprises optionally substituted 3-10 membered heterocyclylene. In some embodiments, L 1 comprises optionally substituted 3-6 membered heterocyclylene. In some embodiments, L 1 comprises substituted heterocyclylene. In some embodiments, L 1 comprises substituted 3-10 membered heterocyclylene. In some embodiments, L 1 comprises substituted 3-6 membered heterocyclylene. In some embodiments, L 1 comprises unsubstituted heterocyclylene. In some embodiments, L 1 comprises unsubstituted 3-10 membered heterocyclylene. In some embodiments, L 1 comprises unsubstituted 3-6 membered heterocyclylene. [0394] In some embodiments, L 1 comprises optionally substituted arylene.
  • L 1 comprises optionally substituted phenylene. In some embodiments, L 1 comprises substituted arylene. In some embodiments, L 1 comprises substituted phenylene. In some embodiments, L 1 comprises unsubstituted arylene. In some embodiments, L 1 comprises unsubstituted phenylene. [0395] In some embodiments, L 1 comprises optionally substituted heteroarylene. In some embodiments, L 1 comprises optionally substituted 5-10 membered heteroarylene. In some embodiments, L 1 comprises optionally substituted 5-6 membered monocyclic heteroarylene. In some embodiments, L 1 comprises substituted heteroarylene. In some embodiments, L 1 comprises substituted 5-10 membered heteroarylene.
  • L 1 comprises substituted 5-6 membered monocyclic heteroarylene. In some embodiments, L 1 comprises unsubstituted heteroarylene. In some embodiments, L 1 comprises unsubstituted 5-10 membered heteroarylene. In some embodiments, L 1 comprises unsubstituted 5-6 membered monocyclic heteroarylene. [0396] In some embodiments, at least one occurrence of R 1 is halogen, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, –CN, –OR A , –SCN, –SR A , –
  • n is 0 or 1. In some embodiments, n is 1, 2, 3, or 4. In some embodiments, n is 1, 2, or 3. In some embodiments, n is 0. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4.
  • R C is -OH, and R N is an amino acid moiety. In some embodiments, R C is -OH, and R N is a peptide. In some embodiments, R C is an amino acid moiety or a peptide, and R N is hydrogen. In some embodiments, R C is an amino acid moiety, and R N is hydrogen. In some embodiments, R C is a peptide, and R N is hydrogen. In some embodiments, R C is an amino acid moiety or a peptide; and R N is an amino acid moiety or a peptide. In some embodiments, R C is an amino acid moiety; and R N is an amino acid moiety. In some embodiments, R C is an amino acid moiety; and R N is an amino acid moiety.
  • R C is an amino acid moiety; and R N is a peptide. In some embodiments, R C is a peptide; and R N is an amino acid moiety. In some embodiments, R C is a peptide; and R N is a peptide. [0401] In some embodiments, the compound of Formula (II) is of Formula (II-A) or Formula (II-B): or a salt thereof, such that the sulfur atom of the cysteine residue in the functionalized peptide is bonded to a group of the formula:
  • the compound of Formula (II) is of formula: , or a salt thereof, such that a sulfur atom of a cysteine residue in the azide-functionalized peptide is bonded to a group of the formula: .
  • the method further comprises contacting the azide-functionalized peptide with a compound comprising a complementary moiety, such that the azide moiety of the azide- functionalized peptide reacts with the complementary moiety via click chemistry to provide a triazole- linked conjugated peptide.
  • the method further comprises contacting the azide-functionalized peptide with a compound comprising a complementary moiety, wherein the compound comprising a complementary moiety is of Formula (X): or a salt thereof, to provide a triazole-linked conjugated peptide, such that a sulfur atom of a cysteine residue in the triazole-linked conjugated peptide is bonded to a group of the formula: , wherein: each of R 3a and R 3b is independently hydrogen, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl, or R 3a and R 3b are joined together with their intervening atoms to form optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl.
  • R0708.70175WO00 109/180 #13640937v1 peptide is bonded to a group of the formula: [0406]
  • the sulfur atom of the cysteine residue in the triazole-linked conjugated peptide is bonded to a group of the formula: .
  • a method of sequencing a peptide having at least one free cysteine residue comprising: reacting the peptide with a compound of Formula (VII): or a salt thereof, to provide an azide-functionalized peptide, such that the sulfur atom of a cysteine residue in the azide-functionalized peptide is bonded to a group of the formula: performing a peptide sequencing protocol; wherein: L 2 is optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted heteroalkylene, optionally substituted heteroalkenylene, optionally substituted heteroalkynylene, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, optionally substituted heteroarylene, or a combination thereof.
  • L 2 comprises optionally substituted alkylene. In some embodiments, L 2 comprises optionally substituted C1-C12 alkylene. In some embodiments, L 2 comprises optionally substituted C1-C6 alkylene. In some embodiments, L 2 comprises optionally substituted C1-C3 alkylene. In some embodiments, L 2 comprises optionally substituted linear alkylene. In some embodiments, L 2 comprises optionally substituted linear C1-C12 alkylene. In some embodiments, L 2 comprises optionally substituted linear C1-C6 alkylene. In some embodiments, L 2 comprises optionally substituted linear C1-C3 alkylene.
  • L 2 comprises substituted alkylene. In some embodiments, L 2 comprises substituted C1-C12 alkylene. In some embodiments, L 2 comprises substituted C1-C6 alkylene. In some embodiments, L 2 comprises substituted C1-C3 alkylene. In some embodiments, L 2 comprises substituted linear alkylene. In some embodiments, L 2 comprises substituted linear C 1 -C 12 alkylene. In some embodiments, L 2 comprises substituted linear C1-C6 alkylene. In some embodiments, L 2 comprises substituted linear C1-C3 alkylene. In some embodiments, L 2 comprises unsubstituted alkylene.
  • L 2 comprises unsubstituted C1-C12 alkylene. In some embodiments, L 2 comprises unsubstituted C1-C6 alkylene. In some embodiments, L 2 comprises unsubstituted C1-C3 alkylene. In some embodiments, L 2 comprises unsubstituted linear alkylene. In some embodiments, L 2 comprises unsubstituted linear C1-C12 alkylene. In some embodiments, L 2 comprises unsubstituted linear C1-C6 alkylene. In some embodiments, L 2 comprises unsubstituted linear C1-C3 alkylene.
  • L 2 comprises methylene, ethylene, n-propylene, n-butylene, n-pentylene, or n-hexylene. In some embodiments, L 2 comprises methylene, ethylene, or n-propylene. In some embodiments, L 2 comprises ethylene. In some embodiments, L 2 comprises n-propylene. [0411] In some embodiments, L 2 comprises optionally substituted alkenylene. In some embodiments, L 2 comprises optionally substituted C1-C12 alkenylene. In some embodiments, L 2 comprises optionally substituted C1-C6 alkenylene. In some embodiments, L 2 comprises optionally substituted C1-C3 alkenylene.
  • L 2 comprises substituted alkenylene. In some embodiments, L 2 comprises substituted C1-C12 alkenylene. In some embodiments, L 2 comprises substituted C1-C6 alkenylene. In some embodiments, L 2 comprises substituted C1-C3 alkenylene. In some embodiments, L 2 comprises unsubstituted alkenylene. In some embodiments, L 2 comprises unsubstituted C1-C12 alkenylene. In some embodiments, L 2 comprises unsubstituted C1-C6 alkenylene. In some embodiments, L 2 comprises unsubstituted C1-C3 alkenylene. [0412] In some embodiments, L 2 comprises optionally substituted alkynylene.
  • L 2 comprises optionally substituted C 1 -C 12 alkynylene. In some embodiments, L 2 comprises optionally substituted C 1 -C 6 alkynylene. In some embodiments, L 2 comprises optionally substituted C 1 -C 3 alkynylene. In some embodiments, L 2 comprises substituted alkynylene. In some embodiments, L 2 comprises substituted C 1 -C 12 alkynylene. In some embodiments, L 2 comprises substituted C 1 -C 6 alkynylene. In some embodiments, L 2 comprises substituted C 1 -C 3 alkynylene. In some embodiments, L 2 comprises unsubstituted alkynylene.
  • L 2 comprises unsubstituted C 1 -C 12 alkynylene. In some embodiments, L 2 comprises unsubstituted C 1 -C 6 alkynylene. In some embodiments, L 2 comprises unsubstituted C 1 -C 3 alkynylene. [0413] In some embodiments, L 2 comprises optionally substituted heteroalkylene. In some embodiments, L 2 comprises optionally substituted C 1 -C 12 heteroalkylene. In some embodiments, L 2 comprises optionally substituted C 1 -C 6 heteroalkylene. In some embodiments, L 2 comprises optionally substituted C 1 -C 3 heteroalkylene. In some embodiments, L 2 comprises optionally substituted linear heteroalkylene. In some embodiments, L 2 comprises optionally substituted linear C 1 -C 12 heteroalkylene. In some embodiments, L 2 comprises optionally substituted linear C 1 -C 6 heteroalkylene. In some embodiments, L 2 comprises optionally substituted linear C 1 -C 6 heteroalkylene. In
  • L 2 comprises substituted linear C1-C3 heteroalkylene.
  • L 2 comprises unsubstituted heteroalkylene.
  • L 2 comprises unsubstituted C1-C12 heteroalkylene.
  • L 2 comprises unsubstituted C1-C6 heteroalkylene.
  • L 2 comprises unsubstituted C1-C3 heteroalkylene.
  • L 2 comprises unsubstituted linear heteroalkylene.
  • L 2 comprises unsubstituted linear C1-C12 heteroalkylene.
  • L 2 comprises unsubstituted linear C1- C6 heteroalkylene.
  • L 2 comprises unsubstituted linear C1-C3 heteroalkylene.
  • L 2 comprises , wherein y is an integer between 1 and 100, inclusive.
  • y is an integer between 1 and 90, inclusive; between 1 and 80, inclusive; between 1 and 70, inclusive; between 1 and 60, inclusive; between 1 and 50, inclusive; between 1 and 40, inclusive; between 1 and 30, inclusive; between 1 and 20, inclusive; between 1 and 10, inclusive; or between 1 and 5, inclusive.
  • y is an integer between 1 and 10, inclusive.
  • y is an integer between 1 and 5, inclusive.
  • R0708.70175WO00 112/180 #13640937v1 comprises optionally substituted C 1 -C 6 heteroalkenylene.
  • L 2 comprises optionally substituted C1-C3 heteroalkenylene.
  • L 2 comprises substituted heteroalkenylene.
  • L 2 comprises substituted C1-C12 heteroalkenylene.
  • L 2 comprises substituted C 1 -C 6 heteroalkenylene.
  • L 2 comprises substituted C 1 -C 3 heteroalkenylene.
  • L 2 comprises unsubstituted heteroalkenylene.
  • L 2 comprises unsubstituted C1-C12 heteroalkenylene.
  • L 2 comprises optionally substituted 3-10 membered heterocyclylene. In some embodiments, L 2 comprises optionally substituted 3-6 membered heterocyclylene. In some embodiments, L 2 comprises substituted heterocyclylene. In some embodiments, L 2 comprises substituted 3-10 membered heterocyclylene. In some embodiments, L 2 comprises substituted 3-6 membered heterocyclylene. In some embodiments, L 2 comprises unsubstituted heterocyclylene. In some embodiments, L 2 comprises unsubstituted 3-10 membered heterocyclylene. In some embodiments, L 2 comprises unsubstituted 3-6 membered heterocyclylene. [0423] In some embodiments, L 2 comprises optionally substituted arylene.
  • L 2 comprises optionally substituted phenylene. In some embodiments, L 2 comprises substituted arylene. In some embodiments, L 2 comprises substituted phenylene. In some embodiments, L 2 comprises unsubstituted arylene. In some embodiments, L 2 comprises unsubstituted phenylene. [0424] In some embodiments, L 2 comprises optionally substituted heteroarylene. In some embodiments,
  • L 2 comprises optionally substituted 5-10 membered heteroarylene. In some embodiments, L 2 comprises optionally substituted 5-6 membered monocyclic heteroarylene. In some embodiments, L 2 comprises substituted heteroarylene. In some embodiments, L 2 comprises substituted 5-10 membered heteroarylene. In some embodiments, L 2 comprises substituted 5-6 membered monocyclic heteroarylene. In some embodiments, L 2 comprises unsubstituted heteroarylene. In some embodiments, L 2 comprises unsubstituted 5-10 membered heteroarylene. In some embodiments, L 2 comprises unsubstituted 5-6 membered monocyclic heteroarylene.
  • L 2 comprises , [0426]
  • the compound of Formula (VII) is of Formula (VII-A): or a salt thereof, wherein y is an integer between 1 and 100, inclusive, such that the sulfur atom of the cysteine residue in the azide-functionalized peptide is bonded to a group of the formula: .
  • y is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.
  • y is 1, 2, 3, 4, or 5.
  • y is 4.
  • the method further comprises contacting the azide-functionalized peptide with a compound comprising a complementary moiety, such that the azide moiety of the azide- functionalized peptide reacts with the complementary moiety via click chemistry to provide a triazole- linked conjugated peptide.
  • the method comprises contacting the azide-functionalized peptide with a compound comprising a complementary moiety, wherein the compound comprising a complementary moiety is of Formula (X): or a salt thereof, to provide a triazole-linked conjugated peptide, such that a sulfur atom of a cysteine residue in the triazole-linked conjugated peptide is bonded to a group of the formula:
  • each of R 3a and R 3b is independently hydrogen, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl, or R 3a and R 3b are joined together with their intervening atoms to form optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl.
  • the sulfur atom of the cysteine residue in the triazole-linked conjugated peptide is bonded to a group of the formula: .
  • the sulfur atom of the cysteine residue in the triazole-linked conjugated peptide is bonded to a group of the formula: .
  • a method of sequencing a peptide having at least one free cysteine residue comprising: reacting the peptide with a compound of Formula (IX): or a salt thereof, to provide an azide-functionalized peptide, such that the sulfur atom of the cysteine residue in the azide-functionalized peptide is bonded to a group of the formula: performing a peptide sequencing protocol; wherein: Y is a leaving group;
  • R0708.70175WO00 116/180 #13640937v1 L 3 is optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted heteroalkylene, optionally substituted heteroalkenylene, optionally substituted heteroalkynylene, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, optionally substituted heteroarylene, or a combination thereof; and R 4 is hydrogen, optionally substituted aliphatic, or a nitrogen protecting group.
  • Y is halo (e.g., –F, –Cl, –Br, –I).
  • Y is –Cl, –Br, or –I. In some embodiments, Y is –Cl. In some embodiments, Y is –Br. In some embodiments, Y is –I. [0436] In some embodiments, the compound of Formula (IX) is of Formula (IX-A): or a salt thereof. [0437] In some embodiments, L 3 comprises optionally substituted alkylene. In some embodiments, L 3 comprises optionally substituted C 1 -C 12 alkylene. In some embodiments, L 3 comprises optionally substituted C 1 -C 6 alkylene. In some embodiments, L 3 comprises optionally substituted C 1 -C 3 alkylene.
  • L 3 comprises substituted linear C1-C12 alkylene. In some embodiments, L 3 comprises substituted linear C1-C6 alkylene. In some embodiments, L 3 comprises substituted linear C1-C3 alkylene. In some embodiments, L 3 comprises unsubstituted alkylene. In some embodiments, L 3 comprises unsubstituted C1-C12 alkylene. In some embodiments, L 3 comprises unsubstituted C1-C6 alkylene. In some embodiments, L 3 comprises unsubstituted C1-C3 alkylene. In some embodiments, L 3 comprises unsubstituted linear alkylene. In some embodiments, L 3 comprises unsubstituted linear C1-C12 alkylene.
  • L 3 comprises substituted C1-C6 heteroalkenylene. In some embodiments, L 3 comprises substituted C1-C3 heteroalkenylene. In some embodiments, L 3 comprises unsubstituted heteroalkenylene. In some embodiments, L 3 comprises unsubstituted C1-C12 heteroalkenylene. In some embodiments, L 3 comprises unsubstituted C1-C6 heteroalkenylene. In some embodiments, L 3 comprises unsubstituted C1-C3 heteroalkenylene. [0449] In some embodiments, L 3 comprises optionally substituted heteroalkynylene. In some embodiments, L 3 comprises optionally substituted C1-C12 heteroalkynylene.
  • L 3 comprises optionally substituted C1-C6 heteroalkynylene. In some embodiments, L 3 comprises optionally substituted C1-C3 heteroalkynylene. In some embodiments, L 3 comprises substituted heteroalkynylene. In some embodiments, L 3 comprises substituted C1-C12 heteroalkynylene. In some embodiments, L 3 comprises substituted C1-C6 heteroalkynylene. In some embodiments, L 3 comprises substituted C1-C3
  • L 3 comprises unsubstituted heteroalkynylene. In some embodiments, L 3 comprises unsubstituted C1-C12 heteroalkynylene. In some embodiments, L 3 comprises unsubstituted C1-C6 heteroalkynylene. In some embodiments, L 3 comprises unsubstituted C1-C3 heteroalkynylene. [0450] In some embodiments, L 3 comprises optionally substituted carbocyclylene. In some embodiments, L 3 comprises optionally substituted C3-C10 carbocyclylene.
  • L 3 comprises optionally substituted C3-C6 carbocyclylene. In some embodiments, L 3 comprises substituted carbocyclylene. In some embodiments, L 3 comprises substituted C3-C10 carbocyclylene. In some embodiments, L 3 comprises substituted C3-C6 carbocyclylene. In some embodiments, L 3 comprises unsubstituted carbocyclylene. In some embodiments, L 3 comprises unsubstituted C3-C10 carbocyclylene. In some embodiments, L 3 comprises unsubstituted C3-C6 carbocyclylene. [0451] In some embodiments, L 3 comprises optionally substituted heterocyclylene.
  • L 3 comprises optionally substituted phenylene. In some embodiments, L 3 comprises substituted arylene. In some embodiments, L 3 comprises substituted phenylene. In some embodiments, L 3 comprises unsubstituted arylene. In some embodiments, L 3 comprises unsubstituted phenylene. [0453] In some embodiments, L 3 comprises optionally substituted heteroarylene. In some embodiments, L 3 comprises optionally substituted 5-10 membered heteroarylene. In some embodiments, L 3 comprises optionally substituted 5-6 membered monocyclic heteroarylene. In some embodiments, L 3 comprises substituted heteroarylene. In some embodiments, L 3 comprises substituted 5-10 membered heteroarylene.
  • L 3 comprises substituted 5-6 membered monocyclic heteroarylene. In some embodiments, L 3 comprises unsubstituted heteroarylene. In some embodiments, L 3 comprises unsubstituted 5-10 membered heteroarylene. In some embodiments, L 3 comprises unsubstituted 5-6 membered monocyclic heteroarylene.
  • the compound of Formula (IX) is of Formula (IX-B): or a salt thereof, wherein z is an integer between 1 and 100, inclusive, such that a sulfur atom of a cysteine residue in the azide-functionalized peptide is bonded to a group of the formula:
  • z is 3.
  • R 4 is hydrogen.
  • R 4 is optionally substituted aliphatic.
  • R 4 is optionally substituted alkyl.
  • R 4 is optionally substituted C 1-12 alkyl.
  • R 4 is a nitrogen protecting group.
  • the compound of Formula (IX) is of Formula (IX-D): or a salt thereof, wherein z is an integer between 1 and 100, inclusive, such that a sulfur atom of a cysteine residue in the azide-functionalized peptide is bonded to a group of the formula: .
  • z is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments of Formula (IX-D), z is 1, 2, 3, 4, or 5. In some embodiments of Formula (IX-D), z is 3. [0458] In some embodiments, the compound of Formula (IX) is of formula: , or a salt thereof, such that a sulfur atom of a cysteine residue in the azide-functionalized peptide is bonded to a group of the formula: . [0459] In some embodiments, the method further comprises contacting the azide-functionalized peptide with a compound comprising a complementary moiety, such that the azide moiety of the azide-
  • the method further comprises contacting the azide-functionalized peptide with a compound comprising a complementary moiety, wherein the compound comprising a complementary moiety is of Formula (X): or a salt thereof, to provide a triazole-linked conjugated peptide, such that a sulfur atom of a cysteine residue in the triazole-linked conjugated peptide is bonded to a group of the formula: , wherein: each of R 3a and R 3b is independently hydrogen, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl, or R 3a and R 3b are joined together with their intervening atoms to form optionally
  • the sulfur atom of the cysteine residue in the triazole-linked conjugated peptide is bonded to a group of the formula: .
  • the sulfur atom of the cysteine residue in the triazole-linked conjugated peptide is bonded to a group of the formula: .
  • L 2 is optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted heteroalkylene, optionally substituted heteroalkenylene, optionally substituted heteroalkynylene, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, optionally substituted heteroarylene, or a combination thereof;
  • L 3 is optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted heteroalkylene, optionally substituted heteroalkenylene, optionally substituted heteroalkynylene, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, optionally substituted heteroarylene, or a combination thereof;
  • R 4 is hydrogen, optionally substituted aliphatic, or a nitrogen protecting group.
  • R 3a and R 3b are hydrogen.
  • R 3a is hydrogen.
  • R 3b is hydrogen.
  • at least one of R 3a and R 3b is optionally substituted aliphatic.
  • R 3a is optionally substituted aliphatic.
  • R 3b is optionally substituted aliphatic.
  • At least one of R 3a and R 3b is substituted aliphatic. In some embodiments, R 3a is substituted aliphatic. In some embodiments, R 3b is substituted aliphatic. [0469] In some embodiments, at least one of R 3a and R 3b is optionally substituted alkyl. In some embodiments, at least one of R 3a and R 3b is optionally substituted C1-C12 alkyl. In some embodiments, at least one of R 3a and R 3b is optionally substituted C1-C6 alkyl. In some embodiments, at least one of R 3a and R 3b is optionally substituted C1-C3 alkyl.
  • At least one of R 3a and R 3b is optionally substituted linear alkyl. In some embodiments, at least one of R 3a and R 3b is substituted alkyl. In some embodiments, at least one of R 3a and R 3b is substituted C1-C12 alkyl. In some embodiments, at least one of R 3a and R 3b is substituted C1-C6 alkyl. In some embodiments, at least one of R 3a and R 3b is substituted C1-C3 alkyl. In some embodiments, at least one of R 3a and R 3b is substituted linear alkyl. In some embodiments, R 3a is optionally substituted alkyl. In some embodiments, R 3b is optionally substituted alkyl.
  • At least one of R 3a and R 3b is optionally substituted alkenyl. In some embodiments, at least one of R 3a and R 3b is optionally substituted C 1 -C 12 alkenyl. In some embodiments,
  • R0708.70175WO00 125/180 #13640937v1 at least one of R 3a and R 3b is optionally substituted C 1 -C 6 alkenyl. In some embodiments, at least one of R 3a and R 3b is optionally substituted C1-C3 alkenyl. In some embodiments, at least one of R 3a and R 3b is substituted alkenyl. In some embodiments, at least one of R 3a and R 3b is substituted C1-C12 alkenyl. In some embodiments, at least one of R 3a and R 3b is substituted C 1 -C 6 alkenyl. In some embodiments, at least one of R 3a and R 3b is substituted C1-C3 alkenyl.
  • R 3a is optionally substituted alkenyl. In some embodiments, R 3b is optionally substituted alkenyl. [0471] In some embodiments, at least one of R 3a and R 3b is optionally substituted alkynyl. In some embodiments, at least one of R 3a and R 3b is optionally substituted C1-C12 alkynyl. In some embodiments, at least one of R 3a and R 3b is optionally substituted C1-C6 alkynyl. In some embodiments, at least one of R 3a and R 3b is optionally substituted C1-C3 alkynyl. In some embodiments, at least one of R 3a and R 3b is substituted alkynyl.
  • At least one of R 3a and R 3b is substituted C1-C12 alkynyl. In some embodiments, at least one of R 3a and R 3b is substituted C1-C6 alkynyl. In some embodiments, at least one of R 3a and R 3b is substituted C1-C3 alkynyl. In some embodiments, R 3a is optionally substituted alkynyl. In some embodiments, R 3b is optionally substituted alkynyl. [0472] In some embodiments, at least one of R 3a and R 3b is optionally substituted heteroaliphatic. In some embodiments, R 3a is optionally substituted heteroaliphatic.
  • R 3b is optionally substituted heteroaliphatic. In some embodiments, at least one of R 3a and R 3b is substituted heteroaliphatic. In some embodiments, R 3a is substituted heteroaliphatic. In some embodiments, R 3b is substituted heteroaliphatic. [0473] In some embodiments, at least one of R 3a and R 3b is optionally substituted heteroalkyl. In some embodiments, at least one of R 3a and R 3b is optionally substituted C1-C12 heteroalkyl. In some embodiments, at least one of R 3a and R 3b is optionally substituted C1-C6 heteroalkyl.
  • At least one of R 3a and R 3b is optionally substituted C1-C3 heteroalkyl. In some embodiments, at least one of R 3a and R 3b is optionally substituted linear heteroalkyl. In some embodiments, at least one of R 3a and R 3b is substituted heteroalkyl. In some embodiments, at least one of R 3a and R 3b is substituted C 1 -C 12 heteroalkyl. In some embodiments, at least one of R 3a and R 3b is substituted C 1 -C 6 heteroalkyl. In some embodiments, at least one of R 3a and R 3b is substituted C 1 -C 3 heteroalkyl.
  • At least one of R 3a and R 3b is substituted linear heteroalkyl. In some embodiments, R 3a is optionally substituted heteroalkyl. In some embodiments, R 3b is optionally substituted heteroalkyl. [0474] In some embodiments, at least one of R 3a and R 3b is optionally substituted heteroalkenyl. In some embodiments, at least one of R 3a and R 3b is optionally substituted C 1 -C 12 heteroalkenyl. In some embodiments, at least one of R 3a and R 3b is optionally substituted C 1 -C 6 heteroalkenyl. In some embodiments, at least one of R 3a and R 3b is optionally substituted C 1 -C 3 heteroalkenyl.
  • At least one of R 3a and R 3b is substituted C1-C12 heteroalkynyl. In some embodiments, at least one of R 3a and R 3b is substituted C1-C6 heteroalkynyl. In some embodiments, at least one of R 3a and R 3b is substituted C1-C3 heteroalkynyl. In some embodiments, R 3a is optionally substituted heteroalkynyl. In some embodiments, R 3b is optionally substituted heteroalkynyl. [0476] In some embodiments, at least one of R 3a and R 3b is optionally substituted carbocyclyl.
  • At least one of R 3a and R 3b is optionally substituted C3-C10 carbocyclyl. In some embodiments, at least one of R 3a and R 3b is optionally substituted C3-C6 carbocyclyl. In some embodiments, at least one of R 3a and R 3b is substituted carbocyclyl. In some embodiments, at least one of R 3a and R 3b is substituted C3-C10 carbocyclyl. In some embodiments, at least one of R 3a and R 3b is substituted C3-C6 carbocyclyl. In some embodiments, R 3a is optionally substituted carbocyclyl. In some embodiments, R 3b is optionally substituted carbocyclyl.
  • At least one of R 3a and R 3b is optionally substituted heterocyclyl. In some embodiments, at least one of R 3a and R 3b is optionally substituted 3-10 membered heterocyclyl. In some embodiments, at least one of R 3a and R 3b is optionally substituted 3-6 membered heterocyclyl. In some embodiments, at least one of R 3a and R 3b is substituted heterocyclyl. In some embodiments, at least one of R 3a and R 3b is substituted 3-10 membered heterocyclyl. In some embodiments, at least one of R 3a and R 3b is substituted 3-6 membered heterocyclyl. In some embodiments, R 3a is optionally substituted heterocyclyl.
  • R 3b is optionally substituted heterocyclyl.
  • at least one of R 3a and R 3b is optionally substituted aryl.
  • at least one of R 3a and R 3b is optionally substituted phenyl.
  • at least one of R 3a and R 3b is substituted aryl.
  • at least one of R 3a and R 3b is substituted phenyl.
  • R 3a is optionally substituted aryl.
  • R 3b is optionally substituted aryl.
  • at least one of R 3a and R 3b is optionally substituted heteroaryl.
  • R 3a and R 3b are joined together with their intervening atoms to form substituted carbocyclyl, substituted heterocyclyl, substituted aryl, or substituted heteroaryl. In some embodiments, R 3a and R 3b are joined together with their intervening atoms to form optionally substituted carbocyclyl or optionally substituted heterocyclyl. In some embodiments, R 3a and R 3b are joined together with their intervening atoms to form substituted carbocyclyl or substituted heterocyclyl.
  • R 3a and R 3b are joined together with their intervening atoms to form optionally substituted carbocyclyl. In some embodiments, R 3a and R 3b are joined together with their intervening atoms to form optionally substituted C3-C10 carbocyclyl. In some embodiments, R 3a and R 3b are joined together with their intervening atoms to form optionally substituted unsaturated C3-C10 carbocyclyl. In some embodiments, R 3a and R 3b are joined together with their intervening atoms to form optionally substituted cyclooctenyl. In some embodiments, R 3a and R 3b are joined together with their intervening atoms to form substituted carbocyclyl.
  • R 3C is optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, or –OR A .
  • R 3C is optionally substituted aliphatic.
  • R 3C is optionally substituted heteroaliphatic.
  • R 3C is optionally substituted carbocyclyl.
  • R 3C is optionally substituted heterocyclyl.
  • R 3C is optionally substituted aryl.
  • the compound comprising a complementary moiety comprises dibenzoazacyclooctyne (DIBAC or DBCO). In some embodiments, the compound comprising a complementary moiety comprises biarylazacyclooctynone (BARAC). In some embodiments, the compound comprising a complementary moiety comprises dibenzocyclooctyne (DIBO). In some embodiments, the compound comprising a complementary moiety comprises difluorinated cyclooctyne (DIFO). In some embodiments, the compound comprising a complementary moiety comprises bicyclononyne (BCN). In some embodiments, the compound comprising a complementary moiety comprises dimethoxyazacyclooctyne (DIMAC).
  • DIBAC dibenzoazacyclooctyne
  • DBCO dibenzoazacyclooctyne
  • the compound comprising a complementary moiety comprises biarylazacyclooctynone (BARAC).
  • the polypeptidyl group comprises between 5 and 10 amino acid residues, between 5 and 15 amino acid residues, between 5 and 20 amino acid residues, between 10 and 15 amino acid residues, between 10 and 20 amino acid residues, or between 15 and 20 amino acid residues. In some embodiments, the polypeptidyl group comprises between 5 and 15 amino acid residues. [0488] In some embodiments, the polypeptidyl group has a length of at least about 20 ⁇ , 25 ⁇ , 30 ⁇ , 35 ⁇ , 40 ⁇ , 45 ⁇ , 50 ⁇ , 55 ⁇ , 60 ⁇ , 65 ⁇ , 70 ⁇ , or 75 ⁇ .
  • the polypeptidyl group has a length in a range from 20 ⁇ to 30 ⁇ , 20 ⁇ to 35 ⁇ , 20 ⁇ to 40 ⁇ , 20 ⁇ to 45 ⁇ , 20 ⁇ to 50 ⁇ , 20 ⁇ to 55 ⁇ , 20 ⁇ to 60 ⁇ , 20 ⁇ to 65 ⁇ , 20 ⁇ to 70 ⁇ , 20 ⁇ to 75 ⁇ , 30 ⁇ to 40 ⁇ , 30 ⁇ to 45 ⁇ , 30 ⁇ to 50 ⁇ , 30 ⁇ to 55 ⁇ , 30 ⁇ to 60 ⁇ , 30 ⁇ to 65 ⁇ , 30 ⁇ to 70 ⁇ , 30 ⁇ to 75 ⁇ , 40 ⁇ to 50 ⁇ , 40 ⁇ to 55 ⁇ , 40 ⁇ to 60 ⁇ , 40 ⁇ to 65 ⁇ , 40 ⁇ to 70 ⁇ , 40 ⁇ to 75 ⁇ , 50 ⁇ to 60 ⁇ , 50 ⁇ to 65 ⁇ , 50 ⁇ to 75 ⁇ , 50 ⁇ to 60 ⁇ , 50 ⁇ to 65 ⁇ , 50 ⁇ to 75 ⁇
  • the polypeptidyl group comprises at least 1 negatively charged moiety at physiological pH. In certain embodiments, the polypeptidyl group comprises at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, or at least 15 negatively charged moieties at physiological pH.
  • the polypeptidyl group comprises between 1 and 2, 1 and 3, 1 and 4, 1 and 5, 1 and 6, 1 and 7, 1 and 8, 1 and 9, 1 and 10, 1 and 11, 1 and 12, 1 and 13, 1 and 14, 1 and 15, 2 and 3, 2 and 4, 2 and 5, 2 and 6, 2 and 7, 2 and 8, 2 and 9, 2 and 10, 2 and 11, 2 and 12, 2 and 13, 2 and 14, 2 and 15, 3 and 4, 3 and 5, 3 and 6, 3 and 7, 3 and 8, 3 and 9, 3 and 10, 3 and 11, 3 and 12, 3 and 13, 3 and 14, 3 and 15, 4 and 5, 4 and 6, 4 and 7, 4 and 8, 4 and 9, 4 and 10, 4 and 11, 4 and 12, 4 and 13, 4 and 14, 4 and 15, 5 and 6, 5 and 7, 5 and 8, 5 and 9, 5 and 10, 5 and 11, 5 and 12, 5 and 13, 5 and 14, 5 and 15, 6 and 10, 6 and 15, 7 and 10, 7 and 15, 8 and 10, 8 and 15, 9 and 10, 9 and 15, or 10 and 15 negatively charged moieties at physiological pH.
  • the polypeptidyl group comprises between 1 and 10 negatively charged moieties at physiological pH. [0490] In some embodiments, the polypeptidyl group comprises at least 1 aspartate residue. In certain embodiments, the polypeptidyl group comprises at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, or at least 15 aspartate residues.
  • the polypeptidyl group comprises between 1 and 2, 1 and 3, 1 and 4, 1 and 5, 1 and 6, 1 and 7, 1 and 8, 1 and 9, 1 and 10, 1 and 11, 1 and 12, 1 and 13, 1 and 14, 1 and 15, 2 and 3, 2 and 4, 2 and 5, 2 and 6, 2 and 7, 2 and 8, 2 and 9, 2 and 10, 2 and 11, 2 and 12, 2 and 13, 2 and 14, 2 and 15, 3 and 4, 3 and 5, 3 and 6, 3 and 7, 3 and 8, 3 and 9, 3 and 10, 3 and 11, 3 and 12, 3 and 13, 3 and 14, 3 and 15, 4 and 5, 4 and 6, 4 and 7, 4 and 8, 4 and 9, 4 and 10, 4 and 11, 4 and 12, 4 and
  • the polypeptidyl group comprises between 1 and 10 aspartate residues. [0491] In some embodiments, the polypeptidyl group comprises at least 1 phenylalanine residue.
  • the polypeptidyl group comprises at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, or at least 15 phenylalanine residues.
  • the polypeptidyl group comprises between 1 and 2, 1 and 3, 1 and 4, 1 and 5, 1 and 6, 1 and 7, 1 and 8, 1 and 9, 1 and 10, 1 and 11, 1 and 12, 1 and 13, 1 and 14, 1 and 15, 2 and 3, 2 and 4, 2 and 5, 2 and 6, 2 and 7, 2 and 8, 2 and 9, 2 and 10, 2 and 11, 2 and 12, 2 and 13, 2 and 14, 2 and 15, 3 and 4, 3 and 5, 3 and 6, 3 and 7, 3 and 8, 3 and 9, 3 and 10, 3 and 11, 3 and 12, 3 and 13, 3 and 14, 3 and 15, 4 and 5, 4 and 6, 4 and 7, 4 and 8, 4 and 9, 4 and 10, 4 and 11, 4 and 12, 4 and 13, 4 and 14, 4 and 15, 5 and 6, 5 and 7, 5 and 8, 5 and 9, 5 and 10, 5 and 11, 5 and 12, 5 and 13, 5 and 14, 5 and 15, 6 and 10, 6 and 15, 7 and 10, 7 and 15, 8 and 10, 8 and 15, 9 and 10, 9 and 15, or 10 and 15 glycine residues.
  • the polypeptidyl group comprises at least 1 proline residue. In certain embodiments, the polypeptidyl group comprises at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, or at least 15 proline residues.
  • the polypeptidyl group comprises between 1 and 2, 1 and 3, 1 and 4, 1 and 5, 1 and 6, 1 and 7, 1 and 8, 1 and 9, 1 and 10, 1 and 11, 1 and 12, 1 and 13, 1 and 14, 1 and 15, 2 and 3, 2 and 4, 2 and 5, 2 and 6, 2 and 7, 2 and 8, 2 and 9, 2 and 10, 2 and 11, 2 and 12, 2 and 13, 2 and 14, 2 and 15, 3 and 4, 3 and 5, 3 and 6, 3 and 7, 3 and 8, 3 and 9, 3 and 10, 3 and 11, 3 and 12, 3 and 13, 3 and 14, 3 and 15, 4 and 5, 4 and 6, 4 and 7, 4 and 8, 4 and 9, 4 and 10, 4 and 11, 4 and 12, 4 and 13, 4 and 14, 4 and 15, 5 and 6, 5 and 7, 5 and 8, 5 and 9, 5 and 10, 5 and 11, 5 and 12, 5 and 13, 5 and 14, 5 and 15, 6 and 10, 6 and 15, 7 and 10, 7 and 15, 8 and 10, 8 and 15, 9 and 10, 9 and 15, or 10 and 15 proline residues.
  • the polypeptidyl group comprises at least 1 DD repeat, GG repeat, FF
  • the polypeptidyl group comprises between 1 and 2, 1 and 3, 1 and 4, 1 and 5, 1 and 6, 1 and 7, 1 and 8, 1 and 9, 1 and 10, 1 and 11, 1 and 12, 1 and 13, 1 and 14, 1 and 15, 2 and 3, 2 and 4, 2 and 5, 2 and 6, 2 and 7, 2 and 8, 2 and 9, 2 and 10, 2 and 11, 2 and 12, 2 and 13, 2 and 14, 2 and 15, 3 and 4, 3 and 5, 3 and 6, 3 and 7, 3 and 8, 3 and 9, 3 and 10, 3 and 11, 3 and 12, 3 and 13, 3 and 14, 3 and 15, 4 and 5, 4 and 6, 4 and 7, 4 and 8, 4 and 9, 4 and 10, 4 and 11, 4 and 12, 4 and 13, 4 and 14, 4 and 15, 5 and 6, 5 and 7, 5 and 8, 5 and 9, 5 and 10, 5 and 11, 5 and 12, 5 and 13, 5 and 14, 5 and 15, 6 and 10, 6 and 15, 7 and 10, 7 and 15, 8 and 10, 8 and 15, 9 and 10, 9 and 15, or 10 and 15 DD repeats, GG repeats, FF repeats, DDD repeats, GGG, and/or FFF repeats.
  • the polypeptidyl group comprises a sequence selected from the group consisting of GPPPPPPPPG (SEQ ID NO: 11), isoEGWRW (SEQ ID NO: 12), DDGGGDDDFF (SEQ ID NO: 13), GGSSSGSGNDEEFQ (SEQ ID NO: 14), GGGGGDPDPDFF (SEQ ID NO: 15), GDGDGDGDFF (SEQ ID NO: 16), NNGGGNNNFF (SEQ ID NO: 17), and DDGGGCyCyCyFF (SEQ ID NO: 18), or a salt thereof, wherein Cy is a cysteic acid.
  • the polypeptidyl group comprises DDGGGDDDFF (SEQ ID NO: 13).
  • R0708.70175WO00 133/180 #13640937v1 from: reacting the peptide, the azide-functionalized peptide, or the triazole-linked conjugated peptide in an enzymatic digestion process; obtaining data during the enzymatic digestion process; analyzing the data to determine portions of the data corresponding to amino acids that are sequentially exposed at a terminus of the peptide, the azide-functionalized peptide, or the triazole-linked conjugated peptide during the digestion process; and outputting an amino acid sequence representative of the peptide, the azide-functionalized peptide, or the triazole-linked conjugated peptide.
  • Maleimides are often used for cysteine conjugation, but the thiosuccinimide adduct is prone to deconjugation via retro-Michael reaction or ring-opening by hydrolysis, leading to premature drug release in plasma.
  • Tagging cysteines with arylpropiolonitrile (APN) reagents is another strategy for cysteine functionalization.
  • APN arylpropiolonitrile
  • the presence of two electron withdrawing groups may activate the substrate for reactivity with thiols. Cysteine conjugation leads to a stereoisomeric product mixture that contains mostly the Z-isomer, and the product is hydrolysis-stable compared to a thiosuccinimide addition product.
  • reaction mixture was briefly centrifuged and 5 ⁇ L of reaction mixture was diluted with 45 ⁇ L of water for TOF-LC/MS analysis.7 ⁇ L of the 50 ⁇ L mixture was injected on a LC-TOF MS Agilent 6230.
  • the azidopeptide (Tet-20 conjugated to VPN- N3) was subjected to standard SOP conjugation to the macromolecular linker for on-chip sequencing: 16- hour incubation of azidopeptide (47 ⁇ L) with 1 ⁇ L of 12.5 mM ceryltrimethylammonium bromide (CTAB; 0.25 mM final concentration) and 2 ⁇ L of 50 ⁇ M dsQ24-SSD-linker (2 ⁇ M final concentration) at 37 o C (FIGs.3, 4A-4B, 5A-5B).
  • CAB ceryltrimethylammonium bromide
  • Example 4 demonstrates that the reactivity of compounds of the present disclosure (e.g., Formulae (I′) and (I)) toward the thiol group of a C-terminal cysteine residue in a synthetic peptide enables on-chip sequencing of the synthetic peptide.
  • the prior art predominantly uses synthetic peptides with C-terminal lysine residues for such on-chip sequencing, which can cause sample heterogeneity due to difficulties in controlling chemoselectivity and regioselectivity.
  • Example 4 VPN-N3 enables sequencing of larger peptides (>3 kDa).
  • This Example describes how conjugation of compounds of the present disclosure (e.g., Formulae (I′) and (I)) to synthetic peptides greater than 3 kDa in size containing a C-terminal cysteine residue enables their on-chip sequencing.
  • Conjugation of VPN-N3 to >3 kDa protein fragments 1 ⁇ L of a 1 mM stock of rabies virus glycoprotein fragment YTIWMPENPRPGTPCDIFTNSRGKRASNG (SEQ ID NO: 3) (10 ⁇ M) was mixed with 6.6 ⁇ L of a 1.5 mM stock of VPN-N3 (10 molar equivalents) in 92.4 ⁇ L of sample buffer.
  • reaction mixture was incubated at 37 o C for 3 hours.5 ⁇ L of the reaction mixture was diluted with 45 ⁇ L of water for TOF-LC/MS analysis (FIGs.6A-6B).
  • Peptide derived from rabies virus glycoprotein (RVG) interacts specifically with the nicotinic acetylcholine receptor (AchR) on neuronal cells to enable viral entry.
  • RVG rabies virus glycoprotein
  • AchR nicotinic acetylcholine receptor
  • 1 ⁇ L of a 1 mM stock of Tau peptide fragment VQIVYKPVDLSKVTSKCGSLGNIHHKPGGGQ (SEQ ID NO: 4) (10 ⁇ M) was mixed with 6.6 ⁇ L of VPN-N 3 (10 molar equivalents) in 92.4 ⁇ L of sample buffer.
  • QP300 (ALVLIAFAQYLQQCPFEDHVK) (SEQ ID NO: 5), which corresponds to a Lys-C peptide fragment from human serum albumin (HSA) protein, was synthesized according to standard Fmoc solid phase chemistry.1.5 mg of QP300 in 620 ⁇ L of 1:1 acetonitrile/water generates a 1 mM QP300 stock.1 ⁇ L of 1 mM QP300 (10 ⁇ M) was mixed with 2 ⁇ L of 1 mM APN-N3 in 97 ⁇ L of sample buffer.
  • APN-N3 APN-N3
  • reaction mixture was incubated at 37 o C for 4 hours.5 ⁇ L of the reaction mixture was diluted with 45 ⁇ L of water for TOF-LC/MS analysis (FIGs.8A-8B).
  • 1 ⁇ L of 1 mM QP300 was also digested with Glu-C protease (1:100 final enzyme: protein substrate ratio) for 16 hours in 97 ⁇ L of sample buffer at 37 o C to generate ALVLIAFAQYLQQCPFE (SEQ ID NO: 24).2 ⁇ L of 1 mM APN-N3 was added to the digestion mixture and the reaction mixture was incubated at 37 o C for 3 hours.5 ⁇ L of the reaction mixture was diluted with 45 ⁇ L of water for TOF- LC/MS analysis (FIGs.9A-9B).
  • the mixture containing conjugated HSA was subjected to digestion with Lys-C (2 ⁇ L of 0.5 ⁇ g/ ⁇ L stock) for 16 hours at 37 o C. Then, 47 ⁇ L of the digestion mixture was mixed with 1 ⁇ L of 12.5 mM CTAB (0.25 mM final concentration) and 2 ⁇ L of 50 ⁇ M dsQ24-SSD-linker (2 ⁇ M final concentration) for on-chip sequencing.
  • QL847 Conjugation of VPN-N3 to HSA followed by digestion 10 ⁇ L of 200 ⁇ M HSA (20 ⁇ M final concentration) was mixed with 6 ⁇ L of 3.3 mM VPN-N3 (10 equivalents) in 100 ⁇ L of sample buffer.
  • the reaction proceeded for 4 hours at 37 o C.
  • the conjugated HSA mixture was subjected to digestion with Lys-C (2 ⁇ L of 0.5 ⁇ g/ ⁇ L stock) for 16 hours at 37 o C.
  • 47 ⁇ L of the digestion mixture was mixed with 1 ⁇ L of 12.5 mM CTAB (0.25 mM final concentration) and 2 ⁇ L of 50 ⁇ M dsQ24-SSD-linker (2 ⁇ M final concentration) for on-chip sequencing.
  • Example 6 further demonstrates the enrichment of a cysteine-containing protein in a sample (e.g., the enrichment of HSA within a sample).
  • Example 6 VPN-N3 enables sequencing of proteins with a free cysteine.
  • This Example describes how conjugation of compounds of the present disclosure (e.g., Formulae (I′), (I), and (VII)) to proteins containing a free cysteine residue (i.e., a cysteine residue not ordinarily present as a disulfide) or their digests enables on-chip sequencing of the proteins containing the free cysteine residue, under various conditions and types of workflow (e.g., order and type of digestion step).
  • the peptide mixture was then conjugated with 2.5 ⁇ L of 1 mM APN-N3 for 4 hours at 37 o C. Then, 47 ⁇ L of the digestion mixture was mixed with 1 ⁇ L of 12.5 mM CTAB (0.25 mM final concentration) and 2 ⁇ L of 50 ⁇ M dsQ24-SSD-linker (2 ⁇ M final concentration) for on-chip sequencing (FIGs.15B, 19, 20, 21A).

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Biochemistry (AREA)
  • Analytical Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biophysics (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Peptides Or Proteins (AREA)

Abstract

L'invention concerne des composés représentés par les formules (I'), (I), (IV') et (IV). L'invention concerne également des procédés de préparation des formules (Ill-i), (VI) et (VIII). L'invention concerne en outre des procédés de fonctionnalisation ou de séquençage d'un peptide ayant au moins un résidu cystéine libre, et des procédés d'enrichissement d'un échantillon comprenant un ou plusieurs peptides.
PCT/US2025/015581 2024-02-12 2025-02-12 Conjugaison de cystéine Pending WO2025174876A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US202463552520P 2024-02-12 2024-02-12
US63/552,520 2024-02-12

Publications (1)

Publication Number Publication Date
WO2025174876A1 true WO2025174876A1 (fr) 2025-08-21

Family

ID=96773460

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2025/015581 Pending WO2025174876A1 (fr) 2024-02-12 2025-02-12 Conjugaison de cystéine

Country Status (1)

Country Link
WO (1) WO2025174876A1 (fr)

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20170276686A1 (en) * 2014-09-15 2017-09-28 Board Of Regents, The University Of Texas System Single molecule peptide sequencing
US20180064829A1 (en) * 2015-04-02 2018-03-08 University Of Southern California Fluorescent bisphosphonate analogs
US20210221839A1 (en) * 2020-01-21 2021-07-22 Quantum-Si Incorporated Compounds and methods for selective c-terminal labeling
CN117384089A (zh) * 2023-10-09 2024-01-12 深圳技术大学 一种可用于蛋白质半胱氨酸残基可逆化学修饰的探针及其制备方法

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20170276686A1 (en) * 2014-09-15 2017-09-28 Board Of Regents, The University Of Texas System Single molecule peptide sequencing
US20180064829A1 (en) * 2015-04-02 2018-03-08 University Of Southern California Fluorescent bisphosphonate analogs
US20210221839A1 (en) * 2020-01-21 2021-07-22 Quantum-Si Incorporated Compounds and methods for selective c-terminal labeling
CN117384089A (zh) * 2023-10-09 2024-01-12 深圳技术大学 一种可用于蛋白质半胱氨酸残基可逆化学修饰的探针及其制备方法

Similar Documents

Publication Publication Date Title
US20240100177A1 (en) Antibody-oligonucleotide complexes and uses thereof
US20210188787A1 (en) Dota compounds and uses thereof
US20170183378A1 (en) Trapping reagents for reactive metabolites screening
JP7676302B2 (ja) バイコンジュゲート可能な標識および使用方法
JP2008537544A (ja) アブラインビルアミド、ステファシジンｂおよびそれらのアナログの合成
US20240294957A1 (en) Enzymatic synthesis of kavalactones and flavokavains
US11674136B2 (en) DNA-templated macrocycle library
US20230391799A1 (en) Fluorescent dye for protein or nucleic acid labelling
WO2025174876A1 (fr) Conjugaison de cystéine
US20230028318A1 (en) Fluorogenic amino acids
WO2024187020A1 (fr) Composés de liaison à une kinase bifonctionnelle et leurs utilisations
US20240262851A1 (en) Cyclopropene phosphoramidites and conjugates thereof
US20240360076A1 (en) Nonionic surfactants
US20170298402A1 (en) Self-labeling nucleic acids and methods of use
US20240228671A1 (en) Polypeptidyl linkers
US20250346943A1 (en) DEGRADER COMPOUNDS OF QSOX1 mRNA
US20250341526A1 (en) Novel dyes
KR20250169633A (ko) 폴리펩티딜 링커
WO2025006929A2 (fr) Colorants benzothiazole

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 25755568

Country of ref document: EP

Kind code of ref document: A1