A biomimetic polyketide-inspired approach to small-molecule ligand discovery

Journal name:
Nature Chemistry
Volume:
4,
Pages:
99–104
Year published:
DOI:
doi:10.1038/nchem.1200
Received
Accepted
Published online

Abstract

The discovery of new compounds for the pharmacological manipulation of protein function often embraces the screening of compound collections, and it is widely recognized that natural products offer beneficial characteristics as protein ligands. Much effort has therefore been focused on ‘natural product-like’ libraries, yet the synthesis and screening of such libraries is often limited by one or more of the following: modest library sizes and structural diversity, conformational heterogeneity and the costs associated with the substantial infrastructure of modern high-throughput screening centres. Here, we describe the design and execution of an approach to this broad problem by merging principles associated with biologically inspired oligomerization and the structure of polyketide-derived natural products. A novel class of chiral and conformationally constrained oligomers is described (termed ‘chiral oligomers of pentenoic amides’, COPA), which offers compatibility with split-and-pool methods and can be screened en masse in a batch mode. We demonstrate that a COPA library containing 160,000 compounds is a useful source of novel protein ligands by identifying a non-covalent synthetic ligand to the DNA-binding domain of the p53 transcription factor.

At a glance

Figures

  1. Natural and synthetic oligomers, polyketide-derived natural products and a polyketide-inspired class of chiral and conformationally rigid synthetic oligomers.
    Figure 1: Natural and synthetic oligomers, polyketide-derived natural products and a polyketide-inspired class of chiral and conformationally rigid synthetic oligomers.

    a, Selection of biological and biopolymer mimetics. b, Selection of polyketide-derived natural products (regions in red are substituted alkenes that impart conformational rigidification as a result of A(1,3) strain). c, General structure of COPAs (chiral oligomers of N-substituted 5-amino-2,4-dialkyl-3-pentenoic amides). d, Structural features that lead to the rigidification of COPA oligomers.

  2. Stereochemistry of the COPA backbone is anticipated to have a substantial impact on skeletal shape and the disposition of side chains in space.
    Figure 2: Stereochemistry of the COPA backbone is anticipated to have a substantial impact on skeletal shape and the disposition of side chains in space.

    Distribution of conformers found within 1.1 kcal mol−1 of the lowest-energy conformation identified. Calculations were conducted with Spartan-08/MMFF model/Conformer Distribution option/Monte Carlo algorithm. Coloured spheres highlight the relative position of heteroatoms (green) and alkenes (blue). Although these molecular mechanics calculations are not interpreted to predict the solution-phase structure of these oligomers, the calculations provide a uniform lens through which to observe unique characteristics associated with this new class of synthetic oligomer.

  3. Chemical development of COPA oligomers from a general oligomerization strategy, asymmetric synthesis and library construction.
    Figure 3: Chemical development of COPA oligomers from a general oligomerization strategy, asymmetric synthesis and library construction.

    a, ‘Sub-monomer’ style synthesis of peptoids. b, Asymmetric synthesis of 5-chloro-2,4-dimethyl-3-pentenoic acid 1. c, Use of 1 in solution-phase oligomerization. d, Panel of monomers used in library synthesis. e, General information regarding resin and linker used in solid-phase library synthesis. f, General structure of libraries prepared from building blocks depicted in d (COPA and peptoid tetramers).

  4. COPA library synthesis, screening, structure elucidation and validation.
    Figure 4: COPA library synthesis, screening, structure elucidation and validation.

    a, General scheme for on-bead screening of the COPA library against the DNA-binding domain of p53 (p53–DBD, residues 94–312) expressed with an epitope tag FLAG. TentaGel beads bound to p53–DBD protein were visualized under a fluorescent microscope by treating beads with anti-FLAG primary antibody and anti-IgG secondary antibody conjugated to quantum dots emitting red fluorescent light at 655 nm. b, Sequence elucidation and identification of a COPA tetramer that binds to the p53–DBD. The sequence of the COPA tetramer was established by analysis of mass spectral data derived from ETD-based fragmentation. c, Fluorescence polarization assay for the binding affinity of the fluorescein-conjugated COPA tetramer (14a) against p53–DBD, carbonic anhydrolase II (CAH II from bovine erythrocyte), platelet activating factor acetyl hydrolase (PAFAHIB3) and bromodomain containing four (BRD4) proteins. Error bars indicate standard deviation of data obtained from three independent experiments. Data analysis and curve fitting were performed with GraphPad Prism 5.0 (GraphPad Software) using a nonlinear regression method (y = [x/(Kd + x)](ymax − ymin) + ymin). A COPA tetramer with the same linker region and different side chains on the amide nitrogens was used as a control oligomer (co). The binding affinity of COPA tetramer to p53–DBD was determined as Kd ≈ 10 µM. See Supplementary Information for additional details.

Compounds

59 compounds View all compounds
  1. (R,E)-5-Chloro-2,4-dimethylpent-3-enoic acid
    Compound (R)-1 (R,E)-5-Chloro-2,4-dimethylpent-3-enoic acid
  2. (S,E)-5-Chloro-2,4-dimethylpent-3-enoic acid
    Compound (S)-1 (S,E)-5-Chloro-2,4-dimethylpent-3-enoic acid
  3. (S,E)-5-Chloro-2-(CD3)-methyl-4-methylpent-3-enoic acid
    Compound (S)-1-D3 (S,E)-5-Chloro-2-(CD3)-methyl-4-methylpent-3-enoic acid
  4. (R)-(-)-4-Benzyl-2-oxazolidinone
    Compound (R)-4 (R)-(-)-4-Benzyl-2-oxazolidinone
  5. (S)-(+)-4-Benzyl-2-oxazolidinone
    Compound (S)-4 (S)-(+)-4-Benzyl-2-oxazolidinone
  6. (S)-4-Benzyl-3-propionyl-2-oxazolidinone
    Compound (S)-5 (S)-4-Benzyl-3-propionyl-2-oxazolidinone
  7. (R)-4-Benzyl-3-propionyl-2-oxazolidinone
    Compound (R)-5 (R)-4-Benzyl-3-propionyl-2-oxazolidinone
  8. (R)-4-Benzyl-3-(CD3)-propionyl-2-oxazolidinone
    Compound (R)-5-D3 (R)-4-Benzyl-3-(CD3)-propionyl-2-oxazolidinone
  9. Methacrolein
    Compound 6 Methacrolein
  10. (4R)-4-Benzyl-3-[(2S,3R)-2,4-dimethyl-3-(trimethylsiloxy)pent-4-enoyl)]-oxazolidin-2-one
    Compound (R)-7 (4R)-4-Benzyl-3-[(2S,3R)-2,4-dimethyl-3-(trimethylsiloxy)pent-4-enoyl)]-oxazolidin-2-one
  11. (4R)-4-Benzyl-3-[(2S,3R)-2-(CD3)-methyl-4-methyl-3-(trimethylsiloxy)pent-4-enoyl)]-oxazolidin-2-one
    Compound (R)-7-D3 (4R)-4-Benzyl-3-[(2S,3R)-2-(CD3)-methyl-4-methyl-3-(trimethylsiloxy)pent-4-enoyl)]-oxazolidin-2-one
  12. (4S)-4-Benzyl-3-[(2R,3S)-2,4-dimethyl-3-(trimethylsiloxy) pent-4-enoyl)]-oxazolidin-2-one
    Compound (S)-7 (4S)-4-Benzyl-3-[(2R,3S)-2,4-dimethyl-3-(trimethylsiloxy) pent-4-enoyl)]-oxazolidin-2-one
  13. (4R)-4-Benzyl-3- [(R,S)-5-chloro-2,4-dimethylpent-3-enoyl] oxazolidin-2-one
    Compound (R)-8 (4R)-4-Benzyl-3- [(R,S)-5-chloro-2,4-dimethylpent-3-enoyl] oxazolidin-2-one
  14. (4R)-4-Benzyl-3-[(R,S)-5-chloro-2-(CD3)-methyl-4-methylpent-3-enoyl] oxazolidin-2-one
    Compound (R)-8-D3 (4R)-4-Benzyl-3-[(R,S)-5-chloro-2-(CD3)-methyl-4-methylpent-3-enoyl] oxazolidin-2-one
  15. (4S)-4-Benzyl-3-[(R,E)-5-chloro-2,4-dimethylpent-3-enoyl]oxazolidin-2-one
    Compound (S)-8 (4S)-4-Benzyl-3-[(R,E)-5-chloro-2,4-dimethylpent-3-enoyl]oxazolidin-2-one
  16. Pyrrolidine
    Compound 9 Pyrrolidine
  17. 1-(R,E)-5-Chloro-2,4-dimethylpent-3-enoyl-pyrrolidine
    Compound 10 1-(R,E)-5-Chloro-2,4-dimethylpent-3-enoyl-pyrrolidine
  18. 1-(R,E)-5-N-Benzylamino-2,4-dimethylpent-3-enoyl-pyrrolidine
    Compound 11 1-(R,E)-5-N-Benzylamino-2,4-dimethylpent-3-enoyl-pyrrolidine
  19. (R,E)-N-Benzyl-N-[(R,E)-2,4-dimethyl-5-oxo-5-(pyrrolidin-1-yl) pent-3-en-5-yl]-5-chloro-2,4-dimethylpent-3-enamide
    Compound 12 (R,E)-N-Benzyl-N-[(R,E)-2,4-dimethyl-5-oxo-5-(pyrrolidin-1-yl) pent-3-en-5-yl]-5-chloro-2,4-dimethylpent-3-enamide
  20. (S,E)-N-Benzyl-N-[(R,E)-2,4-dimethyl-5-oxo-5-(pyrrolidin-1-yl) pent-3-en-5-yl] -5-chloro-2,4-dimethylpent-3-enamide
    Compound 13 (S,E)-N-Benzyl-N-[(R,E)-2,4-dimethyl-5-oxo-5-(pyrrolidin-1-yl) pent-3-en-5-yl] -5-chloro-2,4-dimethylpent-3-enamide
  21. (2S,E)-N-((16S,17E,22S,23E)-1-Amino-20-benzyl-8-(4-bromophenethyl)-14-(3-isopropoxypropyl)-11-(3-methoxypropyl)-16,18,22,24-tetramethyl-6,9,12,15,21-pentaoxo-5-(2-oxo-2-((2-oxotetrahydrofuran-3-yl)amino)ethyl)-5,8,11,14,20-pentaazapentacosa-17,23-dien-25-yl)-N-(benzo[d][1,3]dioxol-5-ylmethyl)-5-((R,E)-N-(benzo[d][1,3]dioxol-5-ylmethyl)-5-((benzo[d][1,3]dioxol-5-ylmethyl)amino)-2,4-dimethylpent-3-enamido)-2,4-dimethylpent-3-enamide
    Compound 14 (2S,E)-N-((16S,17E,22S,23E)-1-Amino-20-benzyl-8-(4-bromophenethyl)-14-(3-isopropoxypropyl)-11-(3-methoxypropyl)-16,18,22,24-tetramethyl-6,9,12,15,21-pentaoxo-5-(2-oxo-2-((2-oxotetrahydrofuran-3-yl)amino)ethyl)-5,8,11,14,20-pentaazapentacosa-17,23-dien-25-yl)-N-(benzo[d][1,3]dioxol-5-ylmethyl)-5-((R,E)-N-(benzo[d][1,3]dioxol-5-ylmethyl)-5-((benzo[d][1,3]dioxol-5-ylmethyl)amino)-2,4-dimethylpent-3-enamido)-2,4-dimethylpent-3-enamide
  22. 5-(3-(((4E,6R,10E,12S,16E,18S,22E,24S,32R)-1-(Benzo[d][1,3]dioxol-5-yl)-8,14-bis(benzo[d][1,3]dioxol-5-ylmethyl)-20-benzyl-32-carbamoyl-4,6,10,12,16,18,22,24-octamethyl-7,13,19,25,30-pentaoxo-2,8,14,20,26,31-hexaazatritriaconta-4,10,16,22-tetraen-33-yl)thio)-2,5-dioxopyrrolidin-1-yl)-2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)benzoic acid
    Compound 14a 5-(3-(((4E,6R,10E,12S,16E,18S,22E,24S,32R)-1-(Benzo[d][1,3]dioxol-5-yl)-8,14-bis(benzo[d][1,3]dioxol-5-ylmethyl)-20-benzyl-32-carbamoyl-4,6,10,12,16,18,22,24-octamethyl-7,13,19,25,30-pentaoxo-2,8,14,20,26,31-hexaazatritriaconta-4,10,16,22-tetraen-33-yl)thio)-2,5-dioxopyrrolidin-1-yl)-2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)benzoic acid
  23. 5-(3-(((4E,6R,10E,12S,16E,18S,22E,24R,32R)-1-(Benzo[d][1,3]dioxol-5-yl)-8,14-bis(benzo[d][1,3]dioxol-5-ylmethyl)-20-benzyl-32-carbamoyl-4,6,10,12,16,18,22,24-octamethyl-7,13,19,25,30-pentaoxo-2,8,14,20,26,31-hexaazatritriaconta-4,10,16,22-tetraen-33-yl)thio)-2,5-dioxopyrrolidin-1-yl)-2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)benzoic acid
    Compound 14a-1 5-(3-(((4E,6R,10E,12S,16E,18S,22E,24R,32R)-1-(Benzo[d][1,3]dioxol-5-yl)-8,14-bis(benzo[d][1,3]dioxol-5-ylmethyl)-20-benzyl-32-carbamoyl-4,6,10,12,16,18,22,24-octamethyl-7,13,19,25,30-pentaoxo-2,8,14,20,26,31-hexaazatritriaconta-4,10,16,22-tetraen-33-yl)thio)-2,5-dioxopyrrolidin-1-yl)-2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)benzoic acid
  24. 5-(3-(((4E,6R,10E,12S,16E,18R,22E,24S,32R)-1-(Benzo[d][1,3]dioxol-5-yl)-8,14-bis(benzo[d][1,3]dioxol-5-ylmethyl)-20-benzyl-32-carbamoyl-4,6,10,12,16,18,22,24-octamethyl-7,13,19,25,30-pentaoxo-2,8,14,20,26,31-hexaazatritriaconta-4,10,16,22-tetraen-33-yl)thio)-2,5-dioxopyrrolidin-1-yl)-2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)benzoic acid
    Compound 14a-2 5-(3-(((4E,6R,10E,12S,16E,18R,22E,24S,32R)-1-(Benzo[d][1,3]dioxol-5-yl)-8,14-bis(benzo[d][1,3]dioxol-5-ylmethyl)-20-benzyl-32-carbamoyl-4,6,10,12,16,18,22,24-octamethyl-7,13,19,25,30-pentaoxo-2,8,14,20,26,31-hexaazatritriaconta-4,10,16,22-tetraen-33-yl)thio)-2,5-dioxopyrrolidin-1-yl)-2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)benzoic acid
  25. 5-(3-(((4E,6R,10E,12R,16E,18S,22E,24S,32R)-1-(Benzo[d][1,3]dioxol-5-yl)-8,14-bis(benzo[d][1,3]dioxol-5-ylmethyl)-20-benzyl-32-carbamoyl-4,6,10,12,16,18,22,24-octamethyl-7,13,19,25,30-pentaoxo-2,8,14,20,26,31-hexaazatritriaconta-4,10,16,22-tetraen-33-yl)thio)-2,5-dioxopyrrolidin-1-yl)-2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)benzoic acid
    Compound 14a-3 5-(3-(((4E,6R,10E,12R,16E,18S,22E,24S,32R)-1-(Benzo[d][1,3]dioxol-5-yl)-8,14-bis(benzo[d][1,3]dioxol-5-ylmethyl)-20-benzyl-32-carbamoyl-4,6,10,12,16,18,22,24-octamethyl-7,13,19,25,30-pentaoxo-2,8,14,20,26,31-hexaazatritriaconta-4,10,16,22-tetraen-33-yl)thio)-2,5-dioxopyrrolidin-1-yl)-2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)benzoic acid
  26. 5-(3-(((4E,6S,10E,12S,16E,18S,22E,24S,32R)-1-(Benzo[d][1,3]dioxol-5-yl)-8,14-bis(benzo[d][1,3]dioxol-5-ylmethyl)-20-benzyl-32-carbamoyl-4,6,10,12,16,18,22,24-octamethyl-7,13,19,25,30-pentaoxo-2,8,14,20,26,31-hexaazatritriaconta-4,10,16,22-tetraen-33-yl)thio)-2,5-dioxopyrrolidin-1-yl)-2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)benzoic acid
    Compound 14a-4 5-(3-(((4E,6S,10E,12S,16E,18S,22E,24S,32R)-1-(Benzo[d][1,3]dioxol-5-yl)-8,14-bis(benzo[d][1,3]dioxol-5-ylmethyl)-20-benzyl-32-carbamoyl-4,6,10,12,16,18,22,24-octamethyl-7,13,19,25,30-pentaoxo-2,8,14,20,26,31-hexaazatritriaconta-4,10,16,22-tetraen-33-yl)thio)-2,5-dioxopyrrolidin-1-yl)-2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)benzoic acid
  27. (R,E)-5-Chloro-2,4-dimethylpent-3-enoyl-(L)-proline methyl ester
    Compound 15 (R,E)-5-Chloro-2,4-dimethylpent-3-enoyl-(L)-proline methyl ester
  28. (S,E)-5-Chloro-2,4-dimethylpent-3-enoyl-(L)-proline methyl ester
    Compound 16 (S,E)-5-Chloro-2,4-dimethylpent-3-enoyl-(L)-proline methyl ester
  29. (6E,8R,12E,14R,18E,20R,24E,26S)-16-(2-Acetamidoethyl)-41-amino-34-(4-bromophenethyl)-10,22-diisopentyl-28-(3-isopropoxypropyl)-31-(3-methoxypropyl)-6,8,12,14,18,20,24,26-octamethyl-9,15,21,27,30,33,36-heptaoxo-37-(2-oxo-2-((2-oxotetrahydrofuran-3-yl)amino)ethyl)-4,10,16,22,28,31,34,37-octaazahentetraconta-6,12,18,24-tetraen-1-oic acid
    Compound 17 (6E,8R,12E,14R,18E,20R,24E,26S)-16-(2-Acetamidoethyl)-41-amino-34-(4-bromophenethyl)-10,22-diisopentyl-28-(3-isopropoxypropyl)-31-(3-methoxypropyl)-6,8,12,14,18,20,24,26-octamethyl-9,15,21,27,30,33,36-heptaoxo-37-(2-oxo-2-((2-oxotetrahydrofuran-3-yl)amino)ethyl)-4,10,16,22,28,31,34,37-octaazahentetraconta-6,12,18,24-tetraen-1-oic acid
  30. (2R,10S,11E,16R,17E,22R,23E,28R,29E)-20-(2-Acetamidoethyl)-1-amino-2-(((1-(3-carboxy-4-(6-hydroxy-3-oxo-3H-xanthen-9-yl)phenyl)-2,5-dioxopyrrolidin-3-yl)thio)methyl)-14,26-diisopentyl-10,12,16,18,22,24,28,30-octamethyl-1,4,9,15,21,27-hexaoxo-3,8,14,20,26,32-hexaazapentatriaconta-11,17,23,29-tetraen-35-oic acid
    Compound 17a (2R,10S,11E,16R,17E,22R,23E,28R,29E)-20-(2-Acetamidoethyl)-1-amino-2-(((1-(3-carboxy-4-(6-hydroxy-3-oxo-3H-xanthen-9-yl)phenyl)-2,5-dioxopyrrolidin-3-yl)thio)methyl)-14,26-diisopentyl-10,12,16,18,22,24,28,30-octamethyl-1,4,9,15,21,27-hexaoxo-3,8,14,20,26,32-hexaazapentatriaconta-11,17,23,29-tetraen-35-oic acid
  31. (6E,8R,12E,14R)-29-Amino-22-(4-bromophenethyl)-10-(2-carboxyethyl)-19-(3-hydroxypropyl)-16-(3-isopropoxypropyl)-4-((R,E)-5-((S,E)-N-(4-methoxybenzyl)-5-((4-methoxybenzyl)amino)-2,4-dimethylpent-3-enamido)-2,4-dimethylpent-3-enoyl)-6,8,12,14-tetramethyl-9,15,18,21,24-pentaoxo-25-(2-oxo-2-((2-oxotetrahydrofuran-3-yl)amino)ethyl)-4,10,16,19,22,25-hexaazanonacosa-6,12-dien-1-oic acid
    Compound 18 (6E,8R,12E,14R)-29-Amino-22-(4-bromophenethyl)-10-(2-carboxyethyl)-19-(3-hydroxypropyl)-16-(3-isopropoxypropyl)-4-((R,E)-5-((S,E)-N-(4-methoxybenzyl)-5-((4-methoxybenzyl)amino)-2,4-dimethylpent-3-enamido)-2,4-dimethylpent-3-enoyl)-6,8,12,14-tetramethyl-9,15,18,21,24-pentaoxo-25-(2-oxo-2-((2-oxotetrahydrofuran-3-yl)amino)ethyl)-4,10,16,19,22,25-hexaazanonacosa-6,12-dien-1-oic acid
  32. 5-(3-(((4E,6S,10E,12R,16E,18R,22E,24R,32R)-32-Carbamoyl-14,20-bis(2-carboxyethyl)-8-(4-methoxybenzyl)-1-(4-methoxyphenyl)-4,6,10,12,16,18,22,24-octamethyl-7,13,19,25,30-pentaoxo-2,8,14,20,26,31-hexaazatritriaconta-4,10,16,22-tetraen-33-yl)thio)-2,5-dioxopyrrolidin-1-yl)-2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)benzoic acid
    Compound 18a 5-(3-(((4E,6S,10E,12R,16E,18R,22E,24R,32R)-32-Carbamoyl-14,20-bis(2-carboxyethyl)-8-(4-methoxybenzyl)-1-(4-methoxyphenyl)-4,6,10,12,16,18,22,24-octamethyl-7,13,19,25,30-pentaoxo-2,8,14,20,26,31-hexaazatritriaconta-4,10,16,22-tetraen-33-yl)thio)-2,5-dioxopyrrolidin-1-yl)-2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)benzoic acid
  33. (2R,E)-N-(2-Acetamidoethyl)-N-((16R,17E,22S,23E,28R,29E)-26-(2-acetamidoethyl)-1-amino-20-benzyl-8-(4-bromophenethyl)-14-(3-isopropoxypropyl)-11-(3-methoxypropyl)-16,18,22,24,28,30-hexamethyl-6,9,12,15,21,27-hexaoxo-5-(2-oxo-2-((2-oxotetrahydrofuran-3-yl)amino)ethyl)-5,8,11,14,20,26-hexaazahentriaconta-17,23,29-trien-31-yl)-5-((2-acetamidoethyl)amino)-2,4-dimethylpent-3-enamide
    Compound 19 (2R,E)-N-(2-Acetamidoethyl)-N-((16R,17E,22S,23E,28R,29E)-26-(2-acetamidoethyl)-1-amino-20-benzyl-8-(4-bromophenethyl)-14-(3-isopropoxypropyl)-11-(3-methoxypropyl)-16,18,22,24,28,30-hexamethyl-6,9,12,15,21,27-hexaoxo-5-(2-oxo-2-((2-oxotetrahydrofuran-3-yl)amino)ethyl)-5,8,11,14,20,26-hexaazahentriaconta-17,23,29-trien-31-yl)-5-((2-acetamidoethyl)amino)-2,4-dimethylpent-3-enamide
  34. 5-(3-(((8E,10R,14E,16R,20E,22S,26E,28R,36R)-12,18-Bis(2-acetamidoethyl)-24-benzyl-36-carbamoyl-8,10,14,16,20,22,26,28-octamethyl-2,11,17,23,29,34-hexaoxo-3,6,12,18,24,30,35-heptaazaheptatriaconta-8,14,20,26-tetraen-37-yl)thio)-2,5-dioxopyrrolidin-1-yl)-2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)benzoic acid
    Compound 19a 5-(3-(((8E,10R,14E,16R,20E,22S,26E,28R,36R)-12,18-Bis(2-acetamidoethyl)-24-benzyl-36-carbamoyl-8,10,14,16,20,22,26,28-octamethyl-2,11,17,23,29,34-hexaoxo-3,6,12,18,24,30,35-heptaazaheptatriaconta-8,14,20,26-tetraen-37-yl)thio)-2,5-dioxopyrrolidin-1-yl)-2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)benzoic acid
  35. (6E,8R,12E,14R,18E,20S,24E,26R)-22-(3-(1H-Imidazol-1-yl)propyl)-41-amino-34-(4-bromophenethyl)-28-(3-isopropoxypropyl)-31-(3-methoxypropyl)-6,8,12,14,18,20,24,26-octamethyl-9,15,21,27,30,33,36-heptaoxo-37-(2-oxo-2-((2-oxotetrahydrofuran-3-yl)amino)ethyl)-16-(3-(2-oxopyrrolidin-1-yl)propyl)-10-(3-(pyrrolidin-1-yl)propyl)-4,10,16,22,28,31,34,37-octaazahentetraconta-6,12,18,24-tetraen-1-oic acid
    Compound 20 (6E,8R,12E,14R,18E,20S,24E,26R)-22-(3-(1H-Imidazol-1-yl)propyl)-41-amino-34-(4-bromophenethyl)-28-(3-isopropoxypropyl)-31-(3-methoxypropyl)-6,8,12,14,18,20,24,26-octamethyl-9,15,21,27,30,33,36-heptaoxo-37-(2-oxo-2-((2-oxotetrahydrofuran-3-yl)amino)ethyl)-16-(3-(2-oxopyrrolidin-1-yl)propyl)-10-(3-(pyrrolidin-1-yl)propyl)-4,10,16,22,28,31,34,37-octaazahentetraconta-6,12,18,24-tetraen-1-oic acid
  36. (2R,10R,11E,16S,17E,22R,23E,28S,29E)-14-(3-(1H-Imidazol-1-yl)propyl)-1-amino-2-(((1-(3-carboxy-4-(6-hydroxy-3-oxo-3H-xanthen-9-yl)phenyl)-2,5-dioxopyrrolidin-3-yl)thio)methyl)-10,12,16,18,22,24,28,30-octamethyl-1,4,9,15,21,27-hexaoxo-20,26-bis(3-(2-oxopyrrolidin-1-yl)propyl)-3,8,14,20,26,32-hexaazapentatriaconta-11,17,23,29-tetraen-35-oic acid
    Compound 20a (2R,10R,11E,16S,17E,22R,23E,28S,29E)-14-(3-(1H-Imidazol-1-yl)propyl)-1-amino-2-(((1-(3-carboxy-4-(6-hydroxy-3-oxo-3H-xanthen-9-yl)phenyl)-2,5-dioxopyrrolidin-3-yl)thio)methyl)-10,12,16,18,22,24,28,30-octamethyl-1,4,9,15,21,27-hexaoxo-20,26-bis(3-(2-oxopyrrolidin-1-yl)propyl)-3,8,14,20,26,32-hexaazapentatriaconta-11,17,23,29-tetraen-35-oic acid
  37. (6E,8S,12E,14R)-4-((R,E)-5-((R,E)-N-(2-Acetamidoethyl)-2,4-dimethyl-5-((3-(2-oxopyrrolidin-1-yl)propyl)amino)pent-3-enamido)-2,4-dimethylpent-3-enoyl)-29-amino-10-(3-aminopropyl)-22-(4-bromophenethyl)-16-(3-isopropoxypropyl)-19-(3-methoxypropyl)-6,8,12,14-tetramethyl-9,15,18,21,24-pentaoxo-25-(2-oxo-2-((2-oxotetrahydrofuran-3-yl)amino)ethyl)-4,10,16,19,22,25-hexaazanonacosa-6,12-dien-1-oic acid
    Compound 21 (6E,8S,12E,14R)-4-((R,E)-5-((R,E)-N-(2-Acetamidoethyl)-2,4-dimethyl-5-((3-(2-oxopyrrolidin-1-yl)propyl)amino)pent-3-enamido)-2,4-dimethylpent-3-enoyl)-29-amino-10-(3-aminopropyl)-22-(4-bromophenethyl)-16-(3-isopropoxypropyl)-19-(3-methoxypropyl)-6,8,12,14-tetramethyl-9,15,18,21,24-pentaoxo-25-(2-oxo-2-((2-oxotetrahydrofuran-3-yl)amino)ethyl)-4,10,16,19,22,25-hexaazanonacosa-6,12-dien-1-oic acid
  38. (2R,10R,11E,16S,17E)-20-((R,E)-5-((R,E)-N-(2-Acetamidoethyl)-2,4-dimethyl-5-((3-(2-oxopyrrolidin-1-yl)propyl)amino)pent-3-enamido)-2,4-dimethylpent-3-enoyl)-1-amino-14-(3-aminopropyl)-2-(((1-(3-carboxy-4-(6-hydroxy-3-oxo-3H-xanthen-9-yl)phenyl)-2,5-dioxopyrrolidin-3-yl)thio)methyl)-10,12,16,18-tetramethyl-1,4,9,15-tetraoxo-3,8,14,20-tetraazatricosa-11,17-dien-23-oic acid
    Compound 21a (2R,10R,11E,16S,17E)-20-((R,E)-5-((R,E)-N-(2-Acetamidoethyl)-2,4-dimethyl-5-((3-(2-oxopyrrolidin-1-yl)propyl)amino)pent-3-enamido)-2,4-dimethylpent-3-enoyl)-1-amino-14-(3-aminopropyl)-2-(((1-(3-carboxy-4-(6-hydroxy-3-oxo-3H-xanthen-9-yl)phenyl)-2,5-dioxopyrrolidin-3-yl)thio)methyl)-10,12,16,18-tetramethyl-1,4,9,15-tetraoxo-3,8,14,20-tetraazatricosa-11,17-dien-23-oic acid
  39. (8R,E)-4-((S,E)-5-((R,E)-5-((R,E)-N-(3-(1H-Imidazol-1-yl)propyl)-5-((benzo[d][1,3]dioxol-5-ylmethyl)amino)-2,4-dimethylpent-3-enamido)-N-isopentyl-2,4-dimethylpent-3-enamido)-2,4-dimethylpent-3-enoyl)-23-amino-16-(4-bromophenethyl)-10-(3-isopropoxypropyl)-13-(3-methoxypropyl)-6,8-dimethyl-9,12,15,18-tetraoxo-19-(2-oxo-2-((2-oxotetrahydrofuran-3-yl)amino)ethyl)-4,10,13,16,19-pentaazatricos-6-en-1-oic acid
    Compound 22 (8R,E)-4-((S,E)-5-((R,E)-5-((R,E)-N-(3-(1H-Imidazol-1-yl)propyl)-5-((benzo[d][1,3]dioxol-5-ylmethyl)amino)-2,4-dimethylpent-3-enamido)-N-isopentyl-2,4-dimethylpent-3-enamido)-2,4-dimethylpent-3-enoyl)-23-amino-16-(4-bromophenethyl)-10-(3-isopropoxypropyl)-13-(3-methoxypropyl)-6,8-dimethyl-9,12,15,18-tetraoxo-19-(2-oxo-2-((2-oxotetrahydrofuran-3-yl)amino)ethyl)-4,10,13,16,19-pentaazatricos-6-en-1-oic acid
  40. (4E,6R,10E,12R,16E,18S)-8-(3-(1H-Imidazol-1-yl)propyl)-20-((4R,E)-5-((4-(((2R)-1-amino-3-((1-(3-carboxy-4-(6-hydroxy-3-oxo-3H-xanthen-9-yl)phenyl)-2,5-dioxopyrrolidin-3-yl)thio)-1-oxopropan-2-yl)amino)-4-oxobutyl)amino)-2,4-dimethyl-5-oxopent-2-en-1-yl)-1-(benzo[d][1,3]dioxol-5-yl)-14-isopentyl-4,6,10,12,16,18-hexamethyl-7,13,19-trioxo-2,8,14,20-tetraazatricosa-4,10,16-trien-23-oic acid
    Compound 22a (4E,6R,10E,12R,16E,18S)-8-(3-(1H-Imidazol-1-yl)propyl)-20-((4R,E)-5-((4-(((2R)-1-amino-3-((1-(3-carboxy-4-(6-hydroxy-3-oxo-3H-xanthen-9-yl)phenyl)-2,5-dioxopyrrolidin-3-yl)thio)-1-oxopropan-2-yl)amino)-4-oxobutyl)amino)-2,4-dimethyl-5-oxopent-2-en-1-yl)-1-(benzo[d][1,3]dioxol-5-yl)-14-isopentyl-4,6,10,12,16,18-hexamethyl-7,13,19-trioxo-2,8,14,20-tetraazatricosa-4,10,16-trien-23-oic acid
  41. (6E,8R,12E,14R)-4-((R,E)-5-((R,E)-N-(3-(1H-Imidazol-1-yl)propyl)-5-(isopentylamino)-2,4-dimethylpent-3-enamido)-2,4-dimethylpent-3-enoyl)-10-(2-acetamidoethyl)-29-amino-22-(4-bromophenethyl)-16-(3-isopropoxypropyl)-19-(3-methoxypropyl)-6,8,12,14-tetramethyl-9,15,18,21,24-pentaoxo-25-(2-oxo-2-((2-oxotetrahydrofuran-3-yl)amino)ethyl)-4,10,16,19,22,25-hexaazanonacosa-6,12-dien-1-oic acid
    Compound 23 (6E,8R,12E,14R)-4-((R,E)-5-((R,E)-N-(3-(1H-Imidazol-1-yl)propyl)-5-(isopentylamino)-2,4-dimethylpent-3-enamido)-2,4-dimethylpent-3-enoyl)-10-(2-acetamidoethyl)-29-amino-22-(4-bromophenethyl)-16-(3-isopropoxypropyl)-19-(3-methoxypropyl)-6,8,12,14-tetramethyl-9,15,18,21,24-pentaoxo-25-(2-oxo-2-((2-oxotetrahydrofuran-3-yl)amino)ethyl)-4,10,16,19,22,25-hexaazanonacosa-6,12-dien-1-oic acid
  42. (2R,10R,11E,16R,17E)-20-((R,E)-5-((R,E)-N-(3-(1H-Imidazol-1-yl)propyl)-5-(isopentylamino)-2,4-dimethylpent-3-enamido)-2,4-dimethylpent-3-enoyl)-14-(2-acetamidoethyl)-1-amino-2-(((1-(3-carboxy-4-(6-hydroxy-3-oxo-3H-xanthen-9-yl)phenyl)-2,5-dioxopyrrolidin-3-yl)thio)methyl)-10,12,16,18-tetramethyl-1,4,9,15-tetraoxo-3,8,14,20-tetraazatricosa-11,17-dien-23-oic acid
    Compound 23a (2R,10R,11E,16R,17E)-20-((R,E)-5-((R,E)-N-(3-(1H-Imidazol-1-yl)propyl)-5-(isopentylamino)-2,4-dimethylpent-3-enamido)-2,4-dimethylpent-3-enoyl)-14-(2-acetamidoethyl)-1-amino-2-(((1-(3-carboxy-4-(6-hydroxy-3-oxo-3H-xanthen-9-yl)phenyl)-2,5-dioxopyrrolidin-3-yl)thio)methyl)-10,12,16,18-tetramethyl-1,4,9,15-tetraoxo-3,8,14,20-tetraazatricosa-11,17-dien-23-oic acid
  43. 20-Amino-4-(2-(2-(N-(3-aminopropyl)-2-((3-aminopropyl)amino)acetamido)-N-isopentylacetamido)acetyl)-13-(4-bromophenethyl)-7-(3-isopropoxypropyl)-10-(3-methoxypropyl)-6,9,12,15-tetraoxo-16-(2-oxo-2-((2-oxotetrahydrofuran-3-yl)amino)ethyl)-4,7,10,13,16-pentaazaicosan-1-oic acid
    Compound 24 20-Amino-4-(2-(2-(N-(3-aminopropyl)-2-((3-aminopropyl)amino)acetamido)-N-isopentylacetamido)acetyl)-13-(4-bromophenethyl)-7-(3-isopropoxypropyl)-10-(3-methoxypropyl)-6,9,12,15-tetraoxo-16-(2-oxo-2-((2-oxotetrahydrofuran-3-yl)amino)ethyl)-4,7,10,13,16-pentaazaicosan-1-oic acid
  44. (2R)-1-Amino-6-(4-aminobutyl)-18-(2-(2-(N-(3-aminopropyl)-2-((3-aminopropyl)amino)acetamido)-N-isopentylacetamido)acetyl)-9-(4-bromophenethyl)-2-(((1-(3-carboxy-4-(6-hydroxy-3-oxo-3H-xanthen-9-yl)phenyl)-2,5-dioxopyrrolidin-3-yl)thio)methyl)-15-(3-isopropoxypropyl)-12-(3-methoxypropyl)-1,4,7,10,13,16-hexaoxo-3,6,9,12,15,18-hexaazahenicosan-21-oic acid
    Compound 24a (2R)-1-Amino-6-(4-aminobutyl)-18-(2-(2-(N-(3-aminopropyl)-2-((3-aminopropyl)amino)acetamido)-N-isopentylacetamido)acetyl)-9-(4-bromophenethyl)-2-(((1-(3-carboxy-4-(6-hydroxy-3-oxo-3H-xanthen-9-yl)phenyl)-2,5-dioxopyrrolidin-3-yl)thio)methyl)-15-(3-isopropoxypropyl)-12-(3-methoxypropyl)-1,4,7,10,13,16-hexaoxo-3,6,9,12,15,18-hexaazahenicosan-21-oic acid
  45. 29-Amino-22-(4-bromophenethyl)-7-isopentyl-16-(3-isopropoxypropyl)-19-(3-methoxypropyl)-6,9,12,15,18,21,24-heptaoxo-25-(2-oxo-2-((2-oxotetrahydrofuran-3-yl)amino)ethyl)-10,13-bis(3-(2-oxopyrrolidin-1-yl)propyl)-4,7,10,13,16,19,22,25-octaazanonacosan-1-oic acid
    Compound 25 29-Amino-22-(4-bromophenethyl)-7-isopentyl-16-(3-isopropoxypropyl)-19-(3-methoxypropyl)-6,9,12,15,18,21,24-heptaoxo-25-(2-oxo-2-((2-oxotetrahydrofuran-3-yl)amino)ethyl)-10,13-bis(3-(2-oxopyrrolidin-1-yl)propyl)-4,7,10,13,16,19,22,25-octaazanonacosan-1-oic acid
  46. (2R)-1-Amino-6-(4-aminobutyl)-9-(4-bromophenethyl)-2-(((1-(3-carboxy-4-(6-hydroxy-3-oxo-3H-xanthen-9-yl)phenyl)-2,5-dioxopyrrolidin-3-yl)thio)methyl)-24-isopentyl-15-(3-isopropoxypropyl)-12-(3-methoxypropyl)-1,4,7,10,13,16,19,22,25-nonaoxo-18,21-bis(3-(2-oxopyrrolidin-1-yl)propyl)-3,6,9,12,15,18,21,24,27-nonaazatriacontan-30-oic acid
    Compound 25a (2R)-1-Amino-6-(4-aminobutyl)-9-(4-bromophenethyl)-2-(((1-(3-carboxy-4-(6-hydroxy-3-oxo-3H-xanthen-9-yl)phenyl)-2,5-dioxopyrrolidin-3-yl)thio)methyl)-24-isopentyl-15-(3-isopropoxypropyl)-12-(3-methoxypropyl)-1,4,7,10,13,16,19,22,25-nonaoxo-18,21-bis(3-(2-oxopyrrolidin-1-yl)propyl)-3,6,9,12,15,18,21,24,27-nonaazatriacontan-30-oic acid
  47. 20-Amino-4-(2-(2-(2-((benzo[d][1,3]dioxol-5-ylmethyl)amino)-N-(3-(2-oxopyrrolidin-1-yl)propyl)acetamido)-N-(3-(2-oxopyrrolidin-1-yl)propyl)acetamido)acetyl)-13-(4-bromophenethyl)-7-(3-isopropoxypropyl)-10-(3-methoxypropyl)-6,9,12,15-tetraoxo-16-(2-oxo-2-((2-oxotetrahydrofuran-3-yl)amino)ethyl)-4,7,10,13,16-pentaazaicosan-1-oic acid
    Compound 26 20-Amino-4-(2-(2-(2-((benzo[d][1,3]dioxol-5-ylmethyl)amino)-N-(3-(2-oxopyrrolidin-1-yl)propyl)acetamido)-N-(3-(2-oxopyrrolidin-1-yl)propyl)acetamido)acetyl)-13-(4-bromophenethyl)-7-(3-isopropoxypropyl)-10-(3-methoxypropyl)-6,9,12,15-tetraoxo-16-(2-oxo-2-((2-oxotetrahydrofuran-3-yl)amino)ethyl)-4,7,10,13,16-pentaazaicosan-1-oic acid
  48. (2R)-1-Amino-6-(4-aminobutyl)-18-(2-(2-(2-((benzo[d][1,3]dioxol-5-ylmethyl)amino)-N-(3-(2-oxopyrrolidin-1-yl)propyl)acetamido)-N-(3-(2-oxopyrrolidin-1-yl)propyl)acetamido)acetyl)-9-(4-bromophenethyl)-2-(((1-(3-carboxy-4-(6-hydroxy-3-oxo-3H-xanthen-9-yl)phenyl)-2,5-dioxopyrrolidin-3-yl)thio)methyl)-15-(3-isopropoxypropyl)-12-(3-methoxypropyl)-1,4,7,10,13,16-hexaoxo-3,6,9,12,15,18-hexaazahenicosan-21-oic acid
    Compound 26a (2R)-1-Amino-6-(4-aminobutyl)-18-(2-(2-(2-((benzo[d][1,3]dioxol-5-ylmethyl)amino)-N-(3-(2-oxopyrrolidin-1-yl)propyl)acetamido)-N-(3-(2-oxopyrrolidin-1-yl)propyl)acetamido)acetyl)-9-(4-bromophenethyl)-2-(((1-(3-carboxy-4-(6-hydroxy-3-oxo-3H-xanthen-9-yl)phenyl)-2,5-dioxopyrrolidin-3-yl)thio)methyl)-15-(3-isopropoxypropyl)-12-(3-methoxypropyl)-1,4,7,10,13,16-hexaoxo-3,6,9,12,15,18-hexaazahenicosan-21-oic acid
  49. N-(2-Acetamidoethyl)-2-(2-(N-(2-acetamidoethyl)-2-((2-morpholinoethyl)amino)acetamido)-N-(4-methoxybenzyl)acetamido)-N-(16-amino-9-(4-bromophenethyl)-3-(3-isopropoxypropyl)-6-(3-methoxypropyl)-2,5,8,11-tetraoxo-12-(2-oxo-2-((2-oxotetrahydrofuran-3-yl)amino)ethyl)-3,6,9,12-tetraazahexadecyl)acetamide
    Compound 27 N-(2-Acetamidoethyl)-2-(2-(N-(2-acetamidoethyl)-2-((2-morpholinoethyl)amino)acetamido)-N-(4-methoxybenzyl)acetamido)-N-(16-amino-9-(4-bromophenethyl)-3-(3-isopropoxypropyl)-6-(3-methoxypropyl)-2,5,8,11-tetraoxo-12-(2-oxo-2-((2-oxotetrahydrofuran-3-yl)amino)ethyl)-3,6,9,12-tetraazahexadecyl)acetamide
  50. 5-(3-(((R)-18,24-Bis(2-acetamidoethyl)-6-(4-aminobutyl)-9-(4-bromophenethyl)-2-carbamoyl-15-(3-isopropoxypropyl)-21-(4-methoxybenzyl)-12-(3-methoxypropyl)-29-morpholino-4,7,10,13,16,19,22,25-octaoxo-3,6,9,12,15,18,21,24,27-nonaazanonacosyl)thio)-2,5-dioxopyrrolidin-1-yl)-2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)benzoic acid
    Compound 27a 5-(3-(((R)-18,24-Bis(2-acetamidoethyl)-6-(4-aminobutyl)-9-(4-bromophenethyl)-2-carbamoyl-15-(3-isopropoxypropyl)-21-(4-methoxybenzyl)-12-(3-methoxypropyl)-29-morpholino-4,7,10,13,16,19,22,25-octaoxo-3,6,9,12,15,18,21,24,27-nonaazanonacosyl)thio)-2,5-dioxopyrrolidin-1-yl)-2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)benzoic acid
  51. 2-((3-(1H-Imidazol-1-yl)propyl)amino)-N-(22-amino-3-benzyl-15-(4-bromophenethyl)-9-(3-isopropoxypropyl)-6-(4-methoxybenzyl)-12-(3-methoxypropyl)-2,5,8,11,14,17-hexaoxo-18-(2-oxo-2-((2-oxotetrahydrofuran-3-yl)amino)ethyl)-3,6,9,12,15,18-hexaazadocosyl)-N-(3-(2-oxopyrrolidin-1-yl)propyl)acetamide
    Compound 28 2-((3-(1H-Imidazol-1-yl)propyl)amino)-N-(22-amino-3-benzyl-15-(4-bromophenethyl)-9-(3-isopropoxypropyl)-6-(4-methoxybenzyl)-12-(3-methoxypropyl)-2,5,8,11,14,17-hexaoxo-18-(2-oxo-2-((2-oxotetrahydrofuran-3-yl)amino)ethyl)-3,6,9,12,15,18-hexaazadocosyl)-N-(3-(2-oxopyrrolidin-1-yl)propyl)acetamide
  52. 5-(3-(((R)-6-(4-Aminobutyl)-21-benzyl-9-(4-bromophenethyl)-2-carbamoyl-30-(1H-imidazol-1-yl)-15-(3-isopropoxypropyl)-18-(4-methoxybenzyl)-12-(3-methoxypropyl)-4,7,10,13,16,19,22,25-octaoxo-24-(3-(2-oxopyrrolidin-1-yl)propyl)-3,6,9,12,15,18,21,24,27-nonaazatriacontyl)thio)-2,5-dioxopyrrolidin-1-yl)-2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)benzoic acid
    Compound 28a 5-(3-(((R)-6-(4-Aminobutyl)-21-benzyl-9-(4-bromophenethyl)-2-carbamoyl-30-(1H-imidazol-1-yl)-15-(3-isopropoxypropyl)-18-(4-methoxybenzyl)-12-(3-methoxypropyl)-4,7,10,13,16,19,22,25-octaoxo-24-(3-(2-oxopyrrolidin-1-yl)propyl)-3,6,9,12,15,18,21,24,27-nonaazatriacontyl)thio)-2,5-dioxopyrrolidin-1-yl)-2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)benzoic acid
  53. 4-(2-(2-(N-(2-Acetamidoethyl)-2-((3-aminopropyl)amino)acetamido)-N-(3-(2-oxopyrrolidin-1-yl)propyl)acetamido)acetyl)-20-amino-13-(4-bromophenethyl)-7-(3-isopropoxypropyl)-10-(3-methoxypropyl)-6,9,12,15-tetraoxo-16-(2-oxo-2-((2-oxotetrahydrofuran-3-yl)amino)ethyl)-4,7,10,13,16-pentaazaicosan-1-oic acid
    Compound 29 4-(2-(2-(N-(2-Acetamidoethyl)-2-((3-aminopropyl)amino)acetamido)-N-(3-(2-oxopyrrolidin-1-yl)propyl)acetamido)acetyl)-20-amino-13-(4-bromophenethyl)-7-(3-isopropoxypropyl)-10-(3-methoxypropyl)-6,9,12,15-tetraoxo-16-(2-oxo-2-((2-oxotetrahydrofuran-3-yl)amino)ethyl)-4,7,10,13,16-pentaazaicosan-1-oic acid
  54. (2R)-18-(2-(2-(N-(2-Acetamidoethyl)-2-((3-aminopropyl)amino)acetamido)-N-(3-(2-oxopyrrolidin-1-yl)propyl)acetamido)acetyl)-1-amino-6-(4-aminobutyl)-9-(4-bromophenethyl)-2-(((1-(3-carboxy-4-(6-hydroxy-3-oxo-3H-xanthen-9-yl)phenyl)-2,5-dioxopyrrolidin-3-yl)thio)methyl)-15-(3-isopropoxypropyl)-12-(3-methoxypropyl)-1,4,7,10,13,16-hexaoxo-3,6,9,12,15,18-hexaazahenicosan-21-oic acid
    Compound 29a (2R)-18-(2-(2-(N-(2-Acetamidoethyl)-2-((3-aminopropyl)amino)acetamido)-N-(3-(2-oxopyrrolidin-1-yl)propyl)acetamido)acetyl)-1-amino-6-(4-aminobutyl)-9-(4-bromophenethyl)-2-(((1-(3-carboxy-4-(6-hydroxy-3-oxo-3H-xanthen-9-yl)phenyl)-2,5-dioxopyrrolidin-3-yl)thio)methyl)-15-(3-isopropoxypropyl)-12-(3-methoxypropyl)-1,4,7,10,13,16-hexaoxo-3,6,9,12,15,18-hexaazahenicosan-21-oic acid
  55. (R)-Ethyl 4-benzyl-2-oxooxazolidine-3-carboxylate
    Compound 30 (R)-Ethyl 4-benzyl-2-oxooxazolidine-3-carboxylate
  56. (R,E)-N-(2-((2-((2-((2-Amino-2-oxoethyl)(4-aminobutyl)amino)-2-oxoethyl)(3-methoxypropyl)amino)-2-oxoethyl)(benzyl)amino)-2-oxoethyl)-N-benzyl-5-(N-benzyl-2-(N-benzyl-2-(benzylamino)acetamido)acetamido)-2,4-dimethylpent-3-enamide
    Compound 31 (R,E)-N-(2-((2-((2-((2-Amino-2-oxoethyl)(4-aminobutyl)amino)-2-oxoethyl)(3-methoxypropyl)amino)-2-oxoethyl)(benzyl)amino)-2-oxoethyl)-N-benzyl-5-(N-benzyl-2-(N-benzyl-2-(benzylamino)acetamido)acetamido)-2,4-dimethylpent-3-enamide
  57. (2R,E)-N-(3-(1H-Imidazol-1-yl)propyl)-N-((13R,14E,19S,20E)-1-amino-17-benzyl-8-(3-isopropoxypropyl)-13,15,19,21-tetramethyl-6,9,12,18-tetraoxo-5-(2-oxo-2-((2-oxotetrahydrofuran-3-yl)amino)ethyl)-11-phenethyl-5,8,11,17-tetraazadocosa-14,20-dien-22-yl)-5-((S,E)-5-((4-bromobenzyl)amino)-2,4-dimethyl-N-phenethylpent-3-enamido)-2,4-dimethylpent-3-enamide
    Compound 32 (2R,E)-N-(3-(1H-Imidazol-1-yl)propyl)-N-((13R,14E,19S,20E)-1-amino-17-benzyl-8-(3-isopropoxypropyl)-13,15,19,21-tetramethyl-6,9,12,18-tetraoxo-5-(2-oxo-2-((2-oxotetrahydrofuran-3-yl)amino)ethyl)-11-phenethyl-5,8,11,17-tetraazadocosa-14,20-dien-22-yl)-5-((S,E)-5-((4-bromobenzyl)amino)-2,4-dimethyl-N-phenethylpent-3-enamido)-2,4-dimethylpent-3-enamide
  58. (S,E)-N-((S,E)-5-Amino-2,4-dimethyl-5-oxopent-2-en-1-yl)-5-((S,E)-N-(benzo[d][1,3]dioxol-5-ylmethyl)-5-((R,E)-N-(benzo[d][1,3]dioxol-5-ylmethyl)-5-((benzo[d][1,3]dioxol-5-ylmethyl)amino)-2,4-dimethylpent-3-enamido)-2,4-dimethylpent-3-enamido)-N-benzyl-2,4-dimethylpent-3-enamide
    Compound 53 (S,E)-N-((S,E)-5-Amino-2,4-dimethyl-5-oxopent-2-en-1-yl)-5-((S,E)-N-(benzo[d][1,3]dioxol-5-ylmethyl)-5-((R,E)-N-(benzo[d][1,3]dioxol-5-ylmethyl)-5-((benzo[d][1,3]dioxol-5-ylmethyl)amino)-2,4-dimethylpent-3-enamido)-2,4-dimethylpent-3-enamido)-N-benzyl-2,4-dimethylpent-3-enamide
  59. 5-(3-(((R)-1-(Benzo[d][1,3]dioxol-5-yl)-5,8-bis(benzo[d][1,3]dioxol-5-ylmethyl)-11-benzyl-20-carbamoyl-4,7,10,13,18-pentaoxo-2,5,8,11,14,19-hexaazahenicosan-21-yl)thio)-2,5-dioxopyrrolidin-1-yl)-2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)benzoic acid
    Compound 54 5-(3-(((R)-1-(Benzo[d][1,3]dioxol-5-yl)-5,8-bis(benzo[d][1,3]dioxol-5-ylmethyl)-11-benzyl-20-carbamoyl-4,7,10,13,18-pentaoxo-2,5,8,11,14,19-hexaazahenicosan-21-yl)thio)-2,5-dioxopyrrolidin-1-yl)-2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)benzoic acid

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Author information

Affiliations

  1. Department of Chemistry, The Scripps Research Institute, Scripps Florida, Jupiter, Florida 33458, USA

    • Claudio Aquino,
    • Mohosin Sarkar,
    • Kimberly Mendes,
    • Thomas Kodadek &
    • Glenn C. Micalizio
  2. Department of Cancer Biology, The Scripps Research Institute, Scripps Florida, Jupiter, Florida 33458, USA

    • Mohosin Sarkar,
    • Kimberly Mendes &
    • Thomas Kodadek
  3. Department of Molecular Therapeutics, The Scripps Research Institute, Scripps Florida, Jupiter, Florida 33458, USA

    • Michael J. Chalmers

Contributions

G.C.M. and T.K. conceived and directed the project. C.A. and M.S. contributed equally to the execution of this work. C.A. prepared acid 1 and conducted all solution-phase chemistry. M.S. and C.A. conducted all solid-phase experiments, library synthesis, decoding experiments and biochemical evaluation of the hits. M.S. purified the p53–DBD, which was cloned and expressed by K.M., and conducted the biochemical characterization of the hit. M.C. developed the ETD-based method for compound decoding. G.C.M. and T.K. wrote the manuscript.

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The authors declare no competing financial interests.

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