Assembly of Complex Macrocycles by Incrementally Amalgamating Unprotected Peptides with a Designed Four-Armed Insert

Article abstract of DOI:10.1021/acs.joc.7b02958

We describe the asymmetric synthesis of a highly substituted ω-octynoic acid derivative and demonstrate its utility for generating complex macrocycles from unprotected peptides. The molecule harbors an isolated quaternary center that displays four uniquely functionalized arms, each of which can be reacted orthogonally in sequence as the molecule is integrated into peptide structure. These processing sequences entail (1) scaffold ligation, (2) macrocyclization via internal aromatic alkylations or catalyzed etherifications, (3) acyliminium ion mediated embedding of condensed heterocycles, and (4) terminal alkyne derivatization or dimerization reactions. Numerous polycycles are prepared and fully characterized in this study. Factors that influence reaction efficiencies and selectivity are also probed. We construct a novel mimic of the second mitochondria derived activator of caspase using these techniques, wherein subtle variations in macrocycle connectivity have a marked impact on performance. In general, the chemistry is an important step toward facile, systematic access to complex peptidomimetics synthesized by directly altering the structure and properties of machine-made oligomers.

Full text of DOI:10.1021/acs.joc.7b02958

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Article
Assembly of complex macrocycles by incrementally amalgamating
unprotected peptides with a designed four-armed insert
Brice H. Curtin, Francesco Manoni, Jiyong Park, Luke J Sisto, Yu-
hong Lam, Michel Gravel, Anne Roulston, and Patrick G Harran
J. Org. Chem., Just Accepted Manuscript • DOI: 10.1021/acs.joc.7b02958 • Publication Date (Web): 01 Mar 2018
Downloaded from http://pubs.acs.org on March 2, 2018
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The Journal of Organic Chemistry
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Assembly of complex macrocycles by incrementally amalgamating
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Brice H. Curtin, Francesco Manoni, Jiyong Park, Luke J. Sisto, Yu-hong Lam, Michel Gravel,
‡
†
Anne Roulston, Patrick G. Harran*
†
Department of Chemistry and Biochemistry, University of California Los Angeles, Los Angeles, California 90095, USA.
Laboratory for Therapeutic Development, Rosalind and Morris Goodman Cancer Research Centre and Department of Bio-
chemistry, McGill University, Montreal, Quebec H3G 1Y6, Canada.
‡
ABSTRACT: We describe the asymmetric synthesis of a highly-substituted ω-octynoic acid derivative and demonstrate its utility
for generating complex macrocycles from unprotected peptides. The molecule harbors an isolated quaternary center that displays
four uniquely functionalized arms, each of which can be reacted orthogonally in sequence as the molecule is integrated into peptide
structure. These processing sequences entail 1) scaffold ligation 2) macrocyclization via internal aromatic alkylations or catalyzed
etherifications 3) acyliminium ion mediated embedding of condensed heterocycles and 4) terminal alkyne derivatization or dimeri-
zation reactions. Numerous polycycles are prepared and fully characterized in this study. Factors that influence reaction efficiencies
and selectivity are also probed. We construct a novel mimic of the second mitochondria derived activator of caspase using these
techniques, wherein subtle variations in macrocycle connectivity have a marked impact on performance. In general, the chemistry is
an important step towards facile, systematic access to complex peptidomimetics synthesized by directly altering the structure and
properties of machine-made oligomers.
INTRODUCTION
Macrocycles have the potential to expand the scope of drug
discovery due to their ability to scaffold extended pharmaco-
1
–5
phores. This attribute is thought to facilitate interactions
with protein surfaces, including those involved in biological
signaling pathways. Signaling events mediated by protein-
protein interactions (PPIs) provide the largest class of potential
drug targets, but these are difficult to engage with small mole-
cules. That said, numerous PPIs are mediated by a short
peptide sequence in one partner, and that sequence therefore
is a logical starting point for drug discovery. However, poor
transport properties and limited in vivo stability of small pep-
6
,7
5
8
,9
5,10
tides are perennial challenges to this approach.
wards improved performance involves cyclization, as cyclic
peptides can possess improved cellular permeability and sta-
A step to-
1
1,12
bility relative to acyclic counterparts.
Conformational re-
striction affords defined topologies that can shield polar sur-
face area while maintaining a desired binding motif. Numer-
ous methods exist to form cyclic peptides, including conven-
tional amidations as well as a range of alternate constructions
developed more recently.
are both general and high yielding. Cyclic peptides can also be
made in enormous numbers using nucleic acid encoding tech-
The question is whether ring formation alone can
sufficiently alter properties to afford valuable lead structures.
1
3–23
Many of these creative methods
24–26
nologies.
2
7–30
While in certain instances the answer is yes,
we have been
working under the assumption that, in general, it is not. To
pursue further alterations in context of large ring structures,
we have designed synthetic inserts that react incrementally
with linear peptides to form macrocyclic composites. The goal
is for the hybrid molecules to retain molecular recognition
elements in the biopolymer while displaying that functionality
Figure 1. Molecular amalgamations made possible by increasing-
ly capable scaffolding reagents. Reagents 1, 2, and (+)-5 engage
unprotected peptides in short processing sequences that generate
complex products having defined shapes and altered properties.
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as part of stable, amphipathic polycycles having defined lem. Initially we considered elaborating intermediates used to
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shapes and improved pharmacological properties.
The
prepare 2. Because this would have further lengthened a nine-
step sequence, it was not attractive. Attention turned instead to
de novo synthesis of 5. Several tactics to generate this com-
pound seemed plausible, including asymmetric conjugate ad-
dition of a formyl anion synthon to acrylic esters 8 (Fig. 2).
Racemic intermediates could be prepared by adding nitrome-
thane to 8, but, in our hands, optically active products were
elusive. Fortunately, ideas evolved quickly from 8. Instead of
adding a one-carbon nucleophile to 8, we examined reacting a
two-carbon electrophile with 9. This involved attempts at cata-
scaffolding in our chemistry has evolved from a core cinnamyl
alcohol motif. We have shown how this unit can support large
ring forming reactions by exploiting the cinnamyl cation; gen-
erated either as a solvated ion pair under acidic conditions or
as a metal stabilized complex. The latter allows us to synthe-
size macrocyclic ethers, amines and lactones while the former
permits unique macrocyclizations via direct carbon-carbon
bonding (e.g. 3, Fig. 1A). In neither instance are protecting
groups required on the peptide. As our scaffolding has become
more functionalized, we have been able to sequence additional
reactions with macrocyclization. For example, when cinnamyl
carbonate containing propionic acid derivative 2 is used to
acylate a pyrrolic derivative of Dap-Thr-Tyr, mild acidolysis
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lyzing addition of an enol to nitroethylene. This also proved
challenging, but working with aldehyde 9 we recognized the
enantiotopic faces of its capped enol (10, P = TBS) might be
3
8
discriminated via asymmetric cyclopropanation. This could
generate an intermediate oxygenated cyclopropyl carboxylate
(11), whose fragmentation (as shown) was expected to be fac-
ile. At about this same time, Marek et al. reported that directed
carbometalation of chiral cyclopropenyl carboxylates followed
by in situ oxidation gave optically active 4-oxobutyrate deriva-
(
aq. AcOH) of the product converts the N-terminus into a pyr-
35,36
rolopiperazine via N-acyliminium ion cyclization.
Subse-
quent exposure to MsOH in MeNO
Crafts alkylation to afford a single regioisomeric macrocycle
4) in high yield. The secondary alcohol, primary carbox-
2
initiates internal Friedel-
(
39
amide, phenol and disubstituted pyrrole are unaffected in the
reaction, which occurs within minutes at room temperature.
tives directly. These reactions proceeded by way of species
analogous to 11 and suggested a clear path to enantioenriched
5
beginning with cyclopropene 12 and an appropriate organo-
Holding 2 constant, many variants of this three-step se-
quence can be executed on a range of functionalized oligo-
mers. The operations are rapid and facile. We are currently
generating numerous complex peptidomimetics in this man-
ner. At the same time, our scaffold designs continue to evolve.
We felt the methine hydrogen in 2 (namely C2-H) presented
another opportunity. By substituting this position, not only
would configurational stability at C2 be assured, we could also
explore an entirely new series of experiments. If the fourth
branch exhibited reactivity orthogonal to the other three, we
could probe whether the initially formed macrocycles could be
tagged, transannulated, or multimerized in sequence. The
structures generated by such processes have little precedent
and would position us to study their properties in detail. Im-
portantly, this could include direct comparisons to products
derived from manipulating individual peptide sequences with
1 and 2. Here we describe important steps in this direction.
metallic.
RESULTS & DISCUSSION
m-Bromophenyl propyne was synthesized from commercial
3
-bromobenzyl bromide and trimethylsilyl acetylene using a
40
Negishi protocol (Scheme 1). Reaction of this material with
ethyl diazoacetate in the presence of Corey’s trisimidazoli-
dinone dirhodium complex 16 (0.25 mol%) afforded cyclo-
41
propene carboxylate (+)-17 in 80% yield and 95% e.e. as
judged by chiral supercritical fluid chromatography. S stereo-
chemistry in this material was tentatively assigned by analogy
to Corey’s precedent, and later corroborated by NMR analyses
of diastereomeric derivatives of downstream product (+)-5
(
vide infra).
With 17 in hand, we next examined copper-mediated car-
bometalation of its strained alkene. Procedures involving Gri-
gnard 18 and catalytic amounts of copper salts were not pro-
ductive in our hands. However, when 17 was added slowly to
superstoichiometric amounts of 18 (freshly prepared, 1.4 M in
THF) and CuI at -40˚C, stirring for 30 min followed by
Based on results from model systems, we chose reagent 5
(Fig. 1B) as a target. This material retains all capabilities of 2
while adding a normal pentynyl chain at its lone chiral center.
The alkyne was anticipated to be inert to macrocyclization and
heteroannulation conditions used to react other functional
groups in the molecule. It would also provide varied options
for manipulating the resultant macrocycles. Before this idea
could be tested, however, we faced a difficult synthetic prob-
4 4
quench with premixed NH Cl/NH OH cleanly generated cy-
clopropane 19 (R = 4-pentynyl, M = H). Diastereoselectivity
appeared high and 2D-NOESY spectra of the material were
consistent with the major isomer being that drawn (see SI).
Sequencing the carbometalation with in situ oxidation was
more challenging. Among the variety of oxidants examined,
3
9,42
only lithium t-butyl peroxide proved effective.
An opti-
mized protocol involved a pre-formed organocopper species
prepared from 18 and CuI•TMEDA complex being used to
carbometalate 17. Temperature control while adding 17 was
important such that organometallic species were largely dis-
solved at -40 ºC in THF. This was critical for scalability. Un-
der such conditions, cyclopropene (+)-17 was consumed with-
in 30 min, whereupon the mixture was cooled to -78 ºC and
carefully treated with anhydrous t-BuOOLi. After aqueous
workup, aldehyde 21 was isolated directly, ostensibly via in
situ fragmentation of a transient cyclopropanoxide (i.e. 20).
Compound 21 was difficult to purify without loss, and there-
fore crude material was treated with triethylorthoformate in
the presence of catalytic TsOH. The resultant acetal was cross
Figure 2. Routes contemplated to build a highly functionalized
chiral quaternary center.
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Scheme 1. Enantioselective Synthesis of (+)-5
The Journal of Organic Chemistry
Scheme 2. Corroborating the Stereochemical Assignment
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of (+)-5
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Reaction conditions: a) 1.0 eq. (+)-5, 1.5 eq. 25, iPr NEt, DMF. b)
4:1 AcOH/H O, 22 ºC, 12 h, 51% yield over two steps. Inset:
2
partial 2-D NOESY spectrum of 27 showing key correlations
involving H21.
earlier assignment of stereochemistry in 17 made by analogy
to Corey’s results.
Initial macrocyclizations with this new scaffold were tested
in the absence of the alkyne. An analog of (+)-5 was prepared
wherein the C3 pentynyl group was replaced by methyl (by
2
Reaction conditions: a) 0.2 mol% Pd(DPEPhos)Cl , 1.4 eq. 14,
THF, 22 ºC. b) 2.0 eq. AcOH, 2.0 eq. TBAF, THF, 22 ºC, 85%
yield over two steps. c) 3.0 eq. 15, 1 eq. ethyl diazoacetate, 0.25
mol% 16, DCM, 80% yield, 95% e.e. d) 2.0 eq. 18, 2.2 eq.
TMEDA, 2.0 eq. CuI,; e) then 2.0 eq. tBuOOLi; 2:1
3
substituting CH MgBr for 18 in Scheme 1, see Fig. S2). Acyl-
ating Trp-Glu-Tyr with this molecule gave 28. Exposure of 28
to aqueous acetic acid gave Pictet-Spengler product 29 in high
yield (Scheme 3). Interestingly, all attempts to macrocyclize
this molecule by activating the cinnamyl carbonate under con-
NH
mol% Pd(PPh
reflux, 29% yield over three steps. h) 2.5 eq. TBAF, THF, 0 ºC. i)
10 eq. KOH, 2:1 EtOH/H O, 50 ºC. j) 4.5 eq. N-
4
Cl/NH
4
OH. f) 10 mol% pTSA, 3.0 eq. (EtO)
3
CH, EtOH. g) 1
3
3–35
3
)
4
, 1.2 eq. 22, 3.0 eq. Na CO , 5:1 dioxane/H
2
3
2
O,
ditions developed previously led to decomposition.
versing the order of events solved this problem. Treatment of
8 with 5 mol% Pd(PPh in DMF at room temperature initi-
Re-
2
2
3 4
)
methylmorpholine, 2.1 eq. i-BuOCOCl, DCM, -5 ºC; then 2.0 eq.
N-hydroxysuccinimide, -5 ºC to 22 ºC, 12h, 52% over three steps,
ated a high yielding cycloetherification. When that product
was dissolved in aqueous acetic acid, it slowly converted to
polycycle 30 (90% conversion after 4 days at 25 ºC). Follow-
ing preparative HPLC, analytically pure β-carboline 30 was
isolated in 28% yield (three steps from S5). The 2-D NOESY
spectrum showed correlations very similar to those observed
in 27 (Scheme 2) and stereochemistry in 30 was thus assigned
similarly. A possible explanation for the reluctance of 29 to
participate in macrocyclization reactions was the conforma-
tional restriction imposed by the Pictet-Spengler process. It
oriented the C17 and C11 β branches anti off of a rigid tetra-
hydroindolizinone core. The effect was marked. Substrate 31
was synthesized and harbored the same residues as 28, but
wherein an aminohexanoic acid spacer was inserted in be-
tween P1 and P2. Treatment with aqueous acetic acid smooth-
ly initiated a Pictet-Spengler cyclization, but the system re-
mained reluctant to macrocyclize. Only palladium-catalyzed
cycloetherification was successful, and in that case product 32
was relatively unstable, presumably due to torsional strain
present in the macrocycle. Attenuating this strain had a posi-
tive impact. Substrate 33 was synthesized. This molecule was
the same as 28 except the tryptophan residue was D-
configured. Acetic acid promoted Pictet-Spengler reaction
within this molecule occurred cleanly and subsequent macro-
cyclizations were now facile. Treatment with either TFA in
9
4% e.e. EDA= ethyl diazoacetate; NHS= N-hydroxysuccinimide.
coupled with vinyl boronate 22 (See SI) using palladium ca-
talysis to afford stable product 23 in 29% overall yield from
17. Desilylation and saponification then afforded hydroxy acid
24, which was reacted with excess iso-butyl chloroformate.
The doubly acylated species formed in situ was partially de-
composed with N-hydroxysuccinimide to afford target (+)-5 in
5
2% isolated yield. This concise route to (+)-5 gave access to
our first four-armed scaffolding reagent on multi-gram scales.
Before exploring its utility, we further probed stereochem-
istry in (+)-5. The molecule was reacted with L-tryptophan
carboxamide to give 26. When that substance was dissolved in
8
0% aqueous AcOH, its acetal quickly (< 1 hr) decomposed to
a mixture of diastereomeric hydroxy lactams. These gradually
converted to pyrrolo-β-carboline 27 (via 26a) over the next 12
1
hours. A single isomer of 27 was observed by H NMR and
HPLC. Its 2D-NOESY spectrum showed clear correlations
between the carboxamide protons H37,37’ and the C21 me-
thine hydrogen, which in turn correlated with methylene sig-
nals H16 and H17. Literature precedent and our own studies
indicated the configuration at C35 would dictate stereochemis-
3
5,36,43,44
try at C21.
If true, the relative stereochemical relation-
ships implied from NOE correlations would translate to (+)-5
being S-configured, which was further consistent with the
CH
3 2 3 4
NO or 5 mol% Pd(PPh ) in DMF gave regioisomeric
macrocycles 34 and 35 in 60% and 29% yield, respectively.
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Scheme 3. Probing Initial Macrocyclizations with a Three- Having established the new scaffold frame supported mac-
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Armed Model
rocyclizations, we turned to 5 and experiments to test the in-
ertness of its alkyne to both palladium catalysis and acidolysis
conditions used for large ring formations (Fig. 3). Acylation of
D-Trp-Glu-Tyr with (+)-5 and subsequent treatment with
3 2
aqueous acetic acid followed by TFA in CH NO (5 vol %)
gave macrocycle 36 in 31% isolated yield over three steps.
NOE correlations in 36 paralleled those observed in 27 and 34
and were fully consistent with the relative stereochemistry
drawn. As we hoped, no products derived from reactions at the
alkyne were detected, nor did the carboxylic acid or primary
carboxamide interfere. The same three-step sequence begin-
ning with 5 was repeated with D-Trp-Gln-Tyr, Pro-Ala-Lys(D-
Trp)-Tyr, and D-Trp-Glu(tyramide) to yield macrocycles 37–
39 in good per step average yields over three steps. We next
prepared the O-linked regioisomer of 39 (namely 40) by
changing the third step in the processing sequence. Instead of
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treating with TFA in MeNO
2
, the Pictet-Spengler product was
exposed to 4 mol% allyl palladium chloride dimer, 10 mol%
34
2 3
Xantphos, and stoichiometric Cs CO in DMF. The alkyne
was again unaffected. The dipeptide D-Trp-AAP* having its
C-terminus condensed with tyramine was readily processed
with 5 to afford polycycle 41. Both the alkyne and the primary
azide were unaffected, opening the possibility for transannula-
tions via Huisgen cycloadditions should that be desired in
future iterations.
Each stage of engagement with 5 was designed to be flexi-
ble. Consistent with Meldal’s results, the N-acyl iminium ion
intermediates would react with a range of proximal π-basic
3
5,36,43
aromatics.
For
example,
when
D-3-MeOPhe-
Thr(tyramide) was N-acylated with 5 and treated with aqueous
acetic acid, two isomeric products (1:1) were formed. They
were tentatively assigned as epimeric dihydropyrroloimidazole
44
diones, although alternative structures could not be ruled out.
When those materials were treated with 5 vol % TFA in
CH NO , Pictet-Spengler reaction and Friedel-Crafts macro-
3 2
cyclization occurred concomitantly to afford a single macro-
cyclization product (42) in good overall yield. Neither the
alkyne nor the secondary alcohol were affected. In the case of
D-Trp(5Br)-His(tyramide), its reaction with 5 gave a product
that resisted Pictet-Spengler reaction in aqueous AcOH. How-
ever, addition of 10 vol % H PO caused rapid cyclization.
3 4
Notably, without degrading the cinnamyl carbonate. The
product was then converted to macrocycle 43 by exposure to
TFA in MeNO . Alternatively, palladium-catalyzed cyclo-
2
etherifcation afforded macrocyclic cinnamyl ether 44. The
alkyne and the unprotected imidazole ring were unaffected by
either process.
2
Reaction conditions: a) 4:1 AcOH/H O, 22 ºC, 12 hours. b) 2.0
eq. Cs
vol% TFA, CH
days. Note: yields quoted throughout reflect analytically pure
2
CO
3
, 5 mol% Pd(PPh
3
)
4
, DMSO, 10 mM, 4 hours. c) 5
Lastly, in the course of these studies, we discovered what
we believe is a unique macrocyclization process. When D-Trp-
Cys(St-Bu)(tyramide) was N-acylated with (+)-5 and the
product was dissolved in aq. AcOH, a Pictet-Spengler reaction
occurred uneventfully. However, when that molecule was
3
NO , 5 mM, 2 hours. d) 4:1 AcOH/H
2
2
O, 22 ºC, 4
2
material isolated (prep-HPLC or SiO chromatography) after full
sequence beginning with (+)-S5.
Efficient macrocyclization via either internal Friedel-Crafts
alkylation or Tsuji-Trost cinnamylation was consistent with
earlier studies (e.g. Fig 1A) and reflective of the relative stere-
ochemistry in the Pictet-Spengler product now positioning
reactive termini syn, thereby favoring ring closures. The use of
a D-configured P1 residue in conjunction with (+)-5 to permit
syntheses of unique structures such as 34 and 35 was an excel-
lent outcome. It should be noted, however, the same result
could in principle be achieved using all L-configured amino
acids and the enantiomer of 5.
2
treated with TFA in MeNO , two products (~1:1) formed in
good yield. One was the internal Friedel-Crafts alkylation
product 45a, as expected. The second lacked a tert-butyl group
and its spectroscopic data were consistent with allylic disulfide
containing macrocycle 45b. This outcome was interpreted in
terms of a solvated cinnamyl cation being captured by the
distal sulfur of the disulfide and the incipient sulfonium ion
extruding isobutylene. Macrocyclic allylic disulfides of this
type may be manipulated in a host of ways by partial oxygena-
tion reactions and/or sigmatropic rearrangements of derived
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8
9
0
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2
3
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5
6
7
8
9
0
Figure 3. Macrocyclic products obtained by acylation of unprotected peptides with (+)-5, followed by N-acyliminium ion cyclization, and
either acid-mediated Friedel-Crafts alkylation or palladium-catalyzed macrocycloetherification. Note: for yield calculations see Scheme 3.
ylides. Towards this end, experiments to test the generality
and efficiency of the macrocyclization reaction, both in the
presence and absence of competing internal nucleophiles are
ongoing.
tides and serve as a linker site for assembly of antibody and/or
protein drug conjugates.
A major goal for this program is to allow biologically active
peptides to be molded directly into potent and stable lead
In all processing sequences using (+)-5 to date, the alkyne
has been inert to chemistries used to synthesize macrocycles
having imbedded condensed heterocycles. For the eleven
structures depicted in Figure 3, and more, it was possible to
integrate (+)-5 into unprotected peptides using simple, tele-
scoped three-step sequences followed by mass-guided prepara-
tive HPLC. Pure products were routinely isolated on tens of
milligrams scales without incident.
Scheme 4. Glycoconjugation via a Copper-Catalyzed Huis-
gen Cycloaddition
While inert enough to be valuable in our schemes, the al-
kyne was certainly a handle for further manipulations. Many
natural products are glycosylated and this feature can marked-
ly alter solubility and transport properties, relative to the agly-
con, in biological systems. Viewing our composite macrocy-
cles as analogous to non-ribosomal peptides, a ready means to
add sugars to these compounds was desirable. The alkyne
made this trivial. For example, mixing polycycle 36 with
commercial 1-azido-1-deoxy-β-D-glucopyranoside in the pres-
ence of catalytic copper iodide and triethylamine proceeded
well to give unique glycoconjugate 46. Several additional
examples of this Huisgen cycloaddition are detailed in the
supporting information. We have begun to examine passive
membrane permeability in this series. Conventional wisdom
suggests molecules of this type will have difficulty entering
cells. We are interested in understanding this behavior deeply
enough that we might eventually use our templates to facilitate
permeability where it would otherwise not exist. In one of our
first Caco-2 monolayer screens, compound 46 stood out as a
substance displaying some passive permeability (see SI Table
S2). While it is minimal relative to positive controls, the fact
we observed any permeability for a molecule having as much
exposed polar surface area as 46 is striking. Collaborations to
explore structure / permeability relationships of peptidomimet-
ic macrocycles in detail are ongoing. Alkyne functionaliza-
tions will greatly aid these studies, and we note that the triple
bond may also be used for conjugation to cell penetrating pep-
Reaction Conditions: a) 1.5 eq. azido sugar, 2.5 eq Et
CuI, DMF, 22 ºC.
3
N, 10 mol%
structures for further research. To initially demonstrate the
potential of (+)-5 in this context, we began with a familiar
system. The second mitochondria derived activator of caspase
(Smac) is a homodimeric protein secreted from mitochondria
4
5
during programmed cell death. Cytoplasmic Smac relieves
inhibitor-of-apoptosis protein (IAP) mediated suppression of
caspase activity. It binds avidly to X-chromosome encoded
IAP, cellular IAP1 and cellular IAP2, and synergizes with
both TRAIL and TNFα to potently induce caspase activation
4
5
and apoptosis in human cancer cells. Smac exploits a con-
served tetrapeptide (AVPI) at its N-terminus to bind BIR do-
46
mains within IAPs. We had used traditional medicinal chem-
istry techniques earlier to develop a bivalent small molecule
4
7
mimic of Smac. That exercise went on to drive much re-
4
8,49
search as well as clinical development programs.
However,
it required several years of experiments. We were interested if
the use of (+)-5 might be able to generate Smac mimetic leads
more quickly.
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Figure 4. (A) Synthesis of macrocyclic Smac mimetic monomers. Reaction conditions: a) 1.0 eq. (+)-5, 1.5 eq. 47, iPr NEt, DMF. b) 4:1
AcOH/H O, 22 ºC, 12 h. c) 1:1 TFA/TFE, 5 mM in substrate, 22 ºC, 7h, 13% yield (49a) and 21% yield (49b). (B) Overlay of energy-
2
minimized (B3LYP-D3) conformers of 49a (gray) & 49b (green), which orient their peptidyl segments differently within the composite
structures.
In terms of caspase inhibition, it was known from in vitro
peptide screens that the P2 position of AVPI was tolerant of
side chain variations, and that an aromatic residue was pre-
although the relevance of this analysis to bound states was as
yet unclear.
Despite structural homology, slight differences in BIR do-
4
7,50–52
ferred at P4.
We therefore prepared peptide 47, wherein
main structures within IAPs have been leveraged to design
the P1 and P3 positions were unaltered. The P2 position was
occupied by a glutamic acid derivative that provided both an
attachment point for (+)-5 and a means to generate an N-
acyliminium ion from the composite. Lastly, L-O-phenyl phe-
nylalaninol was placed at P4 such that it could participate in
alkylative macrocyclizations while also displaying an aromatic
side chain.
53,54
cIAP-selective antagonists.
Because XIAP, cIAP1, and
cIAP2 function independently, and differently, to block apop-
tosis, selective antagonists have been coveted as research
tools.
Smac protein exists as a native dimer and, in the case of
XIAP, binds simultaneously to adjacent BIR domains within
its structure. We had exploited this previously by dimerizing
monomeric BIR3 domain ligands, thereby achieving excep-
Treatment of 47 with (+)-5 gave 48 in 71% yield and with-
out competitive acylation of the N-terminus. Hydrolysis in
aqueous acetic acid then generated hydroxy lactam intermedi-
ates. These species were concentrated to dryness, re-dissolved
in trifluoroethanol and treated with TFA (1:1 final, 5 mM in
substrate) at 25 ºC. This promoted an N-acyliminium ion cy-
clization and concomitant macrocyclization. The original ex-
pectation was that ion 48a (Fig. 4) would be trapped by the
adjacent amide to form a diacyl imidazolidine (e.g. 49c, Fig.
4A). However, extensive NMR analyses (including HMBC
and NOESY spectra) of the two isolated products showed
them to be regioisomeric tetrahydroindenopyrrolones 49a and
4
7
tional Smac mimicry. Anticipating similar behavior, we oxi-
datively dimerized 49a and 49b via Glaser coupling. This in-
volved treating their free-base forms with Cu(OAc)
2
and piper-
idine in 1:1 CH CN/MeOH at 70 ºC (Fig. 5A). Symmetric
3
diynes 50 and 6 were isolated in 40% and 60% yields, respec-
tively. Avidities for recombinant XIAP, cIAP1, and cIAP2
(
BIR2-BIR3 domain constructs) were then evaluated by com-
petitive binding using a fluorescence polarization (FP) assay
Fig. 5B). The same fluorescein labeled dimeric Smac peptide
(
FP probe was employed in all experiments (see SI Fig. S5).
Tetralogic’s clinical compound Birinapant™ was used as a
4
9b. Similar to logic invoked for 42 (vide supra), this outcome
was rationalized in terms of a transient diacyl imidazolidine
49c) giving way to more stable C-C bonded products via in-
55
positive control. Linear peptide 47 weakly displaced the FP
probe from all three IAP constructs, although it did discrimi-
nate cIAP1 from cIAP2. Macrocyclic monomer 49a was com-
parable to its precursor 47, but macrocycle 49b was not. As
(
ternal Friedel-Crafts alkylation. The closest aromatic ring to
ion 48a was that of the scaffold, and therefore 49a/b were
formed. To our knowledge, these macrocycles are without
precedent. Moreover, from (+)-5, they were prepared and puri-
fied in less than 48 hours.
5
6
expected for a monomer, it remained a poor competitor for
XIAP, but it displaced the FP probe from cIAPs with low na-
nomolar efficacy. In fact, it was 13 times more effective than
4
9a against both cIAP1 and cIAP2. This highlights a phenom-
We were now positioned to study how subtle differences in
ring connectivity might affect IAP binding and domain selec-
tivity. In silico geometry optimization and conformational
searches suggested that 49a and b would display their peptide
regions differently (Fig. 4B, see SI for computational details),
ena we did not fully appreciate, yet one that may be general
for molecules of this type. Namely, that subtle variations in
macrocycle topology and pharmacophore display can marked-
ly alter performance. Macrocyclic monomer 49b also
showed excellent cIAP1:XIAP selectivity (>250:1). The abil-
5
7
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Figure 5. (A) Dimerization of macrocyclic monomers. Reaction Conditions: a) 1.0 eq. 49a or 49b, 7.0 eq. Cu(OAc) , 7.0 eq. piperidine,
1
3
:1 MeOH/CH CN, 40% yield (50) or 60% yield (6). (B) Fluorescence polarization assay for competitive displacement of a labeled biva-
lent Smac-mimetic peptide from recombinant Bir2-Bir3 constructs of indicated IAP protein. Data is reported as IC50 values in nM (average
of 2 biological and 2 technical replicates). (C) & (D) Final snapshots of 100 ns MD simulations of 49a and 49b bound to the BIR3 domain
of cIAP1, respectively. Intermolecular hydrogen bonds are indicated in yellow (enlarged to show detail in SI Fig. S10).
ity of dimeric compounds 50 & 6 to compete for cIAP1/2
binding was similar to their respective monomers, a finding
consistent with the 1:2 binding stoichiometry of Smac protein
to these particular IAPs. Dimer 6, on the other hand, was the
only compound to show competitive binding to XIAP, while
remaining able to potently displace the FP probe from cIAPs,
especially cIAP1. The data above reflects competition IC50
values rather than direct binding constants although, for com-
parison, Birinipant is reported to bind to full length XIAP and
of-principle exercise in a well-characterized system, we be-
lieve the chemistry has broad potential to create and optimize
complex antagonists of protein-protein interactions, especially
those mediated by a short-linear-interacting-motif (SLIM) in
9
one partner.
CONCLUSION
We have developed a short, scalable and enantioselective
synthesis of our first four-armed scaffolding reagent. This
molecule can be incrementally integrated into a range of oli-
gomeric substrates, wherein the composite products are stable
polycycles having defined conformations. By varying the or-
der of events, ring forming modalities, and derivatization
schemes, countless new complex structures are potentially
available. From such collections the search for islands of use-
ful pharmacological properties can proceed in ways not possi-
ble previously with increasingly intricate structures being
made using multiple template generations (i.e. 1, 2, & (+)-5).
Scaffold design and utilization within the project is continually
advancing, and attempts to exploit the alkyne (and its homo-
logs) in (+)-5 for novel transannulation reactions are ongoing.
Bridged macrocycles anticipated from those studies could
bring yet another novel compound class into consideration.
5
5
D
cIAP1 with K = 45 nM and <1 nM, respectively. The ap-
parent discrimination of 49b and 6 for cIAP1 over XIAP was
fascinating and, along lines argued by others, may derive from
minor variations in the P4 binding pockets on BIR domains
5
3,54
within these proteins.
To probe this further we employed
computational techniques. We studied the molecular dynamics
(MD) of 49a/b docked into the binding site of cIAP1 as well
as XIAP (see SI for computational details). We found that 49b
2
buried 50 Å more surface area compared to 49a when aver-
aged over the last 20 ns of a 100 ns MD simulation using
cIAP1-BIR3 as the protein partner (Fig. 5 C & D). This obser-
vation suggested that 49b interacts with cIAP1 more favorably
than 49a and correlated well with competitive binding data.
Comparing MD simulations of ligated cIAP1 and XIAP ex-
plained the observed selectivity for cIAP1. The hydrophobic
binding site in XIAP was unable to accommodate the P4 phe-
nyl substituent, while in cIAP1 it provided a firm anchor for
the ligand: after 75 ns of a 100 ns simulation, the phenyl sub-
stituent exits the XIAP hydrophobic pocket, which then leads
to complex disengagement. Evident from MD data, the hydro-
phobic pocket of cIAP1 can accommodate a larger substituent
relative to the constricted site in XIAP, presumably due to the
steric demand of K206, which corresponds to G306 in cIAP1
EXPERIMENTAL SECTION
Pd(DPEPhos)Cl
pared according to prior literature.
yl)trimethylsilane was prepared according to prior literature. t-
2
was purchased from Strem. Catalyst 16 was pre-
41
(5-bromopent-1-yn-1-
58
Butylhydroperoxide ~5.5 M solution in decane was purchased from
59
Aldrich and iodometrically titrated (c = 5.4 M). Vinyl boronate 22
was prepared using a modified procedure using 1 mol% Schwartz’s
6
0
Reagent. N-hydroxysuccinimide was azeotropically dried from ben-
zene. Fmoc-5-bromo-D-tryptophan and Fmoc-3-methoxy-D-
phenylalanine were synthesized by kinetic enzymatic resolution of
(
see SI Fig. S9 & S10).
6
1
their racemates according to published procedures. L-Phenylalaninol
was purchashed from Chem-Impex. Purification of acidolysis prod-
ucts was performed on an Agilent 1100/1200 HPLC system equipped
with G1361A preparative pumps, a G1314A autosampler, a G1314A
VWD, and a G1364B automated fraction collector. Analytical HPLC
was performed using an identical system, but with a G1312A binary
The method of lead discovery in the above experiments was
highly effective. Using scaffold (+)-5 and an unprotected con-
sensus peptide, we were able to rapidly generate unique mac-
rocyclic ligands for protein surfaces. While this was a proof-
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Page 8 of 20
pump. Mass spectra were recorded using an Agilent 6130 LC/MS
pension was filtered and washed 3x with 1 mL MeOH. The combined
washes were concentrated in vacuo and reconstituted in 1:1
MeOH/CH CN (50 mM in substrate). The clear solution was treated
with piperidine (7 eq.) and Cu(OAc) •H O (7 eq.); the vial was then
capped and heated to 70 ºC. The reaction was monitored by HPLC
and complete within 12 hours. The reaction was concentrated and
purified (see S.I.).
1
2
3
4
5
6
7
8
9
1
1
1
1
1
1
1
1
1
1
2
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2
2
2
2
2
2
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3
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3
3
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3
3
3
3
4
4
4
4
4
4
4
4
4
4
5
5
5
5
5
5
5
5
5
5
6
system equipped with an ESI source. Stationary phase and gradient
profile are noted for individual reactions below. NMR spectra were
recorded on Brüker Avance (300, 400, 500 or 600 MHz) or DRX (500
MHz) spectrometers and calibrated according to the respective resid-
ual solvent peak. 2D-NMR data were acquired as previously
3
2
2
3
4
detailed. High-resolution mass spectra (HRMS) were obtained on a
Thermo Fisher Scientific Exactive Plus (orbitrap) with IonSense ID-
(3-(3-Bromophenyl)prop-1-yn-1-yl)trimethylsilane
CUBE DART, on
a
Q
Exactive™ Plus Hybrid Quadrupole-
In a flame-dried flask under argon, (Trimethylsilyl)acetylene [15.8
mL, 112 mmol] in 80 mL of dry THF was treated with n-BuLi [44.8
mL, 112 mmol, c = 2.5 M] at -78 ºC. While the acetylide solution
stirred, zinc(II) bromide [25.2 g, 112 mmol] was fused under vacuum
then cooled to room temperature under argon. 80 mL of dry THF was
then charged into the flask containing ZnBr . The ZnBr solution was
2 2
then cannulated into the acetylide solution at -78 ºC. The transmeta-
lation was stirred for 30 min then treated with a solution of 3-
bromobenzylbromide [20.0 g, 80 mmol] and Pd(DPEPhos)Cl [115
2
mg, 0.160 mmol] in 80 mL of dry THF – catalyst loading can be in-
creased to drop reaction time and temperature. The solution warmed
to room temperature then heated to 35 ºC. Reaction monitored by H-
NMR. After 2 days, reaction was quenched with saturated aqueous
Orbitrap™ via direct syringe pump injection, or on a Waters LCT
Premier with ACQUITY UPLC (ESI–TOF). Enantiomeric excess of
cyclopropene 17 was assessed using a Mettler Toledo SFC equipped
with a Chiralcel OJ-H column (4.6x250 mm, 5 µm) using 5% i-PrOH
as co-solvent. Flow rate: 2.0 mL/min.
0
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2
3
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9
0
Peptide Synthesis: All peptides were synthesized via either stand-
ard Fmoc solid-phase peptide synthesis using Rink Amide MBHA
resin (polystyrene, 1% DVB, 0.7 mmol/g) or Boc/Cbz solution-phase
3
4
peptide synthesis.
A. Acylation of peptide by template (+)-5 or (+)-S5: A round bot-
tom flask was charged with peptide (1.35 eq.), DMF (1.0 M), and
1
iPr
2
NEt (4.0 – 6.0 eq.), followed by template (1.0 eq.). The reaction
was heated to 40 ºC. Reaction progress was monitored by analytical
HPLC-UV/MS. The reaction was diluted with EtOAc and washed
NH
sat. NH
concentrated in vacuo. A portion of crude was purified by silica
4
Cl and extracted with EtOAc. Organic layer washed twice with
4
Cl, NaHCO , and 1x with brine. Dried with MgSO and
3
4
thrice with NaHCO
with MgSO and concentrated in vacuo.
B. Pictet-Spengler Annulation: Linear precursor was dissolved in
a 4:1 mixture of AcOH/H O (0.2 M) and stirred until HPLC analysis
3
followed by brine. The organic layer was dried
1
4
3
chromatography using hexanes as eluent. H NMR (CDCl , 500
MHz): δ 7.51 (t, J = 7.9 Hz, 1H), 7.38–7.36 (m, 1H), 7.29–7.26 (m,
1H), 7.19 (t, J = 7.9 Hz, 1H), 3.63 (s, 2H), 0.22 (s, 9H); C NMR
13
2
confirmed reaction completion – typically 12 hours. The volatiles
were removed and the residue was concentrated from acetonitrile (3x)
(CDCl
87.8, 25.9, 0.2; HRMS (DART-Orbitrap) m/z: [M-H] calc’d for
14BrSi 265.0054; found 265.0062.
3
, 126 MHz): δ 138.7, 131.1, 130.1, 129.9, 126.6, 122.7, 103.3,
-
followed by CHCl
3
(3x) to remove residual AcOH.
12
C H
C. Friedel-Crafts Macrocyclization: A flask was charged with Pic-
tet-Spengler product (1 eq.) and nitromethane (5 mM in substrate).
The headspace was flushed with argon for 5 mins. TFA (5 vol%) was
then quickly added. Reaction progress was monitored by analytical
HPLC-MS.
1-bromo-3-(propa-1,2-dien-1-yl)benzene
Crude (3-(3-Bromophenyl)prop-1-yn-1-yl)trimethylsilane [32
mmol] was dissolved in 30 mL of DCM and 20 mL of MeOH and
treated with K CO [13.3 g, 96 mmol] under argon. Reaction moni-
2
3
tored by TLC. After 5 hours, the reaction was diluted with DCM and
washed once with water. The aqueous layer was then extracted twice
D. Pd(0)-catalyzed Macrocyclization with Pd(PPh
A flask was charged with Pictet-Spengler product (1 eq.), Cs
eq.), and DMF (5 mM in substrate) and sparged for 30 minutes.
Pd(PPh was then added as a solid, and the solution was sparged for
3
)
4
as catalyst:
2
CO (2
3
with DCM. The combined organic layers were dried with MgSO
The solvent was removed in vacuo. Purified on SiO with hexanes.
1.0 g, 5.2 mmol, 16% yield over two steps (66% isolated yield of 15).
4
.
2
3 4
)
1
another 5 minutes. Reaction progress was monitored by analytical
HPLC-MS. After reaction completion, the reaction was diluted with
H NMR (CDCl
3
, 500 MHz): δ 7.54 (br s, 1H), 7.32 (d, J = 7.8 Hz,
1H), 7.21 (d, J = 7.8 Hz, 1H), 7.17 (t, J = 7.7 Hz, 1H), 6.10 (t, J = 6.6
1
3
EtOAc and washed with 3x NH
was dried with MgSO and concentrated in vacuo.
E. Pd(0)-catalyzed Macrocyclization with [PdCl(C
Xantphos: A flask was charged with Pictet-Spengler product (1 eq.),
Cs CO (2 eq.), and DMF (5 mM in substrate) and sparged for 30
minutes. In a glove bag, a flame-dried Schlenk tube was charged with
PdCl(C )] (9 mg) and Xantphos (37 mg). Outside of the glove-
4
Cl and 1x brine. The organic layer
Hz, 2H), 5.19 (d, J = 6.8 Hz, 1H); C NMR (CDCl
3
, 126 MHz): δ
4
210.1, 136.4, 130.2, 129.9, 129.6, 125.4, 122.9, 93.1, 79.5; HRMS
+
3
H
5
)]
2
/
(DART-Orbitrap) m/z: [M+H] calc’d for C
194.9792.
9 8
H Br 194.9804; found
2
3
1-Bromo-3-(prop-2-yn-1-yl)benzene (15)
Crude (3-(3-Bromophenyl)prop-1-yn-1-yl)trimethylsilane [80
mmol] was dissolved in 400 mL of dry THF and treated with acetic
acid [9.2 mL, 160 mmol] followed by slow addition of tetrabu-
tylammonium fluoride [160 mL, 160 mmol] under argon. Reaction
[
3
H
5
2
bag, the Schlenk tube was charged with 9 mL of 1:1 THF/DMF,
which had been separately sparged for 1 hour. The catalyst solution
was stirred for 5 minutes under Ar and 4 mol% Pd was added to the
reaction flask via syringe. Reaction progress was monitored by ana-
lytical HPLC-MS. After reaction completion, the reaction was diluted
1
monitored by H-NMR. After 15–30 min, THF was removed in vac-
uo and the residue reconstituted in diethyl ether. Organic layer
washed twice with water, sat. NaHCO , and 1x with brine. Ether was
3
completely removed in vacuo, the orange oil was dissolved in pen-
tane, and passed through a plug of silica to remove residual tetrabu-
tylammonium salts. Pentane was removed in vacuo and the colorless
with EtOAc and washed with 3x NH
4
Cl and 1x brine. The organic
layer was dried with MgSO and concentrated in vacuo.
4
F. Copper(I)-Catalyzed Huisgen Cycloaddition:
A vial was charged with macrocyclic compounds (1 eq.), azidoglu-
copyranoside (1.5 eq.), and DMF (0.03 M). The solution was sparged
for 10 minutes. In a separate vial, a stock solution of copper was pre-
pared. Copper iodide was added to a vial and evacuated and back-
filled with argon (3x). DMF (2 mL) was added and the suspension
residue was distilled between 67–69 ºC at 4.7 torr. 12.0 g, 85% yield
1
over two steps. H NMR (CDCl
3
, 500 MHz): δ 7.54 (br s, 1H), 7.40–
7.38 (m, 1H), 7.30–7.27 (m, 1H), 7.20 (t, J = 7.8 Hz, 1H), 3.59 (dd, J
1
3
= 2.7, 0.5 Hz, 2H), 2.25 (t, J = 2.8 Hz, 1H); C NMR (CDCl
3
, 126
MHz): δ 138.4, 131.0, 130.1, 1230.0, 126.6, 122.7, 81.1, 71.3, 24.5;
+
was sparged for 5 minutes. Et
3
N (1 mL) was added to the copper
9 8
HRMS (DART-Orbitrap) m/z: [M+H] calc’d for C H Br 194.9804;
suspension and mixed under sparge for 2 minutes until a homogene-
ous solution was achieved. The copper solution (10 mol% copper)
was then added to the reaction flask. Reaction progress was monitored
by analytical HPLC-MS. After reaction completion, the reaction was
transferred to an HPLC vial. Desired product was isolated by semi-
preparative HPLC purification – see details per example, in S.I.
G. Dimerization of monovalent Smac-mimetics: The TFA-salt of
macrocyclic monomer (1 eq.) was dissolved in 1 mL MeOH and
treated with silica-bound carbonate (2 eq.) for 10 minutes. The sus-
found 194.9797.
(+)-Ethyl (S)-2-(3-bromobenzyl)cycloprop-2-ene-1-carboxylate
(17)
A flame dried flask was charged with freshly distilled 15 [13.0 g,
66.6 mmol], catalyst 16 [76 mg, 0.056 mmol), and 320 mL of dry
DCM. A solution of ethyl diazoacetate [2.5 g, 22.2 mmol] in 50 mL
of dry DCM was added over 12 hours under an argon atmosphere via
syringe pump. After addition, DCM was completely removed in
vacuo and residue dissolved in 25 mL hexanes and loaded onto a
ACS Paragon Plus Environment

Page 9 of 20
The Journal of Organic Chemistry
silica column. Column was eluted with hexanes until removal of
starting alkyne. Column then eluted with a gradient from 2% à 6%
EtOAc in hexanes. Product was obtained as a yellow oil [5.0 g, 17.5
Dioxane and deionized water were sparged with argon for one
hour. Crude acetal S18 [16.4 mmol], vinyl boronate 22 [5.9 g, 19.7
mmol], and Na CO [5.2 g, 49.2 mmol] were dissolved in 30 mL of
:1 dioxane/water. The system was sparged for 15 min, charged with
Pd(PPh [190 mg, 0.164 mmol], and sparged an additional 15 min.
The system was then taken to reflux and monitored by H-NMR.
After two days, the reaction was complete. Dioxane was removed in
vacuo, exchanged for EtOAc, and this solution was washed 3x with
1
2
3
4
5
6
7
8
9
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
2
2
2
3
3
3
3
3
3
3
3
3
3
4
4
4
4
4
4
4
4
4
4
5
5
5
5
5
5
5
5
5
5
6
2
3
!
"
1
5
mmol]. 80% yield, 95% e.e. [α] = +22.2º, c = 4.26, CHCl
3
, H
, 500 MHz): δ 7.37 (br s, 1H), 7.32 (d, J = 7.3 Hz, 1H),
.16–7.11 (m, 2H), 6.47 (br s, 1H), 4.05 (q, J = 7.2 Hz, 2H), 3.79 &
.72 (AB quartet, J = 17.6 Hz, 2H), 2.19 (br s, 1H), 1.17 (t, J = 7.2
!
3 4
)
NMR (CDCl
3
1
7
3
1
3
Hz, 3H); C NMR (CDCl
3
, 126 MHz): δ 176.0, 138.6, 131.8, 130.3,
water and 1x with brine. The organic layer was dried with MgSO
4
130.1, 127.4, 122.7, 114.1, 96.5, 60.5, 31.1, 20.5, 14.5; HRMS
+
and concentrated to dryness. The crude product was dissolved in
hexanes and chromatographed using a gradient of 0% à 5% EtOAc
in hexanes. Collected 23 [2.85 g, 4.73 mmol] as a colorless oil. 29%
(
DART-Orbitrap) m/z: [M+H] calc’d for C13
found 281.0163.
5-(Trimethylsilyl)pent-4-yn-1-yl)magnesium bromide (18)
14 2
H O Br 281.0172;
(
!
"
1
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
yield from 17. [α] = +3.05º, c = 0.46, CHCl , H NMR (CDCl , 500
3
3
A flask equipped with a condenser was charged with magnesium
[2.24 g, 85.4 mmol] then flame-dried under vacuum then flushed with
argon. After cooling, the flask was charged with 14 mL of dry THF
and treated with an iodine crystal. The flask was then heated with a
heatgun until complete dissolution of the orange iodine color. Neat
!
MHz): δ 7.22–7.18 (m, 3H), 7.09 (ddd, J = 7.2, 1.4, 1.4 Hz, 1H), 6.55
(ddd, J = 15.9, 1.5, 1.5 Hz, 1H), 6.25 (ddd, J = 15.9, 5.0, 5.0 Hz, 1H),
4.34 (dd, J = 5.0, 1.7 Hz, 2H), 4.28 (s, 1H), 4.10 (q, J = 7.1 Hz, 2H),
3.82–3.74 (m, 2H), 3.51–3.41 (m, 2H), 2.91 & 2.82 (AB quartet, J =
13.4 Hz, 2H), 2.32 (s, 2H), 2.16 (dd, J = 6.7, 6.7 Hz, 2H), 1.71–1.48
(m, 6H), 1.28–1.19 (m, 11H), 0.94 (s, 9H), 0.13 (s, 9H), 0.11 (s, 6H);
(
5-bromopent-1-yn-1-yl)trimethylsilane [9.36 g, 42.7 mmol] was then
added at a rate sufficient to maintain a slight reflux. After complete
addition, the system was heated to reflux for an additional 30 min.
The Grignard reagent was measured to be 1.44 M by titration with
1
3
3
C NMR (CDCl , 126 MHz): δ 172.7, 138.7, 136.7, 130.1, 129.8,
129.3, 128.9, 128.1, 124.2, 108.2, 107.8, 84.4, 66.3, 66.2, 64.0, 60.1,
45.6, 39.6, 37.5, 33.3, 26.2, 26.1, 24.9, 23.7, 20.9, 18.6, 15.7, 15.7,
6
2
menthol/phenanthroline.
+
Ethyl (S)-3-(3-bromobenzyl)-3-formyl-8-(trimethylsilyl)oct-7-
ynoate (21)
14.4, 0.3, -5.0; HRMS (QE Orbitrap) m/z: [M-tert-Bu] calc’d for
C
30
H
49
O
5
Si
2
545.3113; found 545.3115.
A flame-dried flask was charged with solid copper(I) iodide [6.25
g, 32.8 mmol] then evacuated and backfilled with argon 3x. The flask
was charged with 131 mL of dry THF and TMEDA [5.4 mL, 36.1
mmol] and stirred at room temperature for 30 minutes then cooled to -
Ethyl
(S,E)-3-(diethoxymethyl)-3-(3-(3-hydroxyprop-1-en-1-
yl)benzyl)oct-7-ynoate (S19)
Pure 23 [2.29 g, 3.80 mmol] was dissolved in 40 mL of dry THF
and cooled to 0 ºC. A solution of TBAF [9.5 mL, 9.50 mmol] was
slowly added over 5 minutes. The reaction was monitored by TLC.
After 30 minutes, THF was removed in vacuo and exchanged for
EtOAc, and this solution was washed 3x with water and 1x with
brine. Organic layer was dried with MgSO and concentrated in vac-
4
uo to provide S19 as a yellow oil, which was carried forward without
purification.
(S,E)-3-(Diethoxymethyl)-3-(3-(3-hydroxyprop-1-en-1-
yl)benzyl)oct-7-ynoic acid (24)
Crude ethyl (S,E)-3-(diethoxymethyl)-3-(3-(3-hydroxyprop-1-en-1-
yl)benzyl)oct-7-ynoate [3.80 mmol] was dissolved in 38 mL of 2:1
2
EtOH/H O and treated with KOH [2.13 g, 38.0 mmol]. The ensuing
red solution was then heated to 50 ºC overnight. After stirring for 12
hours, the reaction was complete. Solvent was removed and the red
4
5 ºC. Previously prepared Grignard reagent (18) [32.8 mmol] was
added to the reaction flask and stirred an additional 30 minutes at -45
ºC. Cyclopropene (+)-17 [4.6 g, 16.4 mmol] in 33 mL of dry THF
was added to the reaction flask at -45 ºC and stirred for 30 minutes. In
a separate flame-dried flask, t-butylhydroperoxide [6 mL, 32.8 mmol,
c = 5.4 M] was dissolved in 82 mL of dry THF, cooled to -78 ºC, and
treated with n-BuLi [13.4 mL, 33.6 mmol, c = 2.5 M]. After complete
carbometalation as determined by TLC, the reaction flask was cooled
to -78 ºC and treated with previously prepared t-BuOOLi via cannula-
tion. The reaction was stirred at this temperature for one hour – sig-
1
nificant decomposition was observed by H-NMR at longer time
points – then cannulated into a cold solution of 2:1 NH
and extracted with EtOAc. The organic layer was washed 3x with
water, 1x with brine, dried with MgSO , and concentrated in vacuo to
4
Cl / NH
4
OH
4
oil was treated with 200 mL of 0.3 N NaH
EtOAc. The combined organic layers were washed with brine, dried
with MgSO and concentrated in vacuo. The red oil was carried for-
2 4
PO and extracted 3x with
give 21 as a green oil, which was carried forward without purification.
Ethyl (S)-2-(3-bromobenzyl)cycloprop-2-ene-1-carboxylate (S1)
A 1 mL aliquot of the above carbometalation was removed and
4
ward without purification.
quenched with 2:1 NH
4
Cl/NH
4
OH and worked up as above. pTLC to
(+)-2,5-Dioxopyrrolidin-1-yl (S,E)-3-(diethoxymethyl)-3-(3-(3-
((isobutoxycarbonyl)oxy)prop-1-en-1-yl)benzyl)oct-7-ynoate (5)
Crude cinnamyl alcohol 24 [3.80 mmol] was dissolved in 7.6 mL
of dry DCM, treated with N-methylmorpholine [1.88 mL, 17.1
mmol], and cooled to -5 ºC under argon. i-Butyl chloroformate [1.04
mL, 7.98 mmol] was then added over two minutes. The reaction was
monitored by TLC for full conversion to the di-carbonate species. At
this time, solid N-hydroxysuccinimide [875 mg, 7.60 mmol] was
added to the reaction flask. The ice in the cold bath was replenished
and the reaction was allowed to slowly warm overnight. Twelve
hours after addition of NHS, solid DMAP [1.39 g, 11.4 mmol] was
added to decompose byproduct, i-butyl succinimidyl carbonate. After
stirring with DMAP for 10 min, the reaction was quenched with Na-
remove Grignard byproducts: 4% EtOAc in hexanes. Product was
1
obtained as a colorless residue. H NMR (CDCl
7
2
3
, 500 MHz): δ 7.33–
.30 (m, 2H), 7.15–7.11 (m, 2H), 4.17 (q, J = 7.1 Hz, 2H), 2.90 &
.84 (AB quartet, J = 15.3 Hz, 2H), 2.16 (t, J = 7.3 Hz, 2H), 1.66 (dd,
J = 8.1, 5.7 Hz, 1H), 1.63–1.57 (m, 2H), 1.36–1.33 (m, 1H), 1.32–
1.30 (m, 1H), 1.29 (t, J = 7.2 Hz, 3H), 1.28–1.25 (m, 1H), 1.00 (dd, J
1
3
=
8.1, 4.6 Hz, 1H); C NMR (CDCl
3
, 126 MHz): δ 176.0, 138.6,
1
31.8, 130.3, 130.1, 127.4, 122.7, 114.1, 96.5, 60.5, 31.1, 20.5, 14.5;
+
HRMS (ESI-QE Orbitrap) m/z: [M+H] calc’d for C21
21.1193; found 421.1190.
Ethyl (S)-3-(3-bromobenzyl)-3-(diethoxymethyl)-8-
2
H30BrO Si
4
(trimethylsilyl)oct-7-ynoate
Crude aldehyde ethyl (S)-2-(3-bromobenzyl)cycloprop-2-ene-1-
carboxylate [16.4 mmol] was dissolved in 50 mL of dry ethanol then
treated with ethyl orthoformate [8.2 mL, 49.2 mmol] and p-TSA [312
mg, 1.64 mmol]. The reaction was heated to 60 ºC and monitored by
HCO
NaHCO
vacuo. The crude residue was dissolved in a minimum amount of 3:1
hexanes/CHCl and loaded onto silica column. Elution with a gradi-
3
and extracted with EtOAc. The organic layer washed 2x with
3
and 1x with brine, dried with MgSO , and concentrated in
4
3
1
H-NMR. The reaction was complete within 1 hour. Ethanol was
ent of 5% → 30% EtOAc/hexanes provided (+)-5 [1.15 g, 1.96 mmol]
as a colorless oil. 52% from 23, 94% e.e. as determined by diastere-
removed in vacuo and the residue reconstituted in EtOAc then washed
!
"
1
3
x with NaHCO
3
and 1x with brine. The organic layer was dried with
omeric derivatization – see SI. [α] = +8.56º, c = 0.58, CHCl
3
, H
, 500 MHz): δ 7.28–7.21 (m, 3H), 7.15 (d, J = 7.3 Hz,
H), 6.68 (d, J = 15.9 Hz, 1H), 6.30 (ddd, J = 15.9, 6.4, 6.4 Hz, 1H),
!
MgSO and concentrated in vacuo to give a yellow oil. Acetal S18
4
NMR (CDCl
3
was carried forward without purification.
Ethyl (S,E)-3-(3-(3-((tert-butyldimethylsilyl)oxy)prop-1-en-1-
yl)benzyl)-3-(diethoxymethyl)-8-(trimethylsilyl)oct-7-ynoate (23)
1
4
3
.77 (d, J = 6.4 Hz, 2H), 4.34 (s, 1H), 3.93 (d, J = 6.7 Hz, 2H), 3.83–
.78 (m, 2H), 3.54–3.44 (m, 2H), 2.92 & 2.86 (AB quartet, J = 14.3
Hz, 2H), 2.84 (br s, 4H), 2.63 (s, 2H), 2.14 (ddd, J = 6.5, 6.5, 2.4 Hz,
ACS Paragon Plus Environment

The Journal of Organic Chemistry
Page 10 of 20
2
H), 2.00–1.95 (m, 1H), 1.76–1.55 (m, 5H), 1.25–1.19 (m, 6H), 0.95
washed 3x with water and 1x with brine. Organic layer was dried
with MgSO and concentrated in vacuo to provide ethyl (S,E)-4,4-
1
3
1
2
3
4
5
6
7
8
9
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
2
2
2
3
3
3
3
3
3
3
3
3
3
4
4
4
4
4
4
4
4
4
4
5
5
5
5
5
5
5
5
5
5
6
(d, J = 6.7 Hz, 6H); C NMR (CDCl
3
, 126 MHz): δ 169.3, 167.7,
4
1
8
2
55.4, 138.1, 136.0, 135.0, 130.9, 129.5, 128.4, 124.8, 122.6, 107.7,
4.6, 74.3, 68.5, 68.5, 66.6, 66.3, 45.6, 39.3, 34.2, 33.1, 27.9, 25.7,
diethoxy-3-(3-(3-hydroxyprop-1-en-1-yl)benzyl)-3-methylbutanoate
as a yellow oil, which was carried forward without purification.
(S,E)-4,4-Diethoxy-3-(3-(3-hydroxyprop-1-en-1-yl)benzyl)-3-
methylbutanoic acid (S4)
3.54, 19.3, 19.0, 15.6, 15.6; MS m/z HRMS (DART-Orbitrap) m/z:
-
[
M-H] calc’d for C32
H
42NO
9
584.2865; found 584.2871.
Ethyl (S)-3-(3-bromobenzyl)-3-methyl-4-oxobutanoate (S2)
A flame-dried flask was charged with solid copper(I) iodide [6.8 g,
5.7 mmol] then evacuated and backfilled with argon 3x. The flask
Crude
yl)benzyl)-3-methylbutanoate [6.27 mmol] was dissolved in 63 mL of
2:1 EtOH/H O and treated with KOH [3.5 g, 62.7 mmol]. The ensu-
ethyl
(S,E)-4,4-diethoxy-3-(3-(3-hydroxyprop-1-en-1-
3
2
was charged with 179 mL of dry THF and TMEDA [5.9 mL, 39.3
mmol] and stirred at room temperature for 30 minutes then cooled to -
45 ºC. Methylmagnesium bromide [14.9 mL, 35.7 mmol] was added
to the reaction flask and stirred an additional 30 minutes at -45 ºC.
Cyclopropene (+)-17 [5.0 g, 17.9 mmol] in 36 mL of dry THF was
added to the reaction flask at -45 ºC and stirred for 30 minutes. In a
separate flame-dried flask, t-butylhydroperoxide [7.9 mL, 42.8 mmol,
c = 5.4 M] was dissolved in 107 mL of dry THF, cooled to -78 ºC,
and treated with n-BuLi [21 mL, 44.6 mmol, c = 2.5 M]. After com-
plete carbometalation as determined by TLC, the reaction flask was
cooled to -78 ºC and treated with previously prepared t-BuOOLi via
cannulation. The reaction was stirred at this temperature for one hour
ing red solution was then heated to 50 ºC overnight. After stirring for
12 hours, the reaction was complete. Solvent was removed and the
red oil was treated with 200 mL of 0.3 N NaH
with EtOAc. The combined organic layers were washed with brine,
dried with MgSO and concentrated in vacuo. The red oil was carried
2 4
PO and extracted 3x
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
4
forward without purification.
(+)-2,5-Dioxopyrrolidin-1-yl
(S,E)-4,4-diethoxy-3-(3-(3-
((isobutoxycarbonyl)oxy)prop-1-en-1-yl)benzyl)-3-
methylbutanoate (S5)
Crude (S,E)-4,4-Diethoxy-3-(3-(3-hydroxyprop-1-en-1-yl)benzyl)-
3-methylbutanoic acid [1.58 mmol] was dissolved in 3.2 mL of dry
DCM, treated with N-methylmorpholine [782 µL, 7.11 mmol], and
cooled to -5 ºC under argon. i-Butyl chloroformate [431 µL, 3.32
mmol] was then added. The reaction was monitored by TLC for full
conversion to the di-carbonate species. At this time, solid N-
hydroxysuccinimide [364 mg, 3.16 mmol] was added to the reaction
flask. The ice in the cold bath was replenished and the reaction was
allowed to slowly warm overnight. Twelve hours after addition of
NHS, solid DMAP [579 mg, 4.74 mmol] was added to decompose by-
product, i-butyl succinimidyl carbonate. After stirring with DMAP for
–
significant decomposition was observed at longer reaction times –
then cannulated into a cold solution of 2:1 NH Cl / NH OH and ex-
tracted with EtOAc. The organic layer was washed 3x with water, 1x
with brine, dried with MgSO , and concentrated in vacuo to give a
4
4
4
green oil, which was carried forward without purification.
Ethyl (S)-3-(3-bromobenzyl)-4,4-diethoxy-3-methylbutanoate
Crude ethyl (S)-3-(3-bromobenzyl)-3-methyl-4-oxobutanoate [17.9
mmol] was dissolved in 89 mL of dry ethanol then treated with ethyl
orthoformate [8.9 mL, 53.6 mmol] and p-TSA [340 mg, 1.79 mmol].
The reaction was heated to 60 ºC and monitored by H-NMR. The
reaction was complete within 1 hour. Ethanol was removed in vacuo,
10 min, reaction quenched with NaHCO
Organic layer washed 2x with NaHCO and 1x with brine, dried with
MgSO , and concentrated in vacuo. The crude residue was dissolved
in a minimum amount of 3:1 hexanes/CHCl and loaded onto silica
column. Elution with a gradient of 5% → 20% EtOAc/hexanes pro-
vided (+)-2,5-Dioxopyrrolidin-1-yl (S,E)-4,4-diethoxy-3-(3-(3-
3
and extracted with EtOAc.
1
3
4
and the residue was reconstituted in EtOAc then washed 3x with Na-
3
HCO
and concentrated in vacuo to give a yellow oil. Acetal product [6.4 g,
6.4 mmol, 92% crude recovery] was carried forward without purifi-
cation.
Ethyl (S,E)-3-(3-(3-((tert-butyldimethylsilyl)oxy)prop-1-en-1-
3 4
and 1x with brine. The organic layer was dried with MgSO
1
((isobutoxycarbonyl)oxy)prop-1-en-1-yl)benzyl)-3-methylbutanoate
[615 mg, 1.15 mmol] as a white gum. 65% from ethyl (S,E)-3-(3-(3-
((tert-butyldimethylsilyl)oxy)prop-1-en-1-yl)benzyl)-4,4-diethoxy-3-
methylbutanoate, 89% e.e. as determined by diastereomeric derivati-
yl)benzyl)-4,4-diethoxy-3-methylbutanoate (S3)
!
"
1
Dioxane and deionized water were sparged with argon for one
zation – see SI. [α] = +15.71º, c = 0.56, CHCl
3
, H NMR (CDCl
500 MHz): δ 7.28–7.21 (m, 3H), 7.15 (d, J = 7.3 Hz, 1H), 6.68 (d, J =
5.9 Hz, 1H), 6.30 (ddd, J = 15.9, 6.4, 6.4 Hz, 1H), 4.77 (d, J = 6.4
Hz, 2H), 4.34 (s, 1H), 3.93 (d, J = 6.7 Hz, 2H), 3.83–3.78 (m, 2H),
.54–3.44 (m, 2H), 2.92 & 2.86 (AB quartet, J = 14.3 Hz, 2H), 2.84
3
,
!
hour.
methylbutanoate [16.4 mmol], vinyl boronate 22 [5.9 g, 19.7 mmol],
and Na CO [5.2 g, 49.2 mmol] were dissolved in 41 mL of 5:1 diox-
ane/water. The system was sparged for 15 min, charged with
Pd(PPh [190 mg, 0.164 mmol], and sparged an additional 15 min.
The system was then taken to reflux and monitored by H-NMR.
After two days, reaction was complete. Dioxane was removed in
vacuo, exchanged for EtOAc, and washed 3x with water and 1x with
brine. The organic layer was dried with MgSO and concentrated to
4
dryness. The crude product was dissolved in hexanes and chromato-
graphed using a gradient of 0% → 5% EtOAc in hexanes. Collected
product [2.28 g, 4.76 mmol] as a colorless oil. 29% yield from (+)-
Crude
ethyl
(S)-3-(3-bromobenzyl)-4,4-diethoxy-3-
1
2
3
3
3 4
)
1
(br s, 4H), 2.63 (s, 2H), 2.14 (ddd, J = 6.5, 6.5, 2.4 Hz, 2H), 2.00–1.95
(m, 1H), 1.76–1.55 (m, 5H), 1.25–1.19 (m, 6H), 0.95 (d, J = 6.7 Hz,
13
3
6H); C NMR (CDCl , 101 MHz): δ 169.2, 167.6, 155.3, 137.9,
1
6
35.9, 134.9, 130.8, 129.3, 128.3, 124.7, 122.5, 107.3, 74.2, 68.3,
6.7, 65.9, 43.1, 40.1, 36.2, 27.8, 25.6, 19.4, 18.9, 15.51, 15.48.
-
HRMS (DART-Orbitrap) m/z: [M-H] calc’d for C28
H
38NO
9
532.2552;
found 532.2599.
1
(E)-3-(3-((S)-2-(diethoxymethyl)-2-(2-oxo-2-(((R)-1-
phenylethyl)amino)ethyl)hept-6-yn-1-yl)phenyl)allyl
carbonate (S6)
1
7
7. H NMR (CDCl
3
, 500 MHz): δ 7.24–7.18 (m, 3H), 7.05 (ddd, J =
isobutyl
.2, 1.6, 1.6 Hz, 1H), 6.56 (ddd, J = 15.7, 1.9, 1.9 Hz, 1H), 6.26 (ddd,
J = 15.8, 5.1, 5.1 Hz, 1H), 4.35 (dd, J = 5.1, 1.8 Hz, 2H), 4.30 (s, 1H),
4.13 (q, J = 7.2 Hz, 2H), 3.85–3.78 (m, 2H), 3.55–3.45 (m, 2H), 2.86
&
1
6
1
40.7, 39.4, 26.1, 19.8, 18.6, 15.7, 15.6, 14.4, -5.0. HRMS (QE Or-
bitrap) m/z: [M+Na] calc’d for C27
5
Synthesized according to General Procedure A using (R)-(+)-
phenylethylamine and (+)-5 [77 µmol]. Obtained 20 mg [crude, 45%]
2.82 (AB quartet, J = 13.2 Hz, 2H), 2.35 & 2.28 (AB quartet, J =
of
(E)-3-(3-((S)-2-(diethoxymethyl)-2-(2-oxo-2-(((R)-1-
4.9 Hz, 2H) 1.28–1.22 (m, 9H), 1.00 (s, 3H), 0.94 (s, 9H), 0.11 (s,
13
phenylethyl)amino)ethyl)hept-6-yn-1-yl)phenyl)allyl isobutyl car-
H); C NMR (CDCl
3
, 126 MHz): δ 172.9, 138.6, 136.8, 130.2,
1
bonate. The crude was then dissolved in 500 µL of CDCl
3
for H-
NMR analysis of diastereomeric mixture. H NMR (CDCl , 500
MHz): δ 7.34–7.31 (m, 4H), 7.28 (dd, J = 1.8, 1.8 Hz, 1H), 7.25–7.23
m, 2H), 7.20 (dd, J = 7.5, 7.5 Hz, 1H), 7.16 (ddd, J = 7.4, 1.5, 1.5
Hz, 1H), 6.65 (ddd, J = 15.9, 1.2, 1.2 Hz, 1H), 6.34 (d, J = 7.3 Hz,
29.8, 129.2, 129.0, 128.1, 124.2, 108.2, 66.7, 66.0, 64.1, 60.1, 42.9,
1
3
+
H
46
O
5
SiNa 501.3007; found
(
01.2991.
Ethyl
(S,E)-4,4-diethoxy-3-(3-(3-hydroxyprop-1-en-1-
1
1
2
9
H), 6.28 (ddd, J = 15.9, 6.4, 6.4 Hz, 1H), 5.07 (quint., J = 7.0 Hz,
H), 4.77 (dd, J = 6.5, 1.3 Hz, 2H), 4.16 (s, 1H), 3.94 (d, J = 6.6 Hz,
H), 3.73–3.65 (m, 2H), 3.47 (dd, J = 9.2, 7.0 Hz, 1H), 3.30 (dd, J =
.0, 7.0 Hz, 1H), 2.85 & 2.72 (AB quartet, J = 13.4 Hz, 2H), 2.31 &
yl)benzyl)-3-methylbutanoate
Pure ethyl (S,E)-3-(3-(3-((tert-butyldimethylsilyl)oxy)prop-1-en-1-
yl)benzyl)-4,4-diethoxy-3-methylbutanoate [3.0 g, 6.27 mmol] was
dissolved in 21 mL of dry THF and cooled to 0 ºC. A solution of
TBAF [14.0 mL, 13.8 mmol] was slowly added over 5 minutes. The
reaction was monitored by TLC. After 30 minutes, THF was re-
moved in vacuo and exchanged for EtOAc, and the solution was
2.13 (AB quartet, J = 14.0 Hz, 2H), 2.18–2.13 (m, 1H), 2.01–1.95 (m,
1
1
H), 1.95 (t, J = 2.7 Hz, 1H), 1.83–1.70 (m, 2H), 1.62–1.49 (m, 2H),
.48 (d, J = 6.9 Hz, 1H), 1.15 (t, J = 7.0 Hz, 3H), 1.14 (t, J = 6.9 Hz,
ACS Paragon Plus Environment

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The Journal of Organic Chemistry
1
3
3
H), 0.95 (d, J = 6.7 Hz, 6H); C NMR (CDCl
3
, 126 MHz): δ 171.2,
(E)-3-(3-((S)-2-(2-(((S)-1-amino-3-(1H-indol-3-yl)-1-oxopropan-
1
2
3
4
5
6
7
8
9
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
2
2
2
3
3
3
3
3
3
3
3
3
3
4
4
4
4
4
4
4
4
4
4
5
5
5
5
5
5
5
5
5
5
6
155.3, 143.8, 138.6, 135.7, 135.1, 131.0, 129.6, 128.7, 128.2, 127.3,
2-yl)amino)-2-oxoethyl)-2-(diethoxymethyl)hept-6-yn-1-
yl)phenyl)allyl isobutyl carbonate (26)
Synthesized according to General Procedure A using L-tryptophan
amide and (+)-5 [51 µmol]. Obtained 30 mg [45 µmol, 87% crude
recovery] of 26.
(E)-3-(3-(((1S,5S,11bR)-5-carbamoyl-3-oxo-1-(pent-4-yn-1-yl)-
2,3,5,6,11,11b-hexahydro-1H-indolizino[8,7-b]indol-1-
yl)methyl)phenyl)allyl isobutyl carbonate (27)
Synthesized according to General Procedure B using crude material
from the previous reaction. After reaction completion, solvent was
removed, and the crude residue was dissolved in ~500 µL DMSO and
1
4
26.4, 124.5, 122.4, 108.6, 84.6, 74.3, 68.6, 68.4, 67.2, 65.5, 48.8,
5.9, 40.3, 39.6, 32.7, 27.9, 23.2, 21.9, 19.2, 19.0, 15.7, 15.6; MS m/z:
+
[
M+Na] calc’d for C36
H
49NO
6
Na 614.3; found 614.4.
(
E)-3-(3-((S)-2-(diethoxymethyl)-2-(2-oxo-2-(((S)-1-
phenylethyl)amino)ethyl)hept-6-yn-1-yl)phenyl)allyl
carbonate (S7)
isobutyl
Synthesized according to General Procedure A using (S)-(-)-
phenylethylamine and (+)-5 [77 µmol]. Obtained 22 mg [crude, 49%]
of
(E)-3-(3-((S)-2-(diethoxymethyl)-2-(2-oxo-2-(((S)-1-
phenylethyl)amino)ethyl)hept-6-yn-1-yl)phenyl)allyl isobutyl car-
1
bonate. The crude was then dissolved in 500 µL of CDCl
3
for H-
NMR analysis of diastereomeric mixture. H NMR (CDCl , 500
MHz): δ 7.35–7.32 (m, 4H), 7.28 (dd, J = 1.7, 1.7 Hz, 1H), 7.27–7.22
m, 2H), 7.20–7.17 (m, 2H), 6.64 (ddd, J = 15.9, 1.2, 1.2 Hz, 1H),
purified by semi-preparative HPLC – see SI for details – to give 12
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
1
3
mg [21 µmol, 51% yield] of desired 27. H NMR (DMSO-d
6
, 600
MHz): δ 10.86 (s, 1H), 7.48 (s, 1H), 7.41 (d, J = 6.6 Hz, 1H), 7.41 (d,
J = 6.6 Hz, 1H), 7.23 (d, J = 6.4 Hz, 1H), 7.14–7.11 (m, 1H), 7.12–
7.09 (m, 1H), 7.09 (s, 1H), 7.00 (dd, J = 6.3, 6.3 Hz, 1H), 6.80 (d, J =
5.9 Hz, 1H), 6.71 (s, 1H), 6.46 (d, J = 15.5 Hz, 1H), 6.13–6.10 (m,
1H), 5.06 (s, 1H), 4.82 (d, J = 4.5 Hz, 1H), 4.68 (d, J = 4.1 Hz, 2H),
3.90 (d, J = 4.5 Hz, 2H), 3.35 (d, J = 15.0 Hz, 1H), 2.78 (s, 1H), 2.65
& 2.17 (AB quartet, J = 15.9 Hz, 2H), 2.55–2.52 (m, 1H), 2.33–2.31
(m, 1H), 2.33–2.31 (m, 1H), 2.24–2.20 (m, 1H), 2.21–2.19 (m, 1H),
1.95–1.90 (m, 1H), 1.92–1.89 (m, 1H), 1.88–1.83 (m, 1H), 1.74–1.70
(
6.38 (d, J = 7.4 Hz, 1H), 6.27 (ddd, J = 15.9, 6.5, 6.5 Hz, 1H), 5.07
(quint., J = 7.0 Hz, 1H), 4.76 (dd, J = 6.5, 1.3 Hz, 2H), 4.17 (s, 1H),
3
9
.93 (d, J = 6.7 Hz, 2H), 3.74 (dd, J = 8.9, 6.9 Hz, 1H), 3.66 (dd, J =
.2, 6.9 Hz, 1H), 3.42 (dd, J = 9.0, 7.0 Hz, 1H), 3.38 (dd, J = 9.1, 7.0
Hz, 1H), 2.87 & 2.74 (AB quartet, J = 13.4 Hz, 2H), 2.29 & 2.16 (AB
quartet, J = 14.1 Hz, 2H), 2.10–2.02 (m, 1H), 2.01–1.94 (m, 1H), 1.95
(t, J = 2.6 Hz, 1H), 1.78–1.69 (m, 2H), 1.61–1.45 (m, 3H), 1.47 (d, J
1
3
=
6.9 Hz, 1H), 1.22 (t, J = 7.0 Hz, 3H), 1.01 (t, J = 7.0 Hz, 3H), 0.95
(m, 1H), 1.60–1.56 (m, 1H), 0.9 (d, J = 4.9 Hz, 6H); C NMR
(DMSO-d , 151 MHz): δ 171.9, 171.6, 154.4, 137.2, 136.6, 135.0,
1
3
(
d, J = 6.7 Hz, 6H); C NMR (CDCl
3
, 126 MHz): δ 171.2, 155.3,
43.8, 138.6, 135.7, 135.1, 131.1, 129.6, 128.7, 128.2, 127.3, 126.4,
24.5, 122.4, 108.6, 84.6, 74.3, 68.6, 68.4, 66.9, 65.8, 48.9, 45.9,
6
1
1
133.4, 130.0, 128.8, 128.5, 127.6, 125.8, 124.2, 122.8, 120.9, 118.3,
117.5, 111.1, 106.9, 84.2, 73.0, 71.1, 67.5, 59.2, 49.3, 45.3, 39.9,
40.3, 39.5, 33.0, 27.9, 23.2, 21.9, 19.2, 19.0, 15.7, 15.6; MS m/z:
38.6, 34.5, 26.9, 23.8, 22.9, 18.2, 18.0; HRMS (QE Orbitrap) m/z:
+
+
[M+Na] calc’d for C36
H
49NO
6
Na 614.3; found 614.4.
[M+Na] calc’d for C35
H
39
N
3
O
5
Na 604.2782; found 604.2783.
(
E)-3-(3-((S)-2-(diethoxymethyl)-2-methyl-4-oxo-4-(((R)-1-
(S)-5-(((S)-1-amino-3-(4-hydroxyphenyl)-1-oxopropan-2-
yl)amino)-4-((S)-2-((S)-4,4-diethoxy-3-(3-((E)-3-
((isobutoxycarbonyl)oxy)prop-1-en-1-yl)benzyl)-3-
methylbutanamido)-3-(1H-indol-3-yl)propanamido)-5-
oxopentanoic acid (28)
Synthesized according to Procedure A beginning with 0.71 mmol
of (+)-S5. Carried forward without purification.
(S)-5-(((S)-1-amino-3-(4-hydroxyphenyl)-1-oxopropan-2-
yl)amino)-4-((1S,5S,11bR)-1-(3-((E)-3-
phenylethyl)amino)butyl)phenyl)allyl isobutyl carbonate (S8)
Synthesized according to General Procedure A using (R)-(+)-
phenylethylamine and (+)-S5 [47 µmol]. Obtained 22 mg [crude,
87%] of (E)-3-(3-((S)-2-(diethoxymethyl)-2-methyl-4-oxo-4-(((R)-1-
phenylethyl)amino)butyl)phenyl)allyl isobutyl carbonate. The crude
1
was then dissolved in 500 µL of CDCl
3
for H-NMR analysis of dia-
1
stereomeric mixture. H NMR (CDCl
H), 7.28–7.20 (m, 4H), 7.14 (ddd, J = 6.9, 1.5, 1.5 Hz, 1H), 6.66 (d,
J = 15.9 Hz, 1H), 6.28 (ddd, J = 15.9, 6.5, 6.5 Hz, 1H), 6.03 (d, J =
3
, 500 MHz): δ 7.34–7.32 (m,
4
((isobutoxycarbonyl)oxy)prop-1-en-1-yl)benzyl)-1-methyl-3-oxo-
2,3,5,6,11,11b-hexahydro-1H-indolizino[8,7-b]indole-5-
carboxamido)-5-oxopentanoic acid (29)
7
2
3
.8 Hz, 1H), 5.12 (quint., J = 7.2 Hz, 1H), 4.77 (dd, J = 6.5, 1.1 Hz,
H), 4.22 (s, 1H), 3.94 (d, J = 6.2 Hz, 2H), 3.77–3.70 (m, 2H), 3.52–
.49 (m, 1H), 3.32–3.27 (m, 1H), 2.86 & 2.74 (AB quartet, J = 13.0
Synthesized according to Procedure B beginning with 58 mg of
Hz, 2H), 2.25 & 2.09 (AB quartet, J = 13.9 Hz, 2H), 2.02–1.94 (m,
1H), 1.49 (d, J = 6.9 Hz, 3H), 1.21 (t, J = 7.0 Hz, 3H), 1.15 (t, J = 7.0
crude 28. Chromatographed on SiO
MeOH in CHCl (0.5% AcOH). White Solid. 34 mg, [41 µmol, 94%
yield]. H NMR (DMSO-d , 500 MHz): δ12.07 (br s, 1H), 10.93 (s,
2
with a gradient from 0% to 5%
3
1
3
1
Hz, 3H), 0.96 (d, J = 6.8 Hz, 6H), 0.95 (s, 3H); C NMR (CDCl
3
, 126
6
MHz): δ 171.1, 155.4, 143.6, 138.7, 135.7, 135.2, 131.2, 129.7, 128.7,
1H), 9.15 (s, 1H), 8.19 (d, J = 7.6 Hz, 1H), 7.67 (d, J = 7.8 Hz, 1H),
7.43 (d, J = 7.6 Hz, 1H), 7.37 (d, J = 7.9 Hz, 1H), 7.31 (d, J = 7.9 Hz,
1H), 7.30 (br s, 1H), 7.22 (dd, J = 7.6, 7.6 Hz, 1H), 7.08 (dd, J = 7.6,
7.6 Hz, 1H), 7.01–6.98 (m, 2H), 6.94–6.88 (m, 4 H), 6.63–6.54 (m,
4H), 6.26 (ddd, J = 15.9, 6.3, 6.3 Hz, 1H), 5.09 (d, J = 6.9 Hz, 1H),
5.01 (s, 1H), 4.72, (d, J = 6.1 Hz, 1H), 4.24 (ddd, J = 7.5, 7.5, 7.5 Hz,
1H), 4.08 (ddd, J = 7.9, 7.9, 7.9 Hz, 1H), 3.90 (d, J = 6.7 Hz, 2H),
2.82 (dd, J = 14.7, 6.5 Hz, 1H), 2.74 (dd, J = 13.6, 5.7 Hz, 1H), 2.61
(dd, J = 13.8, 7.8 Hz, 1H), 2.41 (d, J = 15.8 Hz, 1H), 2.22–2.17 (m,
1
4
28.2, 127.4, 126.4, 124.5, 122.4, 108.5, 74.3, 68.5, 67.0, 65.4, 48.7,
3.2, 41.8, 41.2, 27.9, 21.8, 20.4, 19.1, 15.7, 15.6; MS m/z: [M+Na]
+
6
calc’d for C32H45NO Na 562.3; found 562.4.
(E)-3-(3-((S)-2-(diethoxymethyl)-2-methyl-4-oxo-4-(((S)-1-
phenylethyl)amino)butyl)phenyl)allyl isobutyl carbonate (S9)
Synthesized according to General Procedure A using (S)-(-)-
phenylethylamine and (+)-S5 [47 µmol]. Obtained 22 mg [crude,
8
7%] of (E)-3-(3-((S)-2-(diethoxymethyl)-2-methyl-4-oxo-4-(((S)-1-
phenylethyl)amino)butyl)phenyl)allyl isobutyl carbonate. The crude
2H), 2.13–2.00 (m, 3H),1.90 (quint., J = 7.0 Hz, 1H), 1.86–1.80 (m,
1H), 1.75–1.65 (m, 1H), 1.42 (s, 3H), 0.89 (d, J = 6.8 Hz, 6H);
1
13
was then dissolved in 500 µL of CDCl
3
for H-NMR analysis of dia-
, 500 MHz): δ 7.34–7.32 (m,
H), 7.28–7.23 (m, 3H), 7.19 (dd, J = 7.6, 7.6 Hz, 1H), 7.12 (ddd, J =
C
1
stereomeric mixture. H NMR (CDCl
3
NMR (DMSO-d , 126 MHz): δ174.1, 172.6, 172.5, 170.6, 170.2,
6
4
7
6
155.8, 154.5, 137.6, 136.8, 135.4, 133.6, 130.4, 130.1, 129.1, 128.8,
128.2, 127.5, 126.4, 124.5, 123.2, 121.2, 118.7, 117.9, 114.9, 111.4,
107.1, 73.4, 67.7, 62.7, 53.9, 52.2, 49.3, 42.6, 42.1, 36.6, 30.2, 27.3,
.5, 1.2, 1.2 Hz, 1H), 6.64 (d, J = 15.9 Hz, 1H), 6.27 (ddd, J = 15.8,
.5, 6.5 Hz, 1H), 6.01 (d, J = 7.8 Hz, 1H), 5.12 (quint., J = 7.1 Hz,
+
1H), 4.77 (dd, J = 6.5, 1.0 Hz, 2H), 4.25 (s, 1H), 3.94 (d, J = 6.7 Hz,
H), 3.82–3.69 (m, 2H), 3.51–3.44 (m, 1H), 3.42–3.36 (m, 1H), 2.87
2.76 (AB quartet, J = 13.0 Hz, 2H), 2.23 & 2.09 (AB quartet, J =
26.8, 23.5, 23.2, 18.7; HRMS (QE Orbitrap) m/z: [M+H] calc’d for
2
&
C
45
H
52
N
5
O
10 822.3709; found 822.3676.
3-((8S,12S,15S,18S,E)-12-((1H-indol-3-yl)methyl)-18-
1
7
3.9 Hz, 2H), 2.03–1.94 (m, 1H), 1.48 (d, J = 7.0 Hz, 3H), 1.24 (t, J =
.0 Hz, 3H), 1.15 (t, J = 7.0 Hz, 3H), 0.96 (d, J = 6.7 Hz, 6H), 0.95 (s,
carbamoyl-8-(diethoxymethyl)-8-methyl-10,13,16-trioxo-2-oxa-
11,14,17-triaza-1(1,4),6(1,3)-dibenzenacyclononadecaphan-4-en-
15-yl)propanoic acid (S16)
1
3
3H); C NMR (CDCl
3
, 126 MHz): δ 171.1, 155.4, 143.6, 138.7,
35.7, 135.1, 131.3, 129.7, 128.8, 128.2, 127.5, 126.4, 124.5, 122.4,
08.6, 74.3, 68.5, 66.7, 65.8, 48.7, 43.1, 41.8, 41.0, 27.9, 21.7, 20.7,
1
1
1
Synthesized according to Procedure E beginning with 53 mg of
crude 28. Carried forward without purification.
+
1
5
11b
1
3
9.0, 15.7, 15.6; MS m/z: [M+Na] calc’d for C32
H
45NO
6
Na 562.3;
3-((1 S,1 S,1 R,10S,13S,E)-10-carbamoyl-1 -methyl-1 ,12,15-
2 3 5 6 11 11b 1
trioxo-1 ,1 ,1 ,1 ,1 ,1 -hexahydro-1 H-7-oxa-11,14-diaza-1(1,5)-
found 562.4.
ACS Paragon Plus Environment

The Journal of Organic Chemistry
Page 12 of 20
indolizino[8,7-b]indola-3(1,3),8(1,4)-
Synthesized according to Procedure A beginning with 0.20 mmol
1
2
3
4
5
6
7
8
9
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
2
2
2
3
3
3
3
3
3
3
3
3
3
4
4
4
4
4
4
4
4
4
4
5
5
5
5
5
5
5
5
5
5
6
dibenzenacyclopentadecaphan-4-en-13-yl)propanoic acid (30)
Synthesized according to Procedure B using crude material from
the previous reaction. Purified by preparative HPLC – see SI for con-
of (+)-S5. Carried forward without purification.
(S)-5-(((S)-1-amino-3-(4-hydroxyphenyl)-1-oxopropan-2-
yl)amino)-4-((1S,5R,11bS)-1-(3-((E)-3-
((isobutoxycarbonyl)oxy)prop-1-en-1-yl)benzyl)-1-methyl-3-oxo-
2,3,5,6,11,11b-hexahydro-1H-indolizino[8,7-b]indole-5-
carboxamido)-5-oxopentanoic acid (S18)
ditions. White Solid. 11.4 mg [16 µmol, 28% yield over three steps].
1
6
H NMR (DMSO-d , 500 MHz): δ 12.04 (br s, 1H), 10.87 (s, 1H),
8.32 (d, J = 9.2 Hz, 1H), 7.41 (d, J = 8.2 Hz, 1H), 7.38 (br s, 1H),
7
6
6
.25 (d, J = 8.7 Hz, 2H), 7.11–7.01 (m, 5H), 7.04 (d, J = 8.9 Hz, 2H),
.98–6.95 (m, 1H), 6.92 (d, J = 6.8 Hz, 1H), 6.75 (d, J = 8.7 Hz, 1H),
.54 (d, J = 16.3 Hz, 1H), 6.25 (s, 1H), 5.98 (ddd, J = 16.3, 6.8, 4.0
Synthesized according to Procedure B using the crude from the
previous reaction. Carried forward without purification.
1
5
11b
6
1
3-((1 S,1 R,1 S,9S,12S,E)-9-carbamoyl-7 -hydroxy-1 -methyl-
3
2
3
5
6
11 11b
1
Hz, 1H), 4.93 (s, 1H), 4.87 (ddd, J = 15.9, 4.0, 1.7 Hz, 1H), 4.80 (dd,
J = 15.9, 6.8 Hz, 1H), 4.48–4.44 (m, 1H), 4.32 (d, J = 9.2 Hz, 1H),
1 ,11,14-trioxo-1 ,1 ,1 ,1 ,1 ,1 -hexahydro-1 H-10,13-diaza-
1(1,5)-indolizino[8,7-b]indola-3,7(1,3)-
4
2
.13 (ddd, J = 9.1, 9.1, 4.8 Hz, 1H), 3.11 (dd, J = 14.5, 5.3 Hz, 1H),
.76 (dd, J = 14.5, 4.3 Hz, 1H), 2.18 (d, J = 16.8 Hz, 1H), 2.10–2.07
dibenzenacyclotetradecaphan-4-en-12-yl)propanoic acid (34)
Synthesized according to Procedure C using 6 mg of crude S18.
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
(
m, 2H), 2.01–1.94 (m, 1H), 1.82–1.76 (m, 1H), 1.64 (s, 3H), 1.62–
Purified by preparative HPLC – see SI for conditions. White Solid. 3
13
1
1
.56 (m, 1H), 1.54–1.48 (m, 1H), 1.41–1.35 (m, 1H); C NMR
(DMSO-d , 126 MHz): δ 174.4, 174.0, 172.2, 171.9, 171.6, 170.1,
157.6, 137.5, 137.0, 134.5, 131.3, 130.7, 130.0, 129.3, 128.1, 127.7,
mg [4.3 µmol, 60% yield over three steps]. H NMR (DMSO-d
6
, 500
6
MHz): δ 12.06 (br s, 1H), 10.64 (br s, 1H), 9.18 (s, 1H), 8.08 (d, J =
4.8 Hz, 1H), 7.42 (s, 1H), 7.41 (d, J = 8.1 Hz, 1H), 7.37 (br s, 1H),
7.36 (d, J = 7.4 Hz, 2H), 7.35 (dd, J = 7.3, 7.3 Hz, 1H), 7.35 (d, J =
7.6 Hz, 1H), 7.29 (d, J = 7.7 Hz, 1H), 7.10 (d, J = 7.2 Hz, 1H), 7.08
(dd, J = 7.9, 7.9 Hz, 2H), 6.98 (dd, J = 7.7, 7.7 Hz, 1H), 6.95 (br s,
1H), 6.86 (d, J = 1.7 Hz, 1H), 6.84 (dd, J = 8.2, 1.8 Hz, 1H), 6.66 (d,
J = 8.1 Hz, 1H), 6.52 (d, J = 15.7 Hz, 1H), 6.32 (ddd, J = 15.6, 7.2,
7.2 Hz, 1H), 5.51 (s, 1H), 5.03 (d, J = 6.6 Hz, 1H), 4.19 (ddd, J = 8.1,
8.1, 3.8 Hz, 1H), 3.81–3.77 (m, 1H), 3.40 (dd, J = 15.6, 7.8 Hz, 1H),
3.39 (d, J = 13.8 Hz, 1H), 3.24 (dd, J = 15.5, 6.6 Hz, 1H), 3.15 (d, J
= 15.2 Hz, 1H), 2.88 (dd, J = 15.2, 6.5 Hz, 1H), 2.78 (d, J = 13.8 Hz,
1H), 2.66 (dd, J = 13.9, 3.5 Hz, 1H), 2.58 (dd, J = 13.9, 8.4 Hz, 1H),
2.29 & 1.90 (AB quartet, J = 16.1 Hz, 2H), 2.15 (ddd, J = 16.3, 11.1,
5.3 Hz, 1H), 2.06 (ddd, J = 16.4, 11.1, 5.2 Hz, 1H), 1.83–1.77 (m,
1
6
27.2, 126.5, 126.0, 125.0, 121.2, 118.7, 118.4, 115.0, 111.1, 106.0,
8.4, 52.0, 50.8, 48.6, 47.7, 43.2, 42.4, 36.3, 32.0, 27.1, 26.2, 23.7;
+
HRMS (QE Orbitrap) m/z: [M+Na] calc’d for C40
found 704.3071. *Contaminated with an unknown impurity. C-NMR
peaks chosen by analogy to Macrocycle 35*
H
41
N
5
O
7
704.3079;
1
3
(
5S,9S,19S)-9-((1H-indol-3-yl)methyl)-19-(((S)-1-amino-3-(4-
hydroxyphenyl)-1-oxopropan-2-yl)carbamoyl)-4-ethoxy-5-(3-((E)-
3
7
-((isobutoxycarbonyl)oxy)prop-1-en-1-yl)benzyl)-5-methyl-
,10,17-trioxo-3-oxa-8,11,18-triazadocosan-22-oic acid (31)
Synthesized according to Procedure A beginning with 0.22 mmol
of (+)-S5. Carried forward without purification.
S)-5-(((S)-1-amino-3-(4-hydroxyphenyl)-1-oxopropan-2-
yl)amino)-4-(6-((1S,5S,11bR)-1-(3-((E)-3-
(isobutoxycarbonyl)oxy)prop-1-en-1-yl)benzyl)-1-methyl-3-oxo-
,3,5,6,11,11b-hexahydro-1H-indolizino[8,7-b]indole-5-
(
1
3
6
1H), 1.68–1.62 (m, 1H), 0.81 (s, 3H); C NMR (DMSO-d , 151
(
2
MHz): δ 173.9, 173.0, 171.1, 170.3, 153.3, 137.7, 137.7, 136.3, 130.8,
130.3, 130.0, 129.9, 129.3, 128.3, 128.1, 127.8, 127.7, 126.1, 125.4,
124.2, 120.8, 118.4, 117.1, 114.3, 111.1, 105.8, 57.0, 53.9, 52.1, 48.8,
carboxamido)hexanamido)-5-oxopentanoic acid (S17)
Synthesized according to Procedure B using crude material from
the previous reaction. Carried forward without purification.
41.9, 41.8, 41.2, 36.8, 32.4, 30.2, 26.7, 25.3, 23.7; HRMS (QE Or-
+
bitrap) m/z: [M+H] calc’d for C40
H
42
N
5
O
7
704.3079; found 704.3072.
1
5
11b
1
1
5
11b
1
3
3
-((1 S,1 S,1 R,10S,13S,E)-10-carbamoyl-1 -methyl-
3-((1 S,1 R,1 S,10S,13S,E)-10-carbamoyl-1 -methyl-1 ,12,15-
3
2
3
5
6
11 11b
1
2 3 5 6 11 11b 1
1
,12,15,22-tetraoxo-1 ,1 ,1 ,1 ,1 ,1 -hexahydro-1 H-7-oxa-
trioxo-1 ,1 ,1 ,1 ,1 ,1 -hexahydro-1 H-7-oxa-11,14-diaza-1(1,5)-
indolizino[8,7-b]indola-3(1,3),8(1,4)-
11,14,21-triaza-1(1,5)-indolizino[8,7-b]indola-3(1,3),8(1,4)-
dibenzenacyclodocosaphan-4-en-13-yl)propanoic acid (32)
Synthesized according to Procedure D using 32 mg of crude S17.
dibenzenacyclopentadecaphan-4-en-13-yl)propanoic acid (35)
Synthesized according to Procedure D using 50 mg of crude S18.
Purified by preparative HPLC using HCOOH as modifier – see SI for
Purified by preparative HPLC – see SI for conditions. White Solid. 9
1
mg [11 µmol, 19% yield over three steps]. H NMR (DMSO-d
6
, 600
conditions. White Solid. 13 mg [18 µmol, 29% yield over three steps].
1
MHz): δ 10.87 (s, 1H), 7.95 (d, J = 8.2 Hz, 1H), 7.74 (dd, J = 5.6, 5.6
Hz, 1H), 7.63 (d, J = 8.1 Hz, 1H), 7.50 (br s, 1H), 7.40 (d, J = 7.9 Hz,
6
H NMR (DMSO-d , 600 MHz): δ 12.11 (br s, 1H), 11.05 (s, 1 H),
8.12 (d, J = 4.2 Hz, 1 H), 7.52 (d, J = 5.6 Hz, 1H), 7.52 (d, J = 5.6
Hz, 1 H), 7.40 (br s, 1H), 7.37 (d, J = 6.7 Hz, 1H), 7.36-7.34 (m, 1H),
7.34 (s, 1 H), 7.16 (d, J = 6.2 Hz, 2H), 7.13 (d, J = 6.7 Hz, 1 H), 7.11
(dd, J = 6.9, 6.9 Hz, 1H), 7.01 (d, J = 5.7 Hz, 1 H), 7.00 (dd, J = 6.7,
6.7 Hz, 1H), 6.90 (br s, 1H), 6.83 (d, J = 6.1 Hz, 2H), 6.71 (d, J =
15.7 Hz, 1 H), 6.38-6.36 (m, 1 H), 5.19 (s, 1 H), 5.08 (d, J = 4.1 Hz, 1
H), 4.92 (dd, J = 14.6, 5.0 Hz, 1H), 4.81 (d, J = 14.5 Hz, 1H), 4.09-
4.07 (m, 1H), 3.70-3.67 (m, 1 H), 3.39 & 2.73 (AB quartet, J = 13.8
Hz, 2 H), 3.07 (d, J = 15.3 Hz, 1H), 2.90 (dd, J = 15.3, 4.2 Hz, 1H),
2.83 (d, J = 13.8 Hz, 1H), 2.60 (dd, J = 13.9, 9.2 Hz, 1H), 2.28-2.24
(m, 1H), 2.25 & 1.88 (AB quartet, J = 15.3 Hz, 2 H), 2.17-2.13 (m,
1
=
H), 7.32 (d, J = 7.9 Hz, 1H), 7.25 (d, J = 8.6, Hz, 1H), 7.17 (ddd, J
7.7, 1.3, 1.3 Hz, 1H), 7.12 (dd, J = 7.5, 7.5 Hz, 1H), 7.08 (br s, 1H),
6
8
.96 (ddd, J = 7.9, 7.0, 0.8 Hz, 1H), 6.95–6.94 (m, 1H), 6.93 (d, J =
.7 Hz, 1H), 6.88 (ddd, J = 7.9, 7.0, 0.8 Hz, 1H), 6.31 (dd, J = 1.4,
1.4 Hz, 1H), 6.23 (d, J = 16.1 Hz, 1H), 6.10 (ddd, J = 16.0, 5.8, 5.8
Hz, 1H), 5.01 (dd, J = 1.4, 1.4 Hz, 1H), 4.74 (d, J = 7.0 Hz, 1H), 4.65
(
m, 2H), 4.40 (ddd, J = 9.9, 8.2, 3.4 Hz, 1H), 4.31 (ddd, J = 7.9, 7.9,
5
1
.7 Hz, 1H), 3.21 (d, J = 15.1 Hz, 1H), 2.98 (dd, J = 14.0, 3.6 Hz,
H), 2.96–2.93 (m, 2H), 2.78 (dd, J = 14.0, 10.0 Hz, 1H), 2.58 (ddd, J
= 15.3, 6.3, 1.6 Hz, 2H), 2.51 & 2.37 (AB quartet, J = 15.8Hz, 2H),
2.39 (s, 2H), 2.24–2.15 (m, 2H), 1.98 (ddd, J = 14.9, 9.5, 5.7 Hz, 1H),
1
3
1H), 1.90-1.85 (m, 1H), 1.70-1.65 (m, 1 H), 0.74 (s, 3 H); C NMR
(DMSO-d , 151 MHz): δ 173.8, 173.1, 171.3, 170.9, 170.6, 155.7,
1
.92–1.86 (m, 1H), 1.76 (ddd, J = 15.0, 9.5, 5.5 Hz, 1H), 1.74–1.69
6
(
m, 1H), 1.45 (s, 3H), 1.37–1.31 (m, 1H), 1.29–1.24 (m, 1H), 1.22–
137.5, 136.3, 135.7, 132.2, 131.2, 130.2, 130.1, 130.0, 129.6, 128.4,
126.1, 125.9, 123.0, 120.6, 118.5, 117.0, 114.8, 111.1, 105.3, 66.9,
13
1
.17 (m, 1H), 1.17–1.13 (m, 1H), 0.93–0.87 (m, 2H); C NMR
DMSO-d , 151 MHz): δ 173.9, 173.0, 172.0, 171.9, 171.0, 169.7,
156.8, 137.3, 136.6, 135.0, 132.1, 130.2, 129.9, 129.4, 129.3, 128.1,
(
6
56.8, 54.3, 48.3, 48.2, 42.0, 41.2, 40.9, 35.8, 30.3, 26.4, 26.0, 23.5;
+
HRMS (QE Orbitrap) m/z: [M+H] calc’d for C40
H
42
N
5
O
7
704.3079;
1
6
2
27.7, 126.2, 124.4, 124.0, 121.0, 118.4, 117.5, 114.1, 111.1, 106.7,
7.6, 62.1, 54.1, 51.2, 49.1, 43.7, 42.3, 40.9, 37.7, 36.4, 35.0, 29.8,
8.1, 27.6, 25.6, 25.3, 24.6, 23.4; HRMS (QE Orbitrap) m/z: [M+H]
found 704.3057.
(S)-5-(((S)-1-amino-3-(4-hydroxyphenyl)-1-oxopropan-2-
yl)amino)-4-((R)-2-((S)-3-(diethoxymethyl)-3-(3-((E)-3-
((isobutoxycarbonyl)oxy)prop-1-en-1-yl)benzyl)oct-7-ynamido)-3-
(1H-indol-3-yl)propanamido)-5-oxopentanoic acid (S19)
Synthesized according to Procedure A beginning with 0.57 mmol
of (+)-5. Carried forward without purification.
(S)-5-(((S)-1-amino-3-(4-hydroxyphenyl)-1-oxopropan-2-
yl)amino)-4-((1S,5R,11bS)-1-(3-((E)-3-
((isobutoxycarbonyl)oxy)prop-1-en-1-yl)benzyl)-3-oxo-1-(pent-4-
+
53 6 8
calc’d for C46H N O 817.3919; found 817.3927.
(S)-5-(((S)-1-amino-3-(4-hydroxyphenyl)-1-oxopropan-2-
yl)amino)-4-((R)-2-((S)-4,4-diethoxy-3-(3-((E)-3-
(
(isobutoxycarbonyl)oxy)prop-1-en-1-yl)benzyl)-3-
methylbutanamido)-3-(1H-indol-3-yl)propanamido)-5-
oxopentanoic acid (33)
ACS Paragon Plus Environment

Page 13 of 20
The Journal of Organic Chemistry
1
3
yn-1-yl)-2,3,5,6,11,11b-hexahydro-1H-indolizino[8,7-b]indole-5-
carboxamido)-5-oxopentanoic acid (S20)
(m, 1H), 1.37–1.32 (m, 1H), 1.25–1.21 (m, 1H); C NMR (DMSO-
6
d , 151 MHz): δ 173.9, 173.0, 171.4, 171.1, 170.2, 153.2, 137.7,
1
2
3
4
5
6
7
8
9
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
2
2
2
3
3
3
3
3
3
3
3
3
3
4
4
4
4
4
4
4
4
4
4
5
5
5
5
5
5
5
5
5
5
6
Synthesized according to Procedure B using the crude material
from the previous reaction. Carried forward without purification.
137.4, 136.4, 130.7, 130.1, 129.9, 129.9, 129.4, 128.6, 128.4, 127.8,
127.6, 126.0, 125.4, 124.0, 120.8, 118.5, 117.3, 114.4, 111.1, 106.1,
83.9, 71.0, 57.4, 54.1, 52.6, 52.6, 48.8, 44.3, 39.9, 38.4, 36.8, 34.6,
32.2, 31.4, 25.0, 22.5, 18.1; HRMS (DART-Orbitrap) m/z: [M+H]
47 6 6
calc’d for C44H N O 755.3552; found 755.3576.
(E)-3-(3-((3S,6S,13R,17S)-13-((1H-indol-3-yl)methyl)-6-(((S)-1-
amino-3-(4-hydroxyphenyl)-1-oxopropan-2-yl)carbamoyl)-17-
(diethoxymethyl)-3-methyl-1,4,12,15-tetraoxo-17-(pent-4-yn-1-yl)-
1-((S)-pyrrolidin-2-yl)-2,5,11,14-tetraazaoctadecan-18-
yl)phenyl)allyl isobutyl carbonate (S23)
1
5
11b
6
3
3
-((1 S,1 R,1 S,9S,12S,E)-9-carbamoyl-7 -hydroxy-1 ,11,14-
1 2 3 5 6 11 11b 1
+
trioxo-1 -(pent-4-yn-1-yl)-1 ,1 ,1 ,1 ,1 ,1 -hexahydro-1 H-10,13-
diaza-1(1,5)-indolizino[8,7-b]indola-3,7(1,3)-
dibenzenacyclotetradecaphan-4-en-12-yl)propanoic acid (36)
Synthesized according to Procedure C using 280 mg of crude mate-
rial from the previous reaction. Purified by SiO
à10% MeOH/CHCl (0.1% AcOH). White Solid. 133 mg [176
µmol, 31% yield over three steps]. H NMR (DMSO-d
2
chromatography
1
3
1
6
, 600 MHz): δ
2.06 (br s, 1H), 10.59 (s, 1H), 9.17 (s, 1H), 8.05 (d, J = 6.6 Hz, 1H),
.42 (d, J = 7.9 Hz, 1H), 7.37 (s, 1H), 7.37 (d, J = 7.8 Hz, 1H), 7.37
1
7
Synthesized according to Procedure A beginning with 0.43 mmol
of (+)-5. Carried forward without purification.
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
(
7
br s, 1H), 7.35 (dd, J = 7.4, 7.4 Hz, 1H), 7.35 (d, J = 8.1 Hz, 1H),
.31 (d, J = 7.7 Hz, 1H), 7.10 (d, J = 7.7 Hz, 1H), 7.08 (ddd, J = 7.6,
(E)-3-(3-(((1S,5R,11bS)-5-(((S)-6-(((S)-1-amino-3-(4-
hydroxyphenyl)-1-oxopropan-2-yl)amino)-6-oxo-5-((S)-2-((S)-
pyrrolidine-2-carboxamido)propanamido)hexyl)carbamoyl)-3-
oxo-1-(pent-4-yn-1-yl)-2,3,5,6,11,11b-hexahydro-1H-
indolizino[8,7-b]indol-1-yl)methyl)phenyl)allyl isobutyl carbonate
(S24)
7.6, 0.8 Hz, 1H), 6.98 (dd, J = 7.6, 7.6 Hz, 1H), 6.95 (br s, 1H), 6.86
(d, J = 1.6 Hz, 1H), 6.84 (dd, J = 8.3, 1.9 Hz, 1H), 6.66 (d, J = 8.1 Hz,
1
5
1
1
H), 6.49 (d, J = 15.7 Hz, 1H), 6.33 (ddd, J = 15.6, 7.2, 7.2 Hz, 1H),
.60 (s, 1H), 5.02 (d, J = 6.6 Hz, 1H), 4.20 (ddd, J = 8.2, 8.2, 3.8 Hz,
H), 3.80 (ddd, J = 9.3, 6.7, 4.4 Hz, 1H), 3.50 (dd, J = 15.6, 7.3 Hz,
H), 3.34–3.32 (m (under water), 1H), 3.21 (dd, J = 15.6, 6.9Hz, 1H),
Synthesized according to Procedure B using 368 mg of crude S23.
Carried forward without purification.
1
5
11b
6
3
1
3.17 (d, J = 15.5 Hz, 1H), 3.17 (d, J = 15.4 Hz, 1H), 2.87–2.83 (m,
(1 S,1 R,1 S,9S,12S,E)-7 -hydroxy-1 ,11,18-trioxo-1 -(pent-4-
yn-1-yl)-12-((S)-2-((S)-pyrrolidine-2-carboxamido)propanamido)-
2
2
H), 2.66 (dd, J = 13.9, 3.4 Hz, 1H), 2.59 (dd, J = 14.0, 8.3 Hz, 1H),
.56 (dd, J = 2.6, 2.6 Hz, 1H), 2.19 (d, J = 16.4 Hz, 1H), 2.15 (ddd, J
2
3
5
6
11 11b
1
1 ,1 ,1 ,1 ,1 ,1 -hexahydro-1 H-10,17-diaza-1(1,5)-
indolizino[8,7-b]indola-3,7(1,3)-dibenzenacyclooctadecaphan-4-
ene-9-carboxamide (38)
=
1
16.3, 11.3, 5.1 Hz, 1H), 2.09 (d, J = 16.4 Hz, 1H), 2.05 (ddd, J =
6.4, 11.3, 5.1 Hz, 1H), 1.91 (ddd, J = 6.6, 6.6, 2.5 Hz, 2H), 1.83–
1.78 (m, 1H), 1.68–1.62 (m, 1H), 1.47–1.40 (m, 1H), 1.38–1.33 (m,
Synthesized according to Procedure C using crude material from
the previous reaction. Purified by preparative HPLC – see SI for
1
3
6
1H), 1.36–1.33 (m, 1H), 1.26–1.23 (m, 1H); C NMR (DMSO-d ,
1
1
1
7
51 MHz): δ 173.7, 172.7, 171.1, 171.0, 170.0, 153.2, 137.6, 137.5,
36.4, 130.5, 130.0, 130.0, 129.8, 129.5, 128.6, 128.4, 127.7, 127.5,
26.1, 125.4, 123.7, 120.5, 118.3, 116.9, 114.3, 110.9, 106.0, 83.8,
conditions. Yellow solid. HCl-salt: 47 mg [49 µmol, 13% yield over
1
three steps]. H NMR (DMSO-d
6
, 600 MHz): δ 10.50 (s, 1H), 9.82 (br
s, 1H), 8.73 (d, J = 7.1 Hz, 1H), 8.46–8.43 (m, 1H), 8.18 (d, J = 7.3
Hz, 1H), 7.60 (d, J = 7.2 Hz, 1H), 7.56 (dd, J = 5.9, 5.9 Hz, 1H), 7.54
(s, 1H), 7.42 (d, J = 8.1 Hz, 1H), 7.40 (d, J = 8.2 Hz, 1H), 7.37 (br s,
1H), 7.33–7.31 (m, 1H), 7.31 (dd, J = 7.7, 7.7 Hz, 1H), 7.20 (br s,
1H), 7.15–7.13 (m, 1H), 7.10 (dd, J = 8.1, 8.1 Hz, 1H), 7.00 (dd, J =
7.9, 7.9 Hz, 1H), 6.93 (d, J = 1.6 Hz, 1H), 6.84 (dd, J = 8.3, 1.6 Hz,
1H), 6.68 (d, J = 8.2 Hz, 1H), 6.46 (ddd, J = 15.7, 6.6, 6.6 Hz, 1H),
6.35 (d, J = 15.9 Hz, 1H), 4.96 (s, 1H), 4.88 (d, J = 6.6 Hz, 1H), 4.39–
4.34 (m, 1H), 4.33 (ddd, J = 7.0, 7.0, 7.0 Hz, 1H), 4.14 (dddd, J = 6.0,
6.0, 6.0, 6.0 Hz, 1H), 3.97 (ddd, J = 8.1, 8.1, 5.3 Hz, 1H), 3.47–3.44
(under water) (m, 1H), 3.46–3.43 (under water) (m, 1H), 3.35 (dd, J =
16.1, 6.9 Hz, 1H), 3.29 & 2.99 (AB quartet, J = 13.7 Hz, 2H), 3.18–
3.12 (m, 2H), 2.94 (dd, J = 13.9, 6.1 Hz, 1H), 2.91 (dd, J = 14.0, 4.0
Hz, 1H), 2.91 & 2.06 (AB quartet, J = 16.1 Hz, 2H), 2.78 (dd, J =
14.0, 8.9 Hz, 1H), 2.66 (dd, J = 15.7, 6.5 Hz, 1H), 2.66 (dd, J = 14.1,
5.8 Hz, 1H), 2.52 (dd, J = 2.4, 2.4 Hz, 1H), 2.28–2.22 (m, 1H), 1.88–
1.86 (m, 2H), 1.84–1.78 (m, 2H), 1.82–1.78 (m, 1H), 1.62–1.58 (m,
1H), 1.42–1.35 (m, 1H), 1.32–1.28 (m, 2H), 1.32–1.28 (m, 1H), 1.25–
0.8, 57.3, 53.9, 52.0, 48.7, 44.1, 39.8, 38.1, 36.7, 34.7, 32.4, 30.0,
+
26.8, 25.1, 22.6, 18.1; HRMS (QE Orbitrap) m/z: [M+H] calc’d for
C
44
H
46
N
5
O
7
756.3392; found 756.3391.
(
E)-3-(3-((S)-2-(2-(((R)-1-(((S)-5-amino-1-(((S)-1-amino-3-(4-
hydroxyphenyl)-1-oxopropan-2-yl)amino)-1,5-dioxopentan-2-
yl)amino)-3-(1H-indol-3-yl)-1-oxopropan-2-yl)amino)-2-oxoethyl)-
2-(diethoxymethyl)hept-6-yn-1-yl)phenyl)allyl isobutyl carbonate
(
S21)
Synthesized according to Procedure A beginning with 0.22 mmol
of (+)-5. Carried forward without purification.
E)-3-(3-(((1S,5R,11bS)-5-(((S)-5-amino-1-(((S)-1-amino-3-(4-
(
hydroxyphenyl)-1-oxopropan-2-yl)amino)-1,5-dioxopentan-2-
yl)carbamoyl)-3-oxo-1-(pent-4-yn-1-yl)-2,3,5,6,11,11b-hexahydro-
1
H-indolizino[8,7-b]indol-1-yl)methyl)phenyl)allyl isobutyl car-
bonate (S22)
Synthesized according to Procedure B using 162 mg of crude S21.
Carried forward without purification.
1
5
11b
6
(1 S,1 R,1 S,9S,12S,E)-12-(3-amino-3-oxopropyl)-7 -hydroxy-
1.21 (m, 1H), 1.24 (d, J = 7.0 Hz, 3H), 1.06–1.01 (m, 2H), 1.06–0.96
3
1
1
2
3
5
6
11 11b
13
1
1
,11,14-trioxo-1 -(pent-4-yn-1-yl)-1 ,1 ,1 ,1 ,1 ,1 -hexahydro-
6
(m, 2H); C NMR (DMSO-d , 151 MHz): δ 172.6, 172.2, 171.7,
H-10,13-diaza-1(1,5)-indolizino[8,7-b]indola-3,7(1,3)-
170.7, 168.1, 167.5, 153.1, 137.5, 137.4, 136.5, 130.4, 129.5, 129.5,
129.3, 128.9, 128.5, 128.0, 127.4, 127.3, 126.0, 125.0, 124.0, 120.9,
118.4, 117.6, 114.3, 111.1, 106.9, 83.6, 70.9, 58.3, 57.4, 53.4, 53.2,
48.9, 48.3, 45.3, 44.6, 40.5, 39.1, 38.0, 36.3, 34.3, 32.2, 30.7, 29.3,
dibenzenacyclotetradecaphan-4-ene-9-carboxamide (37)
Synthesized according to Procedure C using the crude material
from the previous reaction. Purified by preparative HPLC – see SI for
conditions. White Solid. 27 mg [36 µmol, 22% yield over three steps].
27.8, 23.3, 22.3, 22.2, 21.5, 18.0, 17.5; HRMS (QE Orbitrap) m/z:
1
+
H NMR (DMSO-d
6
, 600 MHz): δ 10.51 (s, 1H), 9.20 (br s, 1H), 8.18
[M+H] calc’d for C53
H
63
N
8
O
7
923.4814; found 923.4769.
(
d, J = 6.3 Hz, 1H), 7.41 (d, J = 7.8 Hz, 1H), 7.40 (s, 1H), 7.38 (br s,
(S)-4-((R)-2-((S)-3-(diethoxymethyl)-3-(3-((E)-3-
((isobutoxycarbonyl)oxy)prop-1-en-1-yl)benzyl)oct-7-ynamido)-3-
(1H-indol-3-yl)propanamido)-5-((4-hydroxyphenethyl)amino)-5-
oxopentanoic acid (S25)
Synthesized according to Procedure A beginning with 0.26 mmol
of (+)-5. Carried forward without purification.
(S)-5-((4-hydroxyphenethyl)amino)-4-((1S,5R,11bS)-1-(3-((E)-
3-((isobutoxycarbonyl)oxy)prop-1-en-1-yl)benzyl)-3-oxo-1-(pent-
4-yn-1-yl)-2,3,5,6,11,11b-hexahydro-1H-indolizino[8,7-b]indole-5-
carboxamido)-5-oxopentanoic acid (S26)
Synthesized according to Procedure B using crude material from
the previous reaction. Carried forward without purification.
1
1
H), 7.37–7.34 (m, 1H), 7.34 (d, J = 7.9 Hz, 1H), 7.33 (d, J = 6.4 Hz,
H), 7.31 (d, J = 7.5 Hz, 1H), 7.22 (d, J = 6.8 Hz, 1H), 7.11 (d, J =
8.1 Hz, 1H), 7.08 (dd, J = 8.1, 8.1 Hz, 1H), 7.00 (br s, 2H), 6.98 (dd, J
7.6, 7.6 Hz, 1H), 6.85 (br s, 1H) , 6.84 (d, J = 8.5 Hz, 1H), 6.67 (d,
J = 8.7 Hz, 1H), 6.49 (d, J = 15.6 Hz, 1H), 6.35 (ddd, J = 15.6, 7.1,
=
7
1
.1 Hz, 1H), 5.59 (s, 1H), 5.03 (d, J = 6.4 Hz, 1H), 4.20–4.16 (m,
H), 3.79–3.75 (m, 1H), 3.48 (dd, J = 15.8, 7.3 Hz, 1H), 3.33 & 2.83
(AB quartet, J = 13.9 Hz, 2H), 3.24 (dd, J = 15.8, 6.8 Hz, 1H), 3.20
d, J = 15.9 Hz, 1H), 2.85 (dd, J = 15.9, 6.4 Hz, 1H), 2.66 (dd, J =
3.8, 2.5 Hz, 1H), 2.58 (dd, J = 2.5, 2.5 Hz, 1H), 2.54–2.50 (under
DMSO) (m, 2H), 2.22 & 2.11 (AB quartet, J = 16.5 Hz, 2H), 2.08–
.01 (m, 1H), 1.98–1.93 (m, 1H), 1.92 (ddd, J = 6.1, 6.1, 2.3 Hz, 2H),
1.80–1.74 (m, 1H), 1.66–1.59 (m, 1H), 1.48–1.41 (m, 1H), 1.37–1.32
(
1
2
ACS Paragon Plus Environment

The Journal of Organic Chemistry
Page 14 of 20
3
1
5
11b
6
1
Synthesized according to Procedure C using 55mg of crude S28.
Purified by preparative HPLC – see SI for conditions. Pale yellow
3
-((1 S,1 R,1 S,12S,E)-7 -hydroxy-1 ,11,14-trioxo-1 -(pent-4-
2 3 5 6 11 11b 1
1
2
3
4
5
6
7
8
9
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
2
2
2
3
3
3
3
3
3
3
3
3
3
4
4
4
4
4
4
4
4
4
4
5
5
5
5
5
5
5
5
5
5
6
yn-1-yl)-1 ,1 ,1 ,1 ,1 ,1 -hexahydro-1 H-10,13-diaza-1(1,5)-
indolizino[8,7-b]indola-3,7(1,3)-dibenzenacyclotetradecaphan-4-
en-12-yl)propanoic acid (39)
1
solid. 27 mg [37 µmol, 57% yield over three steps]. H NMR
DMSO-d , 600 MHz): δ 11.01 (s, 1H), 9.28 (br s, 1H), 7.81 (d, J =
.0 Hz, 1H), 7.45 (dd, J = 5.3, 5.3 Hz, 1H), 7.44 (s, 1H), 7.39 (d, J =
.9 Hz, 1H), 7.36 (d, J = 7.9 Hz, 1H), 7.34 (dd, J = 7.2, 7.2 Hz, 1H),
(
6
7
7
Synthesized according to Procedure C using 50 mg of crude S26.
Purified by preparative TLC: 3% MeOH / 96.5%CHCl
White solid. 20 mg [28 µmol, 57% yield over three steps]. H NMR
(DMSO-d , 600 MHz): δ 10.66 (s, 1H), 9.21 (br s, 1H), 7.45 (s, 1H),
.37 (dd, J = 5.7, 5.7 Hz, 1H), 7.37 (d, J = 7.3 Hz, 1H), 7.35 (dd, J =
.8, 5.8 Hz, 1H), 7.35 (d, J = 7.3 Hz, 1H), 7.34 (d, J = 5.8 Hz, 1H),
.10 (d, J = 5.8 Hz, 1H), 7.07 (dd, J = 7.3, 7.3 Hz, 1H), 6.97 (dd, J =
.3, 7.3 Hz, 1H), 6.79 (dd, J = 7.8 Hz, 1H), 6.75 (s, 1H), 6.69 (d, J =
3
/ 0.5%AcOH.
1
7.34 (d, J = 7.2 Hz, 1H), 7.10 (d, J = 7.2 Hz, 1H), 7.08 (dd, J = 7.7,
7
6
.7 Hz, 1H), 6.98 (dd, J = 7.5, 7.5 Hz, 1H), 6.80 (dd, J = 8.2 Hz, 1H),
.76 (s, 1H), 6.70 (d, J = 8.2 Hz, 1H), 6.50 (d, J = 15.4 Hz, 1H), 6.39
6
7
5
7
7
(
3
ddd, J = 15.4, 7.2, 7.2 Hz, 1H), 5.55 (s, 1H), 5.03 (d, J = 6.4 Hz, 1H),
.69–3.66 (m, 1H), 3.50 (dd, J = 16.5, 7.1 Hz,1H), 3.39–3.35 (m,1H),
3.33 & 2.83 (AB quartet, J = 13.6 Hz, 2H), 3.26 (dd, J = 16.4, 6.8Hz,
1
2
2
H), 3.17 (d, J = 15.3 Hz,1H), 3.13–3.04 (m, 2H), 3.02–2.94 (m, 1H),
.83 (dd, J = 15.3, 6.3 Hz, 1H), 2.58 (dd, J = 2.5, 2.5 Hz, 1H), 2.54–
.52 (m,1H), 2.34 (ddd, J = 13.8, 10.7, 3.1 Hz, 1H), 2.25 & 2.09 (AB
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
7.8 Hz, 1H), 6.47 (d, J = 15.6 Hz, 1H), 6.38 (ddd, J = 15.6, 6.7, 6.7
Hz, 1H), 5.56 (s, 1H), 5.00 (d, J = 5.6 Hz, 1H), 3.62–3.57 (m, 1H),
3
.49 (dd, J = 16.7, 7.0 Hz, 1H), 3.38–3.32 (m, 1H), 3.31 & 2.86 (AB
quartet, J = 16.3 Hz, 2H), 1.92 (ddd, J = 6.8, 6.8, 2.5 Hz, 2H), 1.45–
1.39 (m,1H), 1.45–1.39 (m,1H), 1.38–1.31 (m, 4H), 1.24–1.18 (m,
1H), 1.19–1.13 (m, 1H); C NMR (DMSO-d
1
1
1
quartet, J = 14.0 Hz, 2H), 3.26 (dd, J = 16.7, 6.8Hz, 1H), 3.21 (d, J =
1
1
5.1 Hz, 1H), 2.99–2.93 (m, 1H), 2.99–2.93 (m, 1H), 2.80 (dd, J =
5.1, 5.6 Hz, 1H), 2.58 (dd, J = 2.0, 2.0 Hz, 1H), 2.48–2.45 (m, 1H),
1
3
6
, 151 MHz): δ 171.3,
70.7, 169.8, 152.5, 137.3, 137.1, 136.3, 131.1, 129.9, 129.8, 129.3,
29.1, 128.8, 128.7, 128.2, 126.5, 125.9, 125.3, 123.4, 120.6, 118.2,
16.9, 114.1, 110.8, 105.9, 83.5, 70.7, 57.2, 52.0, 49.8, 48.8, 44.5,
2.32 & 2.09 (AB quartet, J = 16.0 Hz, 2H), 2.01–1.88 (m, 2H), 1.92
ddd, J = 6.4, 6.4, 2.0 Hz, 2H), 1.61–1.51 (m, 2H), 1.45–1.41 (m, 1H),
.36–1.32 (m, 1H), 1.34–1.30 (m, 1H), 1.17 (ddd, J = 13.1, 13.1, 4.1
(
1
13
39.5, 39.4, 38.0, 34.7, 34.6, 34.2, 31.5, 28.3, 24.8, 22.4, 17.9; HRMS
Hz, 1H); C NMR (DMSO-d
69.4, 152.6, 137.7, 137.4, 136.1, 131.0, 129.9, 129.7, 129.5, 129.2,
128.9, 128.9, 128.2, 126.6, 126.0, 125.2, 123.6, 120.5, 118.3, 117.1,
6
, 151 MHz): δ 173.6, 171.6, 171.1,
+
(QE Orbitrap) m/z: [M+H] calc’d for C43
24.3592.
E)-3-(3-((S)-2-(diethoxymethyl)-2-(2-(((R)-1-(((2S,3R)-3-
hydroxy-1-((4-hydroxyphenethyl)amino)-1-oxobutan-2-yl)amino)-
-(3-methoxyphenyl)-1-oxopropan-2-yl)amino)-2-oxoethyl)hept-6-
46 7 4
H N O 724.3606; found
1
7
(
1
3
14.2, 110.8, 105.7, 83.6, 70.8, 57.3, 52.7, 48.8, 44.2, 39.6, 39.4,
8.3, 34.4, 34.2, 31.5, 31.4, 26.9, 24.5, 22.5, 18.1; HRMS (QE Or-
+
3
bitrap) m/z: [M+H] calc’d for C43
H
45
N
4
O
6
713.3334; found 713.3321.
yn-1-yl)phenyl)allyl isobutyl carbonate (S29)
Synthesized according to Procedure A beginning with 0.17 mmol
of (+)-5. Carried forward without purification.
1
5
11b
3
1
3
-((1 S,1 R,1
S,13S,E)-1 ,12,15-trioxo-1 -(pent-4-yn-1-yl)-
2
3 5 6 11 11b
1
1
,1 ,1 ,1 ,1 ,1
-hexahydro-1 H-7-oxa-11,14-diaza-1(1,5)-
indolizino[8,7-b]indola-3(1,3),8(1,4)-
dibenzenacyclopentadecaphan-4-en-13-yl)propanoic acid (40)
Synthesized according to Procedure D using 50 mg of crude S26.
Pictet-Spengler Products S30a & b
Synthesized according to Procedure B using crude material from
the previous reaction. Carried forward without purification.
1
5
10b
6
8
Purified by preparative TLC: 3% MeOH / 96.5%CHCl
White solid. 8 mg [11 µmol, 19% yield over three steps]. H NMR
CDCl , 500 MHz): δ11.06 (s, 1H), 8.23 (d, J = 6.9 Hz, 1H), 7.52 (d,
3
/ 0.5%AcOH.
(1 S,1 R,1 S,12S,E)-7 -hydroxy-12-((R)-1-hydroxyethyl)-1 -
1
1
1
2
3
5
6
10b
methoxy-1 -(pent-4-yn-1-yl)-1 ,1 ,1 ,1 ,1 ,1 -hexahydro-10,13-
diaza-1(1,5)-pyrrolo[2,1-a]isoquinolina-3,7(1,3)-
dibenzenacyclotetradecaphan-4-ene-1 ,11,14-trione (42)
(
3
3
J = 8.0 Hz, 1H), 7.45–7.42 (m, 1H), 7.43 (d, J = 7.9 Hz, 1H), 7.38 (d,
J = 8.0 Hz, 1H), 7.35 (d, J = 7.8 Hz, 1H), 7.33 (d, J = 2.3 Hz, 1H),
Synthesized according to Procedure C using 26 mg of crude S30.
7
2
.14 (d, J = 8.6 Hz, 1H), 7.10 (d, J = 7.6 Hz, 1H), 7.07 (d, J = 8.4 Hz,
H), 7.00 (dd, J = 7.7, 7.7 Hz, 1H), 6.83 (d, J = 8.3 Hz, 2H), 6.71 (d,
Purified by preparative HPLC – see SI for conditions. White solid. 16
mg [24 µmol, 49% yield over three steps]. H NMR (DMSO-d
1
6
, 600
J = 15.9 Hz, 1H), 6.33 (ddd, J = 15.9, 6.6, 4.8 Hz, 1H), 5.25 (s, 1H),
.06 (d, J = 6.6 Hz, 1H), 4.90 (dd, J = 15.0, 7.0 Hz, 1H), 4.83 (dd, J =
MHz): δ 7.51 (d, J = 7.6 Hz, 1H), 7.48 (dd, J = 5.8, 5.8 Hz, 1H), 7.42
s, 1H), 7.36-7.33 (m, 1H), 7.36-7.33 (m, 1H), 7.36-7.33 (m, 1H),
.10 (d, J = 4.7 Hz, 1H), 6.85 (s, 1H), 6.82 (d, J = 8.2 Hz, 1H), 6.80
d, J = 8.2 Hz, 1H), 6.70 (d, J = 8.2 Hz, 1H), 6.68 (s, 1H), 6.51 (d, J =
15.7 Hz, 1H), 6.38 (ddd, J = 15.6, 7.2 Hz, 1H), 5.37 (s, 1H), 5.04 (d, J
6.4 Hz, 1H), 3.77-3.71 (m, 1H), 3.73-3.71 (m, 1H), 3.72 (s, 3 H),
5
(
7
(
15.0, 4.7 Hz, 1H), 4.11 (dd, J = 5.3, 5.3 Hz, 1H), 3.70–3.65 (m, 1H),
3.13 (d, J = 15.3 Hz, 1H), 3.01–2.96 (m, 1H), 2.90 (dd, J = 15.0, 7.0
Hz, 1H), 2.84 (d, J = 13.9 Hz, 1H), 2.74–2.67 (m, 1 H), 2.45–2.36 (m,
2
H), 2.23–2.16 (m, 1H), 2.16 (d, J = 16.3 Hz, 1H), 2.08 (d, J = 16.2
Hz, 1H), 2.02–1.95 (m, 1H), 1.93 (ddd, J = 6.9, 6.9, 2.0 Hz, 2H),
.80–1.73 (m, 1H), 1.70–1.62 (m, 1H), 1.43–1.23 (m, 6H), 1.12–1.06
=
3
1
3
.48 (dd, J = 16.4, 7.0 Hz, 1H), 3.36 (dddd, J = 6.3, 6.3, 6.3, 6.3 Hz,
H), 3.31 (dd, J = 16.5, 6.9 Hz, 1H), 3.16 (d, J = 13.9 Hz, 1H), 3.06-
.02 (m, 1H), 3.06 (d, J = 15.9 Hz, 1H), 2.90 (dd, J = 16.1, 6.1Hz,
1
1
3
(m, 1H); C NMR (CDCl
3
, 126 MHz): δ173.8, 171.4, 171.2, 171.0,
1
1
1
3
55.8, 137.8, 136.7, 135.9, 132.9, 131.7, 131.5, 130.7, 130.04,
1H), 2.85 (d, J = 14.0 Hz, 1H), 2.64 (t, J = 2.1 Hz, 1H), 2.54-2.48 (m,
H), 2.45-2.41 (m, 2H), 2.39 (d, J = 16.8 Hz, 1H), 2.09 (d, J = 16.6
Hz, 1H), 1.92-1.90 (m, 2H), 1.39-1.29 (m,1H), 1.31-1.29 (m, 1H),
29.97, 128.8, 126.3, 126.2, 123.0, 121.1, 118.8, 117.3, 115.5, 111.5,
06.0, 84.2, 71.3, 67.6, 57.7, 52.6, 48.9, 45.2, 40.4, 38.0, 34.4, 34.0,
1
+
1
3
2.4, 30.5, 26.6, 26.1, 22.6, 18.2; HRMS (QE Orbitrap) m/z: [M+H]
1
.19-1.15 (m, 1H), 1.19-1.15 (m, 1H), 0.86 (d, J = 6.1 Hz, 3H);
C
45 4 6
calc’d for C43H N O 713.3334; found 713.3324.
(E)-3-(3-((S)-2-(2-(((R)-1-(((S)-5-azido-1-((4-
hydroxyphenethyl)amino)-1-oxopentan-2-yl)amino)-3-(1H-indol-
6
NMR (DMSO-d , 151 MHz): δ 171.1, 169.5, 169.4, 157.1, 152.6,
1
126.7, 126.5, 125.6, 125.0, 123.9, 114.3, 113.9, 112.3, 83.8, 71.0,
6
3
37.5, 137.5, 133.8, 131.5, 130.1, 129.6, 129.2, 128.8, 128.3, 128.2,
3
-yl)-1-oxopropan-2-yl)amino)-2-oxoethyl)-2-
6.1, 58.7, 58.2, 55.7, 48.5, 45.3, 40.8, 40.0, 38.8, 34.8, 34.3, 32.3,
+
(
(
diethoxymethyl)hept-6-yn-1-yl)phenyl)allyl isobutyl carbonate
S27)
1.4, 22.3, 19.7, 18.0; HRMS (QE Orbitrap) m/z: [M+H] calc’d for
C
41
H
46
N
3
O
6
676.3381; found 676.3368.
Synthesized according to Procedure A beginning with 0.19 mmol
(
E)-3-(3-((S)-2-(2-(((R)-3-(5-bromo-1H-indol-3-yl)-1-(((S)-1-((4-
of (+)-5. Carried forward without purification.
E)-3-(3-(((1S,5R,11bS)-5-(((S)-5-azido-1-((4-
hydroxyphenethyl)amino)-1-oxopentan-2-yl)carbamoyl)-3-oxo-1-
pent-4-yn-1-yl)-2,3,5,6,11,11b-hexahydro-1H-indolizino[8,7-
hydroxyphenethyl)amino)-3-(1H-imidazol-4-yl)-1-oxopropan-2-
yl)amino)-1-oxopropan-2-yl)amino)-2-oxoethyl)-2-
(
(
(
diethoxymethyl)hept-6-yn-1-yl)phenyl)allyl isobutyl carbonate
S31)
(
b]indol-1-yl)methyl)phenyl)allyl isobutyl carbonate (S28)
Synthesized according to Procedure B using crude material from
the previous reaction. Carried forward without purification.
Synthesized according to Procedure A beginning with 0.17 mmol
of (+)-5. Carried forward without purification.
E)-3-(3-(((1S,5R,11bS)-8-bromo-5-(((S)-1-((4-
hydroxyphenethyl)amino)-3-(1H-imidazol-4-yl)-1-oxopropan-2-
yl)carbamoyl)-3-oxo-1-(pent-4-yn-1-yl)-2,3,5,6,11,11b-hexahydro-
1
(
1
5
11b
6
1
(
1 S,1 R,1 S,12S,E)-12-(3-azidopropyl)-7 -hydroxy-1 -(pent-4-
2 3 5 6 11 11b 1
yn-1-yl)-1 ,1 ,1 ,1 ,1 ,1 -hexahydro-1 H-10,13-diaza-1(1,5)-
indolizino[8,7-b]indola-3,7(1,3)-dibenzenacyclotetradecaphan-4-
ene-1 ,11,14-trione (41)
H-indolizino[8,7-b]indol-1-yl)methyl)phenyl)allyl isobutyl car-
3
bonate (S32)
ACS Paragon Plus Environment

Page 15 of 20
The Journal of Organic Chemistry
1
5
11b
6
Synthesized according to a modified Procedure B using crude ma-
terial from the previous reaction: 10 vol% of conc. H PO was added
(1 S,1 R,1 S,12R,E)-12-((tert-butyldisulfaneyl)methyl)-7 -
2 3 5 6 11 11b 1
1
2
3
4
5
6
7
8
9
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
2
2
2
3
3
3
3
3
3
3
3
3
3
4
4
4
4
4
4
4
4
4
4
5
5
5
5
5
5
5
5
5
5
6
3
4
hydroxy-11-(pent-4-yn-1-yl)-1 ,1 ,1 ,1 ,1 ,1 -hexahydro-1 H-
10,13-diaza-1(1,5)-indolizino[8,7-b]indola-3,7(1,3)-
dibenzenacyclotetradecaphan-4-ene-1 ,11,14-trione (45a)
Synthesized according to Procedure C using crude material from
previous reaction. Purified by preparative HPLC – see SI for condi-
tions. Yellow solid. 32 mg [41 µmol, 24% yield over three steps]. H
to the aqueous acetic acid solution. Carried forward without purifica-
tion.1
3
5
11b
8
(
1 S,1 R,1 S,12S,E)-12-((1H-imidazol-4-yl)methyl)-1 -bromo-
6
1
2
3
5
6
11 11b
1
7
-hydroxy-1 -(pent-4-yn-1-yl)-1 ,1 ,1 ,1 ,1 ,1 -hexahydro-1 H-
1
10,13-diaza-1(1,5)-indolizino[8,7-b]indola-3,7(1,3)-
3
dibenzenacyclotetradecaphan-4-ene-1 ,11,14-trione (43)
6
NMR (DMSO-d , 600 MHz): δ 10.96 (s, 1H), 9.10 (s, 1H), 7.87 (d, J
Synthesized according to Procedure C using 43 mg of crude S32.
= 7.3 Hz, 1H), 7.64 (dd, J = 5.7, 5.7 Hz, 1H), 7.45 (s, 1H), 7.38 (d, J
= 8.2 Hz, 1H), 7.35-7.33 (m, 1H), 7.35-7.33 (m, 1H), 7.35 (d, J = 8.3
Hz, 1H), 7.08 (dd, J = 8.2, 8.2 Hz, 1H), 7.08 (d, J = 7.0 Hz, 1H), 6.78
(d, J = 8.3 Hz, 1H), 6.77 (s, 1H), 6.68 (d, J = 8.0 Hz, 1H), 6.46 (d, J =
15.7 Hz, 1H), 6.40 (ddd, J = 15.6, 6.6, 6.6 Hz, 1H), 5.59 (s, 1H), 5.04
(d, J = 6.4 Hz, 1H), 3.96 (ddd, J = 10.8, 7.3, 3.8 Hz, 1H), 3.51 (dd, J
= 16.2, 6.7 Hz,1H), 3.36 & 2.81 (AB quartet, J = 14.3 Hz, 2H), 3.29
(d, J = 15.1 Hz,1H), 3.28-3.25 (m,1H), 3.24 (dd, J = 16.4, 5.8 Hz,
1H), 3.12-3.07 (m, 1H), 2.81 (dd, J = 15.0, 6.2 Hz, 1H), 2.73 (dd, J =
12.8, 3.6 Hz,1H), 2.63 (dd, J = 12.9, 10.9 Hz, 1H), 2.57 (t, J = 2.3 Hz,
1H), 2.56-2.52 (m,1H), 2.34 (ddd, J = 14.0, 11.0, 3.0 Hz, 1H), 2.24 &
2.08 (AB quartet, J = 16.2 Hz, 2H), 1.92-1.91 (m, 2H), 1.44-1.40
(m,1H), 1.36-1.31 (m,1H), 1.35-1.32 (m, 1H), 1.20-1.16 (m, 1H), 1.19
Purified by preparative HPLC – see SI for conditions. White solid.
1
TFA salt: 30 mg [33 µmol, 61% yield over three steps]. H NMR
(DMSO-d
6
, 600 MHz): δ 11.20 (s, 1H), 9.20 (s, 1H), 8.81 (s, 1H),
8
1
7
.11 (d, J = 4.9 Hz, 1H), 7.46 (s, 1H), 7.44 (s, 1H), 7.42–7.40 (m,
H), 7.37–7.35 (m, 1H), 7.36–7.35 (m, 1H), 7.34 (d, J = 8.3 Hz, 1H),
.20 (d, J = 8.2 Hz, 1H), 7.15 (s, 1H), 7.10 (d, J = 4.7 Hz, 1H), 6.81
d, J = 7.2 Hz, 1H), 6.77 (s, 1H), 6.71 (d, J = 7.3 Hz, 1H), 6.49 (d, J =
15.7 Hz, 1H), 6.38 (ddd, J = 15.2, 6.5, 6.5 Hz, 1H), 5.49 (s, 1H), 5.01
(d, J = 4.5 Hz, 1H), 4.05–4.02 (m, 1H), 3.47 (dd, J = 16.1, 5.9 Hz,
H), 3.38–3.34 (m, 1H), 3.32 (dd, J = 16.4, 6.1 Hz, 1H), 3.30 & 2.84
AB quartet, J = 12.2 Hz, 2H), 3.07–3.03 (m, 1H), 2.95 (d, J = 15.3
Hz, 1H), 2.87–2.83 (m, 1), 2.78–2.75 (m, 1H), 2.77 (dd, J = 15.3, 4.6
Hz, 1H), 2.56 (br s, 1H), 2.50–2.48 (m, 1H), 2.39–2.36 (m, 1H), 2.26
& 2.10 (AB quartet, J = 16.4 Hz, 2H), 1.91 (very broad singlet, 2H),
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
(
1
(
1
3
6
(s, 9H); C NMR (DMSO-d , 151 MHz): δ 171.2, 169.8, 169.2,
152.8, 137.5, 137.4, 136.5, 131.0, 130.0, 129.7, 129.4, 129.3, 129.1,
128.9, 128.1, 126.9, 126.2, 125.6, 123.6, 120.6, 118.2, 117.3, 114.1,
110.9, 106.2, 83.8, 70.9, 57.3, 52.6, 48.9, 47.2, 44.3, 41.9, 39.7, 39.5,
1
(
1
1
.40–1.37 (m, 1H), 1.32–1.29 (m, 1H), 1.32–1.29 (m, 1H), 1.17–1.13
13
m, 1H); C NMR (DMSO-d
6
, 151 MHz): δ 171.4, 170.1, 169.8,
52.8, 137.5, 137.4, 135.1, 133.4, 131.5, 131.3, 130.2, 129.3, 129.2,
29.1, 128.9, 128.7, 128.3, 127.9, 126.7, 125.7, 123.8, 123.3, 119.4,
38.1, 34.8, 34.2, 31.7, 29.1, 24.2, 22.6, 18.1; HRMS (DART-
+
Orbitrap) m/z: [M+H] calc’d for C45
H
51
N
4
O
4
S
2
775.3346; found
116.2, 114.4, 113.1, 111.1, 105.7, 83.8, 71.0, 57.3, 51.8, 48.5, 44.5,
775.3365.
1
5
11b
3
1
39.8, 39.4, 38.0, 34.6, 34.1, 31.4, 26.4, 24.5, 22.4, 18.0; HRMS (QE
(1 S,1 R,1 S,10R,E)-N-(4-hydroxyphenethyl)-1 ,12-dioxo-1 -
(pent-4-yn-1-yl)-1 ,1 ,1 ,1 ,1 ,11 -hexahydro-1 H-7,8-dithia-11-
aza-1(1,5)-indolizino[8,7-b]indola-3(1,3)-
+
2 3 5 6 11 1b 1
Orbitrap) m/z: [M+H] calc’d for C44
H
44BrN
6
O
4
799.2602; found
7
99.2595.
1 5
11
8
(
1 S,1 R,1 bS,13S,E)-13-((1H-imidazol-4-yl)methyl)-1 -bromo-
benzenacyclododecaphan-4-ene-10-carboxamide (45b)
Purified by preparative HPLC from the above reaction – see SI for
1
2
3
5
6
11 11b
1
1
-(pent-4-yn-1-yl)-1 ,1 ,1 ,1 ,1 ,1 -hexahydro-1 H-7-oxa-11,14-
diaza-1(1,5)-indolizino[8,7-b]indola-3(1,3),8(1,4)-
conditions. Pale yellow solid. 33 [46 µmol, 27% yield over three
3
1
dibenzenacyclopentadecaphan-4-ene-1 ,12,15-trione (44)
steps]. H NMR (DMSO-d
6
, 600 MHz): δ 10.88 (s, 1H), 9.14 (br s,
Synthesized according to Procedure E using 50 mg of crude Pictet-
Spengler product (S32). Purified by preparative HPLC using HCOOH
1H), 7.93 (t, J = 5.6 Hz, 1H), 7.69 (d, J = 7.0 Hz, 1H), 7.65 (s, 1H),
7.45 (d, J = 7.7 Hz, 1H), 7.42 (d, J = 8.0 Hz, 1H), 7.38 (dd, J = 7.6,
7.6 Hz, 1H), 7.21 (d, J = 7.5 Hz, 1H), 7.16 (d, J = 7.6 Hz, 1H), 7.10
(dd, J = 7.5, 7.5 Hz, 1H), 7.01 (dd, J = 7.5, 7.5 Hz, 1H), 6.91 (d, J =
8.2 Hz, 2H), 6.65 (d, J = 8.2 Hz, 2H), 6.58 (d, J = 15.4 Hz, 1H), 6.24
(ddd, J = 15.5, 9.1, 6.2 Hz, 1H), 5.25 (s, 1H), 4.93 (d, J = 5.8 Hz, 1H),
4.02 (ddd, J = 10.7, 6.8, 3.9 Hz, 1H), 3.62 (dd, J = 13.9, 9.2 Hz,1H),
3.46 (dd, J = 14.0, 6.2 Hz, 1H), 3.39 (d, J = 15.5 Hz,1H), 3.34 & 2.86
(AB quartet, J = 13.8 Hz, 2H), 3.14-3.03 (m, 2H), 2.76-2.74 (m, 1H),
2.73 (dd, J = 11.8, 11.8 Hz,1H), 2.57 (t, J = 2.3 Hz, 1H), 2.47 (t, J =
7.4 Hz, 2H), 2.37 (dd, J = 12.3, 3.6 Hz, 1H), 2.31 & 2.12 (AB quartet,
instead of TFA – see SI for conditions. White solid. 15 mg [19 µmol,
1
30% yield over three steps]. H NMR (DMSO-d
6
, 600 MHz): δ 11.28
(
s, 1H), 8.85 (s,1H), 8.45 (d, J = 7.2 Hz, 1H), 7.50 (d, J = 7.7 Hz, 1H),
7
7
8
.43 (d, J = 1.5 Hz, 1H), 7.38 (d, J = 8.5 Hz, 1H), 7.34-7.32 (m, 1H),
.34 (s,1H), 7.34 (dd, J = 7.5, 7.5 Hz, 1H), 7.31 (s, 1H), 7.23 (dd, J =
.3 Hz, 1H), 7.15 (d, J = 7.4 Hz, 1H), 7.04 (d, J = 8.3 Hz, 1H), 6.84
(d, J = 8.3 Hz, 1H), 6.66 (d, J = 16.1 Hz, 1H), 6.33 (ddd, J = 16.1,
5.7, 5.7 Hz, 1H), 5.15 (s, 1H), 5.06 (dd, J = 5.6, 1.4 Hz, 1H), 4.90 (dd,
J = 15.0, 6.1 Hz, 1H), 4.85 (dd, J = 15.0, 4.4 Hz, 1H), 4.09 (ddd, J =
.4, 7.4, 7.4 Hz, 1H), 3.23-3.19 (m, 1H), 3.22 & 2.85 (AB quartet, J =
3.8 Hz, 2H), 3.09-3.06 (m, 1H), 2.99 (d, J = 14.9, 5.8 Hz, 1H), 2.87-
.85 (m, 1H), 2.84-2.83 (m, 2H), 2.70-2.67 (m, 1H), 2.58 (dd, J = 2.1,
7
1
2
J = 16.4 Hz, 2H), 1.94-1.92 (m, 2H), 1.51-1.45 (m, 1H), 1.43-1.39 (m,
13
6
1H), 1.35-1.32 (m, 1H), 1.35-1.32 (m, 1H); C NMR (DMSO-d , 151
MHz): δ 171.4, 168.1, 168.0, 155.4, 137.3, 136.6, 136.6, 131.5, 130.5,
129.5, 129.2, 129.1, 128.3, 127.1, 127.0, 124.1, 124.0, 120.8, 118.3,
117.6, 114.7, 111.0, 106.8, 83.8, 70.9, 55.9, 52.8, 49.8, 43.4, 43.2,
2.1 Hz, 1H), 2.41-2.37 (m, 1H), 2.18 & 2.10 (AB quartet, J = 16.4 Hz,
H), 1.92 (ddd, J = 6.7, 6.7, 2.0 Hz, 2H), 1.40-1.30 (m, 2H), 1.28-1.25
2
(
1
1
1
3
m, 1H), 1.06 (ddd, J = 12.7, 12.7, 3.7 Hz, 1H); C NMR (DMSO-d
6
,
40.7, 40.5, 39.9, 38.2, 35.1, 33.8, 22.8, 22.3, 18.1; HRMS (QE Or-
+
51 MHz): δ 171.1, 171.0, 170.0, 155.6, 137.4, 135.8, 135.1, 133.3,
32.5, 131.5, 131.3, 131.2, 130.0, 130.0, 129.1, 128.6, 127.9, 125.9,
123.3, 123.1, 119.3, 117.0, 115.4, 113.2, 111.3, 105.5, 83.9, 71.0,
bitrap) m/z: [M+H] calc’d for C41
H
43
N
4
O
4
S
2
719.2720; found
719.27119.
5
11b
6
3
3-((1 S,1 R,1 S,9S,12S,E)-9-carbamoyl-7 -hydroxy-1 ,11,14-
1
67.3, 57.2, 51.9, 48.3, 45.0, 40.1, 38.5, 37.8, 34.1, 32.4, 26.1, 25.3,
trioxo-1 -(3-(1-((2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-
+
2
C
2.4, 18.0; HRMS (QE Orbitrap) m/z: [M+H] calc’d for
(hydroxymethyl)tetrahydro-2H-pyran-2-yl)-1H-1,2,3-triazol-4-
2
3
5
6
11 11b
1
44
H
44BrN
6
O
4
799.2602; found 799.2576.
yl)propyl)-1 ,1 ,1 ,1 ,1 ,1 -hexahydro-1 H-10,13-diaza-1(1,5)-
indolizino[8,7-b]indola-3,7(1,3)-dibenzenacyclotetradecaphan-4-
en-12-yl)propanoic acid (46)
(
E)-3-(3-((S)-2-(2-(((R)-1-(((R)-3-(tert-butyldisulfaneyl)-1-((4-
hydroxyphenethyl)amino)-1-oxopropan-2-yl)amino)-3-(1H-indol-
3-yl)-1-oxopropan-2-yl)amino)-2-oxoethyl)-2-
diethoxymethyl)hept-6-yn-1-yl)phenyl)allyl isobutyl carbonate
S33)
Synthesized according to Procedure A beginning with 0.17 mmol
of (+)-5. Carried forward without purification.
(E)-3-(3-(((1S,5R,11bS)-5-(((R)-3-(tert-butyldisulfaneyl)-1-((4-
hydroxyphenethyl)amino)-1-oxopropan-2-yl)carbamoyl)-3-oxo-1-
Synthesized according to Procedure F with 13 µmol of 36. Puri-
(
(
fied by preparative HPLC – see SI for conditions. White solid. 4 mg
1
[4.2 µmol, 32% yield]. H NMR (DMSO-d
6
, 500 MHz): δ 7.73 (s,
1H), 7.48 (br s, 1H), 7.38 (d, J = 7.9 Hz, 1H), 7.37–7.31 (m, 2H), 7.09
(d, J = 6.9 Hz, 1H), 7.08 (d, 7.4 Hz, 1H), 6.99 (d, J = 7.8 Hz, 1H),
6.97 (d, J = 4.9 Hz, 1H), 6.82 (d, J = 8.1 Hz, 1H), 6.77 (br s, 1H),
6.66 (J = 8.1 Hz, 1H), 6.52 (d, J = 15.4 Hz, 1H), 6.37 (ddd, J = 15.3,
7.2, 7.2 Hz, 1H), 5.67 (s, 1H), 5.34 (d, J = 9.4 Hz, 1H), 5.00 (d, J =
6.6 Hz, 1H), 4.14 (ddd, J = 7.9, 7.9, 2.7 Hz, 1H), 3.76–3.73 (m, 1H),
3.70–3.66 (m, 2H), 3.53–3.38 (m, 3H) 3.23–3.18 (m, 1H), 2.95–2.86
(m, 1H), 2.63–2.60 (m, 1H), 2.36 (dd, J = 7.6, 7.6 Hz, 2H), 2.22 (d, J
(
pent-4-yn-1-yl)-2,3,5,6,11,11b-hexahydro-1H-indolizino[8,7-
b]indol-1-yl)methyl)phenyl)allyl isobutyl carbonate (S34)
Synthesized according to Procedure B using crude material from
previous reaction. Carried forward without purification.
ACS Paragon Plus Environment

The Journal of Organic Chemistry
Page 16 of 20
=
16.4 Hz, 1H), 2.15 (d, J = 16.0 Hz, 1H), 2.08–2.03 (m, 1H), 1.99–
1.94 (m, 1H), 1.78–1.73 (m, 1H), 1.64–1.59 (m, 2H), 1.55–1.50 (m,
H), 1.38–1.31 (m, 1H), 1.27–1.21 (m, 1H); HRMS (QE Orbitrap)
1H), 5.05 (dd, J = 6.3, 1.3 Hz, 1H), 3.89 (dd, J = 12.3, 2.0 Hz, 1H),
1
2
3
4
5
6
7
8
9
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
2
2
2
3
3
3
3
3
3
3
3
3
3
4
4
4
4
4
4
4
4
4
4
5
5
5
5
5
5
5
5
5
5
6
3.86 (dd, J = 9.1, 9.1 Hz, 1H), 3.78–3.71 (m, 3H), 3.65–3.59 (m, 1H),
3.57 (ddd, J = 4.8, 4.8, 1.7 Hz, 1H), 3.56–3.50 (m, 2H), 3.47–3.44 (m,
1H), 3.34 (dd, J = 15.6, 6.0 Hz, 1H), 3.28 (dd, J = 5.3, 5.3 Hz, 1H),
3.27–3.21 (m, 2H), 2.95 (dd, J = 16.3, 6.3 Hz, 1H), 2.92 (d, J = 14.2
Hz, 1H), 2.68 (d, J = 16.7 Hz, 1H), 2.62 (ddd, J = 14.0, 5.2, 5.2 Hz,
1H), 2.58–2.50 (m, 2H), 2.48–2.42 (m, 1H), 2.21 (d, J = 16.6 Hz, 1H),
1.63–1.54 (m, 1H), 1.49–1.40 (m, 1H), 1.36, (ddd, J = 13.0, 13.0, 3.8
1
+
m/z: [M+H] calc’d for C50H N O12 961.4091; found 961.4087.
57 8
1
5
11b
6
(
1 S,1 R,1 S,9S,12S,E)-12-(3-amino-3-oxopropyl)-7 -hydroxy-
,11,14-trioxo-1 -(3-(1-((2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-
3
1
1
(hydroxymethyl)tetrahydro-2H-pyran-2-yl)-1H-1,2,3-triazol-4-
2
3
6
11 11b
1
yl)propyl)-1 ,1 ,15,1 ,1 ,1 -hexahydro-1 H-10,13-diaza-1(1,5)-
indolizino[8,7-b]indola-3,7(1,3)-dibenzenacyclotetradecaphan-4-
ene-9-carboxamide (S10)
Hz, 1H), 1.24 (ddd, J = 13.3, 13.3 4.5 Hz, 1H), 0.82 (d, J = 6.4 Hz,
13
3
1H); C NMR (CD OD, 126 MHz): δ 176.0, 171.6, 171.6, 159.9,
Synthesized according to Procedure F with 13 µmol of 37. Puri-
154.4, 148.5, 140.1, 138.9, 135.3, 132.6, 131.3, 131.2, 131.0, 130.6,
130.1, 129.8, 128.8, 128.6, 128.1, 125.6, 125.4, 122.3, 115.5, 115.4,
114.6, 89.5, 81.1, 78.5, 73.9, 70.9, 68.2, 62.5, 61.3, 60.4, 55.8, 51.4,
fied by preparative HPLC – see SI for conditions. White solid. 5 mg
1
[
7
(
7
5.2 µmol, 40% yield]. H NMR (CD
3
OD, 500 MHz): δ 7.85 (s, 1H),
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
.47 (s, 1H), 7.49 (d, J = 8.1 Hz, 1H), 7.41 (d, J = 7.7 Hz, 1H), 7.15
d, J = 7.4 Hz, 1H), 7.10 (dd, J = 7.5, 7.5 Hz, 1H), 7.04 (dd, J = 7.5,
.5 Hz, 1H), 6.79 (d, J = 8.1 Hz, 1H), 6.62 (d, J = 8.3 Hz, 1H), 6.60–
6.55 (m, 1H), 6.53 (d, J = 16.0 Hz, 1H), 5.77 (s, 1H), 5.32 (d, J = 9.2
Hz, 1H), 5.19 (d, J = 6.0 Hz, 1H), 4.17 (d, J = 10.7 Hz, 1H), 3.89 (d, J
12.1 Hz, 1H), 3.86–3.83 (m, 1H), 3.83 (dd, J = 9.3, 9.3 Hz, 1H),
.72 (d, J = 12.2, 5.2 Hz, 1H), 3.56–3.47 (m, 6H), 3.13 (d, 13.8 Hz,
H), 2.95 (d, J = 16.4, 1H), 2.89 (dd, J = 15.4, 7.6 Hz, 1H), 2.75 (d, J
48.2, 42.7, 41.3, 40.6, 36.7, 35.8, 33.4, 32.4, 26.7, 24.4, 19.7; HRMS
+
(QE Orbitrap) m/z: [M+H] calc’d for C47
H
57
N
6
O
11 881.4080; found
881.40815.
5
11b
6
3
1
3-((1 S,1 R,1 S,12S,E)-7 -hydroxy-1 ,11,14-trioxo-1 -(3-(1-
((2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-
2
3
5
6
11 11b
=
3
1
2H-pyran-2-yl)-1H-1,2,3-triazol-4-yl)propyl)-1 ,1 ,1 ,1 ,1 ,1
-
1
hexahydro-1 H-10,13-diaza-1(1,5)-indolizino[8,7-b]indola-
3,7(1,3)-dibenzenacyclotetradecaphan-4-en-12-yl)propanoic acid
(S13)
Synthesized according to Procedure F beginning with 15 µmol of
=
13.3 Hz, 1H), 2.60–2.50 (m, 2H), 2.47 (d, J = 16.4 Hz, 1H), 2.17–
2.08 (m, 2H), 1.81–1.74 (m, 1H), 1.72–1.63 (m, 3H), 1.42–1.32 (m, 3
1
3
H), 1.32–1.27 (m, 3H), 1.13–1.09 (m, 1H); C NMR (CD
3
OD, 126
39. Purified by preparative HPLC – see SI for conditions. White
1
MHz): δ 177.8, 176.7, 175.4, 174.2, 173.2, 154.6, 148.5, 140.0, 139.5,
37.9, 132.5, 131.0, 130.8, 130.5, 130.0, 129.9, 129.8, 128.9, 128.3,
27.5, 127.1, 127.0, 123.5, 122.4, 121.0, 119.5, 115.9, 112.1, 108.4,
solid. 4 mg [4.4 µmol, 29% yield]. H NMR (DMSO-d
6
, 500 MHz): δ
1
1
8.19 (d, J = 5.3 Hz, 1H), 7.47 (s, 1H), 7.46 (d, J = 5.6 Hz, 1H), 7.36
(ddd, J = 8.0, 1.2, 1.2 Hz, 1H), 7.34 (ddd, J = 8.1, 0.8, 0.8 Hz, 1H),
7.32 (dd, J = 7.6, 7.6 Hz, 1H), 7.31 (s, 1H), 7.15 (ddd, J = 8.1, 7.2, 0.9
Hz, 1H), 7.05 (ddd, J = 7.8, 7.0, 0.8 Hz, 1H), 7.03 (ddd, J = 7. 1, 1.2,
1.2 Hz, 1H), 6.91 (d, J = 1.8 Hz, 1H), 6.73 (dd, J = 8.2, 2.1 Hz, 1H),
6.60 (d, J = 8.1 Hz, 1H), 6.51 (ddd, 15.8, 6.1, 6.1 Hz, 1H), 6.46 (d, J
= 15.9 Hz, 1H), 5.45 (s, 1H), 5.24 (d, J = 9.0 Hz, 1H), 5.04 (d, J = 6.1
Hz, 1H), 3.87 (dd, J = 12.2, 1.9 Hz, 1H), 3.82 (dd, J = 9.1, 9.1 Hz,
1H), 3.73 (dd, J = 8.7, 4.4 Hz, 1H), 3.70 (dd, J = 12.0, 5.4 Hz, 1H),
3.60 (dd, J = 15.1, 5.7 Hz, 1H), 3.56–3.51 (m, 1H), 3.54 (dd, J = 8.9,
8.9 Hz, 1H), 3.48 (d, J = 9.2 Hz, 1H), 3.42 (ddd, J = 13.5, 11.3, 4.1
Hz, 1H), 3.28 (d, J = 14.2 Hz, 1H), 3.21 (dd, J = 14.6, 7.0 Hz, 1H),
3.05 (ddd, J = 13.4, 4.4, 4.4 Hz, 1H), 2.94 (d, J = 14.1 Hz, 1H), 2.89
(d, J = 16.5 Hz, 1H), 2.81 (ddd, J = 13.4, 6.4, 1.8 Hz, 1H), 2.55–2.38
(m, 3H), 2.24 (d, J = 16.5 Hz), 1.94 (ddd, J = 17.6, 7.9, 4.3 Hz, 1H),
1.86 (ddd, J = 17.5, 8.3, 4.3 Hz, 1H), 1.69–1.55 (m, 2H), 1.45–1.38
89.4, 81.1, 78.5, 73.8, 70.9, 62.5, 60.9, 57.4, 56.6, 52.8, 43.4, 42.7,
37.2, 34.8, 32.7, 32.4, 27.6, 26.6, 24.7, 24.5; HRMS (QE Orbitrap)
+
m/z: [M+H] calc’d for C50
H
58
N
9
O
11 960.4250; found 960.4254.
1
5
11b
8
(
1 S,1 R,1 S,12S,E)-12-((1H-imidazol-4-yl)methyl)-1 -bromo-
-hydroxy-1 -(3-(1-((2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-
6
1
7
(
hydroxymethyl)tetrahydro-2H-pyran-2-yl)-1H-1,2,3-triazol-4-
2
3
5
6
11 11b
1
yl)propyl)-1 ,1 ,1 ,1 ,1 ,1 -hexahydro-1 H-10,13-diaza-1(1,5)-
indolizino[8,7-b]indola-3,7(1,3)-dibenzenacyclotetradecaphan-4-
3
ene-1 ,11,14-trione (S11)
Synthesized according to Procedure F with 13 µmol of 43. Puri-
fied by preparative HPLC – see SI for conditions. White solid. TFA
1
salt: 1.5 mg [1.3 µmol, 10% yield]. H NMR (CD
3
OD, 500 MHz): δ
8
.63 (s, 1H), 7.56 (s, 1H), 7.53 (s, 1H), 7.50 (s, 1H), 7.41 (d, J = 7.5
Hz, 1H), 7.38 (dd, J = 7.5, 7.5 Hz, 1H), 7.22 (d, J = 8.5 Hz, 1H), 7.20
d, J = 8.5 Hz, 1H), 7.08 (d, J = 7.4 Hz, 1H), 6.97 (s, 1H), 6.92 (s,
H), 6.78 (d, J = 8.2 Hz, 1H), 6.62 (d, J = 8.1 Hz, 1H), 6.52 (br s),
5.50 (s, 1H), 5.36 (d, J = 9.1 Hz, 1H), 5.05 (d, J = 6.2, 1H), 4.28 (dd,
J = 9.0, 5.2 Hz, 1H), 3.88 (d, J = 11.9 Hz, 1H), 3.83 (dd, J = 9.2, 9.2
Hz, 1H), 3.72 (dd, J = 12.1, 5.5 Hz, 1H), 3.57–3.47 (m, 4H), 3.42–
1
3
(
1
3
(m, 2H), 1.32–1.25 (m, 3H); C NMR (CD OD, 126 MHz): δ 177.9,
176.7, 172.7, 171.7, 154.5, 140.2, 138.8, 138.4, 131.9, 131.1, 130.9,
130.8, 130.7, 130.1, 129.9, 129.3, 128.1, 127.8, 125.1, 123.0, 122.5,
120.4, 119.0, 115.3, 112.5, 108.7, 89.3, 81.0, 78.5, 73.8, 70.9, 62.4,
59.5, 55.8, 51.8, 49.8, 49.6, 47.5, 42.3, 41.0, 40.2, 36.7, 35.7, 34.4,
+
3
=
1
.83 (m, 1H), 3.22 (d, J = 15.7 Hz, 1H), 2.99–2.95 (m, 1H), 2.96 (d, J
14.7 Hz, 1H), 2.84–2.77 (m, 1H), 2.56–2.43 (m, 5H), 2.28 (d, J =
6.8 Hz, 1H), 1.70–1.64 (m, 1H), 1.61–1.54 (m, 1H), 1.38–1.26 (m,
30.9, 27.4, 26.7, 24.4, 23.6; HRMS (QE Orbitrap) m/z: [M+H] calc’d
for C49H N O11 918.4032; found 918.4036.
3-((1 S,1 R,11 S,13S,E)-1 ,12,15-trioxo-1 -(3-(1-
56 7
1 5 1b 3
1
1
3
6H); C NMR (CD
3
OD, 126 MHz): δ 175.9, 171.9, 171.4, 154.4,
((2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-
2
3
5
6
11 11b
1
1
1
6
48.4, 140.4, 139.2, 136.9, 135.0, 132.4, 131.7, 131.3, 131.2, 131.2,
31.1, 131.0, 130.1, 130.0, 129.4, 128.6, 127.9, 125.8, 125.8, 122.4,
21.4, 118.1, 115.5, 114.1, 113.4, 108.1, 89.4, 81.1, 78.5, 73.9, 70.9,
2H-pyran-2-yl)-1H-1,2,3-triazol-4-yl)propyl)-1 ,1 ,1 ,1 ,1 ,1
-
1
hexahydro-1 H-7-oxa-11,14-diaza-1(1,5)-indolizino[8,7-b]indola-
3(1,3),8(1,4)-dibenzenacyclopentadecaphan-4-en-13-yl)propanoic
acid (S14)
2.4, 60.5, 53.2, 51.8, 47.5, 42.1, 41.8, 41.0, 36.0, 35.6, 33.1, 28.1,
+
26.6, 24.7, 24.4; HRMS (QE Orbitrap) m/z: [M+H] calc’d for
Synthesized according to Procedure F beginning with 11 µmol of
C
50
H
55BrN
9
O
9
1004.3301; found 1004.3259.
40. Purified by preparative HPLC – see SI for conditions. White
1
5
10b
6
8
1
(
1 S,1 R,1 S,12S,E)-7 -hydroxy-12-((R)-1-hydroxyethyl)-1 -
solid. 3 mg [3.3 µmol, 30% yield]. H NMR (DMSO-d
6
, 500 MHz): δ
1
methoxy-1 -(3-(1-((2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-
hydroxymethyl)tetrahydro-2H-pyran-2-yl)-1H-1,2,3-triazol-4-
yl)propyl)-1 ,1 ,1 ,1 ,1 ,1 -hexahydro-10,13-diaza-1(1,5)-
7.74 (d, J = 8.5 Hz, 1H), 7.77 (s, 1H), 7.50 (d, J = 8.0 Hz, 1H), 7.41
(dd, J = 7.6, 7.6 Hz, 1H), 7.34 (d, J = 8.4 Hz, 1H), 7.27 (s, 1H), 7.26
(d, J = 6.8 Hz, 1H), 7.19–7.16 (m, 1H), 7.08 (dd, J = 7.5, 7.5, 0.8 Hz,
1H), 6.98 (d, J = 8.6 Hz, 1H), 6.69 (d, J = 8.4 Hz, 1H), 6.64 (d, J =
15.9, 1H), 5.98 (ddd, 15.8, 7.3, 5.9 Hz, 1H), 5.47 (s, 1H), 5.15 (d, J =
9.2 Hz, 1H), 5.12 (d, J = 5.7 Hz, 1H), 4.97 (dd, J = 12.4, 7.2 Hz, 1H),
4.46 (ddd, J = 7.8, 7.8, 7.8 Hz, 1H), 4.31 (dd, J = 12.3, 5.5 Hz, 1H),
4.86 (dd, J = 12.1 Hz, 1H), 3.84 (dd, J = 9.1, 9.1 Hz, 1H), 3.69 (dd, J
= 12.1, 5.4 Hz, 1H), 3.54 (dd, J = 8.9, 8.9 Hz, 1H), 3.51 (ddd, J = 9.6,
5.4, 2.2 Hz, 1H), 3.47 (d, J = 8.9 Hz, 1H), 3.44–3.40 (m, 1H), 3.03
(ddd, J = 16.2, 5.9, 2.2 Hz, 1H), 2.98 (d, J = 12.3 Hz, 1H), 2.64 (dd, J
= 7.1, 7.1 Hz, 2H), 2.57–2.52 (m, 1H), 2.55 (d, J = 16.8 Hz, 1H),
2.50–2.44 (m, 1H), 2.36 (d, J = 16.8 Hz, 1H), 1.91 (ddd, J = 16.5, 7.9,
3.3 Hz, 1H), 1.87–1.80 (m, 1H), 1.78–1.70 (m, 1H), 1.64–1.55 (m,
(
1
2
3
5
6
10b
pyrrolo[2,1-a]isoquinolina-3,7(1,3)-dibenzenacyclotetradecaphan-
3
4
-ene-1 ,11,14-trione (S12)
Synthesized according to Procedure F with 15 µmol 42. Purified
by preparative HPLC – see SI for conditions. White solid. 9 mg [10
1
µmol, 67% yield]. H NMR (CD
3
OD, 500 MHz): δ 7.63 (s, 1H), 7.50
(s, 1H), 7.34 (ddd, J = 8.1, 1.5, 1.5 Hz, 1H), 7.33 (d, J = 2.8 Hz, 1H),
7
1
6
.30 (d, J = 7.6 Hz, 1H), 7.29 (d, J = 8.7 Hz, 1H), 7.24 (d, J = 7.2 Hz,
H), 7.04 (ddd, J = 7.0, 1.5, 1.5 Hz, 1H), 7.01 (d, J = 2.2 Hz, 1H),
.84 (dd, J = 8.7, 2.7 Hz, 1H), 6.80, (dd, J = 8.1, 2.1 Hz, 1H), 6.74 (d,
J = 2.6 Hz, 1H), 6.63 (d, J = 8.1 Hz, 1H), 6.52 (d, J = 15.9 Hz, 1H),
6.46 (ddd, J = 15.7, 6.2, 6.2 Hz, 1H), 5.51 (d, J = 9.3 Hz, 1H), 5.36 (s,
ACS Paragon Plus Environment

Page 17 of 20
The Journal of Organic Chemistry
1
3
3
H), 1.45 (ddd, J = 13.5, 13.5, 3.8 Hz, 1H), 1.34–1.23 (m, 3H);
C
– see SI for conditions. TFA salt: 22 mg [23 µmol, 13% yield over
1
three steps]. H NMR (DMSO-d , 600 MHz): δ 8.88 (d, J = 7.7 Hz,
6
1
2
3
4
5
6
7
8
9
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
2
2
2
3
3
3
3
3
3
3
3
3
3
4
4
4
4
4
4
4
4
4
4
5
5
5
5
5
5
5
5
5
5
6
NMR (CD
3
OD, 126 MHz): δ 173.7, 172.2, 171.6, 169.7, 155.5, 137.4,
1
1
7
4
37.1, 137.0, 132.8, 130.2, 129.7, 129.4, 129.3, 128.9, 127.6, 126.4,
1H), 8.84 (br s, 1H), 8.77 (br s, 1H), 7.56 (dd, J = 4.4, 4.4 Hz, 1 H),
7.48 (d, J = 7.7 Hz, 1H), 7.36 (d, J = 8.5 Hz, 1H), 7.26–7.24 (m, 1H),
7.26 (dd, J = 7.2, 7.2 Hz, 2H), 7.20 (d, J = 7.1 Hz, 2H), 7.19 (dd, J =
7.1, 7.1 Hz, 1H), 7.13 (d, J = 8.2 Hz, 2H), 7.13 (d, J = 7.6 Hz, 1H),
6.87 (d, J = 8.3 Hz, 2H), 6.70 (d, J = 15.7 Hz, 1H), 6.56–6.51 (m,
1H), 48.3 (s, 1H), 4.62 (ddd, J = 6.7, 6.7, 6.7 Hz, 1H), 4.30–4.26 (m,
1H), 4.11 (dd, J = 8.9, 2.9 Hz, 1H), 3.90–3.89 (m, 1H), 3.79–3.77 (m,
1H), 3.79–3.76 (m, 1H), 3.75–3.73 (m, 1H), 3.55–3.49 (m, 2H), 3.37–
3.34 (m, 1H), 3.34–3.29 (m, 1H), 3.00–2.97 (m, 1H), 2.97–2.94 (m,
1H), 2.92 & 2.78 (AB quartet, J = 16.8 Hz, 2H), 2.85–2.81 (m, 1H),
2.77–2.74 (m, 1H), 2.72 (dd, J = 2.3, 2.3 Hz, 1H), 2.62–2.57 (m, 1H),
2.49 (s, 3H), 2.37 & 2.32 (AB quartet, J = 16.3 Hz, 2H), 2.13–2.08
(m, 1H), 2.11 (br s, 2H), 2.01–1.97 (m, 1H), 1.98–1.94 (m, 1H), 1.91–
1.87 (m, 1H), 1.78–1.76 (m, 2H), 1.74–1.70 (m, 1H), 1.57–1.52 (m,
1H), 1.54–1.51 (m, 1H), 1.49–1.44 (m, 2H), 1.39–1.30 (m, 2H), 1.33
26.2, 125.6, 121.7, 121.0, 119.2, 117.4, 114.8, 111.1, 106.7, 87.8,
9.6, 77.1, 72.3, 69.5, 63.9, 61.0, 57.8, 51.6, 50.8, 48.4, 44.2, 42.1,
+
0.8, 40.0, 36.0, 34.1, 28.7, 27.0, 25.3, 23.2, 22.8; MS m/z: [M+H]
calc’d for C49
H
56
N
7
O
11 918.4; found 918.5. HRMS (QE Orbitrap) m/z:
H N O
55 7
+
[
M+Na] calc’d for C49 11Na 940.3852; found 940.3859.
tert-Butyl (S)-(1-phenoxy-3-phenylpropan-2-yl)carbamate
Phenol [4.7 g, 50 mmol] in 48 mL of dry DMF was cannulated into
a suspension of NaH [2.0 g, 50 mmol] in 48 mL of dry DMF and
stirred for 5 min. tert-Butyl (S)-4-benzyl-1,2,3-oxathiazolidine-3-
carboxylate 2,2-dioxide [40 mmol], dissolved in 96 mL of dry DMF,
was cannulated into the reaction flask. The reaction was monitored
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
for completion by H-NMR. The reaction was quenched by addition
of 0.25 N HCl, and the mixture was then diluted with EtOAc. The
layers were separated, and the aqueous layer was extracted 2x with
EtOAc. The combined organic layers were washed 2x with 1N HCl,
1
3
6
(d, J = 6.6 Hz, 3H); C NMR (DMSO-d , 151 MHz): δ 172.7, 171.7,
3
x 1N NaOH, and 1x brine. The organic layer was dried with MgSO
4
171.2, 169.1, 168.6, 157.1, 144.5, 138.4, 137.0, 134.6, 131.7, 131.6,
129.6, 129.2, 128.8, 127.8, 126.6, 125.9, 123.5, 122.1, 114.9, 84.2,
71.3, 69.3, 68.4, 60.6, 55.8, 50.3, 49.1, 47.9, 46.6, 41.8, 41.2, 39.1,
and concentrated in vacuo. Chromatographed on silica using 10%
EtOAc/hexanes to provide tert-Butyl (S)-(1-phenoxy-3-phenylpropan-
2
-yl)carbamate as a white solid [6.2 g, 18.9 mmol]. 47% from com-
37.5, 36.9, 36.2, 35.2, 31.0, 30.7, 28.9, 26.9, 24.1, 23.8, 18.1, 15.6;
1
+
mercial L-phenylalaninol (4 steps). H NMR (CDCl
3
, 500 MHz): δ
HRMS (QE Orbitrap) m/z: [M+H] calc’d for C50
H
61
N
6
O
6
841.4647;
7
1
1
3
.30–7.26 (m, 5H), 7.21 (d, J = 7.2 Hz, 2H), 6.97 (dd, J = 7.3, 7.3 Hz,
found 841.4644.
3
a
8b
12
2
H), 6.89 (d, J = 8.4 Hz, 2H), 4.96 (d, J = 8.3 Hz, 1H), 4.19–4.12 (m,
H), 3.89 (dd, J = 9.4, 3.9 Hz, 1H), 3.86 (dd, J = 9.3, 3.5 Hz, 1H),
.02 (dd, J = 13.2, 6.3 Hz, 1H), 2.98 (dd, J = 13.0, 8.3 Hz, 1H), 1.43
(S)-N-((1 S,1 R,1 S,8S,13S,E)-8-benzyl-1 ,10,12,16-tetraoxo-
3a
1 2 3 3a 4 8b
1 -(pent-4-yn-1-yl)-1 ,1 ,1 ,1 ,1 ,1 -hexahydro-6-oxa-9,17-diaza-
1(6,1)-indeno[1,2-b]pyrrola-11(2,1)-pyrrolidina-5(1,4)-
benzenacyclononadecaphan-2-en-13-yl)-2-
1
3
(s, 9H); C NMR (CDCl
129.6, 128.7, 126.6, 121.2, 114.6, 79.7, 67.7, 51.4, 37.9, 28.6; HRMS
3
, 126 MHz): δ 158.7, 155.4, 137.9, 129.7,
(methylamino)propanamide (49b)
+
(
DART-Orbitrap) m/z: [M+H] calc’d for C20
28.1899.
S)-1-phenoxy-3-phenylpropan-2-amine
Pure tert-Butyl (S)-(1-phenoxy-3-phenylpropan-2-yl)carbamate
[6.2 g, 18.9 mmol] was dissolved in 160 mL DCM and cooled to 0 ºC.
0 mL of TFA was added to the reaction flask under argon. The reac-
H
26NO
3
328.1907; found
Isolated from previous reaction. TFA salt: 38 mg [40 µmol, 21%
1
3
yield over three steps]. H NMR (DMSO-d
6
, 600 MHz): δ 8.85 (br s,
(
1H), 8.75 (br s, 1H), 8.65 (d, J = 7.7 Hz, 1H), 7.90 (d, J = 6.8 Hz,
1H), 7.86 (dd, J = 4.4, 4.4 Hz, 1H), 7.64 (d, J = 7.9 Hz, 1H), 7.31–
7.28 (m, 1H), 7.27–7.25 (m, 1H), 7.25–7.24 (m, 1H), 7.22–7.21 (m,
1H), 7.21 (s, 1H), 7.11 (d, J = 8.3 Hz, 2H), 6.40–6.35 (m, 1H), 6.27
(d, J = 15.7 Hz, 1H), 4.55 (s, 1H), 4.34 (ddd, J = 9.2, 9.2, 2.1 Hz, 1H),
4.22 (dd, J = 8.2, 2.0 Hz, 1H), 4.09–4.08 (m, 1H), 4.09–4.08 (m, 1H),
3.97 (dd, J = 12.9, 7.1 Hz, 1H), 3.80–3.77 (m, 1H), 3.45 (dd, J = 14.8,
3.4 Hz, 1H), 3.38–3.35 (m, 1H), 3.34–3.32 (m, 1H), 3.29–3.26 (m,
1H), 3.29–3.26 (m, 1H), 3.14 (ddd, J = 8.2, 8.2, 8.2 Hz, 1H), 2.97–
2.93 (m, 1H), 2.95 & 2.82 (AB quartet, J = 16.4 Hz, 2H), 2.92–2.89
4
tion was monitored by TLC. After reaction completion, solvents were
removed and the residue partitioned between ethyl ether and 1N
NaOH; the layers were then separated. The ether layer was washed
2x with 1N NaOH and 1x with brine. The organic layer was dried
with MgSO
4
and concentrated in vacuo to give (S)-1-phenoxy-3-
phenylpropan-2-amine as a pale yellow, waxy solid [4.2 g, 18.5
!"
1
(
(
m, 1H), 2.80–2.77 (m, 1H), 2.72 (dd, J = 2.0, 2.0 Hz, 1H), 2.54–2.51
m, 1H), 2.53 (dd, J = 4.7, 4.7 Hz, 3H), 2.38 (s, 2H), 2.16–2.13 (m,
3 3
mmol]. 98% yield. [α] = +18.5º, c = 1.34, CHCl . H NMR (CDCl ,
!
5
7
6
00 MHz): δ 7.42–7.29 (m, 7H), 7.05 (ddd, J = 7.3, 1.0, 1.0 Hz, 1H),
.02–6.98 (m, 2H), 3.99 (dd, J = 9.0, 4.3 Hz, 1H), 3.87 (dd, J = 8.9,
.6 Hz, 1H), 3.51 (dddd, J = 7.7, 6.1, 6.1, 4.4 Hz, 1H), 3.00 (dd, J =
1H), 2.11 (ddd, J = 5.4, 5.4, 1.8 Hz, 2H), 2.07–2.04 (m, 1H), 1.91–
1.87 (m, 1H), 1.81–1.75 (m, 1H), 1.62–1.58 (m, 1H), 1.60–1.53 (m,
1
3
2H), 1.49–1.45 (m, 1H), 1.42–1.38 (m, 1H), 1.42–1.32 (m, 2H), 1.36–
13.4, 5.7 Hz, 1H), 2.78 (dd, J = 13.3, 8.0 Hz, 1H), 1.55 (br s, 2H);
C
13
1
6
.32 (m, 1H), 1.30 (d, J = 6.8 Hz, 3H); C NMR (DMSO-d , 151
NMR (CDCl
20.8, 114.5, 72.1, 52.0, 40.6; HRMS (DART-Orbitrap) m/z: [M+H]
calc’d for C15 18NO 228.1383; found 228.1379.
E)-3-(3-((3S,6S,16S)-16-(diethoxymethyl)-3-methyl-4,9,14-
3
, 126 MHz): δ 158.8, 138.5, 129.4, 129.2, 128.5, 126.4,
+
MHz): δ 172.0, 171.4, 170.7, 168.4, 156.6, 143.9, 138.2, 137.9, 137.6,
32.2, 132.2, 131.0, 129.4, 129.0, 128.8, 128.0, 126.1, 125.9, 123.8,
122.8, 84.0, 71.1, 70.5, 68.2, 58.4, 55.6, 49.6, 48.8, 47.3, 45.9, 42.4,
1
1
H
(
4
1
8
1.6, 38.9, 37.4, 36.6, 36.3, 36.2, 31.1, 30.7, 29.3, 26.9, 23.6, 23.6,
8.0, 15.4; HRMS (QE Orbitrap) m/z: [M+H] calc’d for C50H N O
61 6 6
41.4647; found 841.4644.
trioxo-16-(pent-4-yn-1-yl)-6-((S)-2-(((S)-1-phenoxy-3-
phenylpropan-2-yl)carbamoyl)pyrrolidine-1-carbonyl)-2,5,10,13-
tetraazaheptadecan-17-yl)phenyl)allyl isobutyl carbonate (48)
+
Dimer Product 50: Synthesized according to Procedure G using
Synthesized according to Procedure A with 0.30 mmol of (+)-5.
10 mg of 49a. Purified by preparative HPLC – see SI for conditions.
Carried forward without purification.
1
White solid. Bis-TFA salt: 4.5 mg [2.4 µmol, 40% yield]. H NMR
(
E)-3-(3-(((3S)-2-hydroxy-1-(2-((S)-4-((S)-2-
(
7
DMSO-d
.61 (dd, J = 4.7, 4.7 Hz, 1H), 7.49 (d, 7.8 Hz, 1H), 7.40 (d, J = 8.6
Hz, 1H), 7.28–7.23 (m, 3H), 7.23–7.17 (m, 3H), 7.14–7.12 (m, 4H),
.87 (d, J = 8.2 Hz, 2H), 6.72 (d, J = 15.5 Hz, 1H), 6.53 (ddd, J =
15.5, 8.5, 5.2 Hz, 1H), 4.84 (s, 1H), 4.62 (dd, J = 14.0, 6.8 Hz, 1H),
6
, 500 MHz): δ 8.90 (d, J = 7.7 Hz, 1H), 8.81 (br s, 2H),
(methylamino)propanamido)-5-oxo-5-((S)-2-(((S)-1-phenoxy-3-
phenylpropan-2-yl)carbamoyl)pyrrolidin-1-
yl)pentanamido)ethyl)-5-oxo-3-(pent-4-yn-1-yl)pyrrolidin-3-
yl)methyl)phenyl)allyl isobutyl carbonate (S35)
Synthesized according to Procedure B starting with 200 mg of
crude material. Diastereomeric mixture was carried forward without
purification.3
6
4
4
.31–4.24 (m, 1H), 4.10 (dd, J = 9.0, 3.2 Hz, 1H), 3.88 (dd, J = 9.5,
.6 Hz, 1H), 3.78–3.72 (m, 2H), 3.55–3.45 (m, 2H), 3.02–2.93 (m,
a
8b
12
2
1H), 2.95 (dd, J = 18.7, 4.6 Hz, 1H), 2.90–2.80 (m, 1H), 2.83 (dd, J =
13.8, 9.3 Hz, 1H), 2.79–2.76 (m, 2H), 2.37 (d, J = 16.2 Hz, 1H), 2.32
(d, J = 16.2 Hz, 1H), 2.23 (dd, J = 6.2, 6.2 Hz, 1H), 2.16–2.07 (m,
a(S)-N-((1 S,1 R,1 S,8S,13S,E)-8-benzyl-1 ,10,12,16-tetraoxo-
3
1
2
3
3a
4
8b
1
1
-(pent-4-yn-1-yl)-1 ,1 ,1 ,1 ,1 ,1 -hexahydro-6-oxa-9,17-diaza-
(8,1)-indeno[1,2-b]pyrrola-11(2,1)-pyrrolidina-5(1,4)-
1
1
H), 2.02–1.93 (m, 1H), 1.92–1.85 (m, 1H), 1.79–1.75 (m, 2H), 1.73–
benzenacyclononadecaphan-2-en-13-yl)-2-
(methylamino)propanamide (49a)
Synthesized with a modified General Procedure C with crude mate-
rial from above: 1:1 TFA/TFE, 5 mM. Purified by preparative HPLC
.67 (m, 1H), 1.53–1.46 (m, 3H), 1.43–1.28 (m, 4H), 1.33 (d, J = 6.87
13
6
Hz, 1H); C NMR (DMSO-d , 126 MHz): δ 172.6, 171.8, 171.2,
1
68.8, 168.3, 157.2, 144.8, 138.5, 137.2, 134.9, 131.8, 131.7, 129.6,
129.2, 128.2, 128.1, 126.9, 126.1, 124.0, 122.4, 115.0, 77.9, 69.6,
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8.6, 65.5, 60.7, 56.0, 50.3, 49.3, 48.4, 46.8, 42.2, 41.3, 40.4, 37.9,
1009.
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8)
Watkins, A. M.; Arora, P. S. Eur. J. Med. Chem. 2015, 94, 480–
488.
Neduva, V.; Linding, R.; Su-Angrand, I.; Stark, A.; De Masi, F.;
Gibson, T. J.; Lewis, J.; Serrano, L.; Russell, R. B. PLoS Biol.
2005, 3 (12), 1–10.
1
2
3
4
5
6
7
8
9
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
2
2
2
3
3
3
3
3
3
3
3
3
3
4
4
4
4
4
4
4
4
4
4
5
5
5
5
5
5
5
5
5
5
6
37.2, 36.4, 35.8, 31.2, 30.8, 29.1, 27.2, 24.2, 23.8, 18.8, 15.6; HRMS
+
(
ESI-TOF) m/z: [M+Na] calc’d for C100
H
118
N
12
O
12Na 1702.8918;
(
found 1702.8978.
Dimer Product 6: Synthesized according to Procedure G using 10
mg of 49b. Purified by preparative HPLC – see SI for conditions.
1
(9)
Van Roey, K.; Uyar, B.; Weatheritt, R. J.; Dinkel, H.; Seiler,
M.; Budd, A.; Gibson, T. J.; Davey, N. E. Chem. Rev. 2014, 114
White solid. Bis-TFA salt: 6.8 mg [3.6 µmol, 60% yield]. H NMR
(
7
7
DMSO-d
6
, 500 MHz): δ 8.81 (br s, 1H), 8.76 (br s, 1H), 8.65 (d, J =
.7 Hz, 1H), 7.91 (d, J = 6.6 Hz, 1H), 7.88 (dd, J = 4.1, 4.1 Hz, 1H),
.65 (d, J = 7.9 Hz, 1H), 7.37–7.34 (m, 1H), 7.36 (s, 1H), 7.29–7.22
(
13), 6733–6778.
Nevola, L.; Giralt, E. Chem. Commun. 2015, 51 (16), 3302–
315.
Bock, J. E.; Gavenonis, J.; Kritzer, J. A. ACS Chem. Biol. 2013,
(3), 488–499.
(10)
3
(
m, 5H), 7.20 (s, 1H), 7.11 (d, J = 8.2 Hz, 2H), 6.87 (d, J = 8.3 Hz,
2H), 6.40–6.35 (m, 1H), 6.27 (d, J = 15.7 Hz, 1H), 4.54 (s, 1H), 4.33
dd, J = 8.5, 8.5 Hz, 1H), 4.22 (dd, J = 8.1, 2.3 Hz, 1H), 4.09 (d, J =
.9 Hz, 1H), 3.96 (dd, J = 12.7 Hz, 1H), 3.79–3.75 (m, 1H), 3.64–3.60
m, 1H), 3.15–3.11 (m, 1H), 2.96–2.89 (m, 2H), 2.83–2.76 (m, 2H),
.37 (br s, 2H), 2.24 (dd, J = 6.5, 6.5 Hz, 2H), 2.19–2.12 (m, 1H),
(
11)
8
(
7
(
2
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
(12)
13)
Yudin, A. K. Chem. Sci. 2015, 6 (1), 30–49.
Miller, S. J.; Blackwell, H. E.; Grubbs, R. H. J. Am. Chem. Soc.
1996, 118 (40), 9606–9614.
Blackwell, H. E.; Grubbs, R. H. Angew. Chem. Int. Ed. 1998, 37
(23), 3281–3284.
Isidro-Llobet, A.; Murillo, T.; Bello, P.; Cilibrizzi, A.;
Hodgkinson, J. T.; Galloway, W. R.; Bender, A.; Welch, M.;
Spring, D. R. Proc. Natl. Acad. Sci. U.S.A. 2011, 108 (17),
(
(14)
(15)
2.08–2.03 (m, 1H), 1.91–1.86 (m, 1H), 1.80–1.74 (m, 1H), 1.65–1.47
(m, 5 H) 1.41–1.30 (m, 4H), 1.30 (d, J = 7.0 Hz, 3H); HRMS (ESI-
+
TOF) m/z: [M+Na] calc’d for C100
H
118
N
12
O
12Na 1702.8918; found
1
702.8912.
6
793–6798.
(16)
Beckmann, H. S. G.; Nie, F.; Hagerman, C. E.; Johansson, H.;
Tan, Y. S.; Wilcke, D.; Spring, D. R. Nat. Chem. 2013, 5
ASSOCIATED CONTENT
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October), 861–867.
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P. G. J. Am. Chem. Soc. 2002, 113 (24), 9391–9392.
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1105–1111.
The Supporting Information is available free of charge on the
ACS Publications website: Fluorescence polarization assay, Ca-
co-2 permeability screenings, computational procedures, & spec-
troscopic data (PDF).
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(
(
(
19)
20)
21)
AUTHOR INFORMATION
Corresponding Author
*
harran@chem.ucla.edu
Present Addresses
Koch Institute for Integrative Cancer Research at MIT, 500
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23)
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25)
(26)
Hopkins, B. A.; Smith, G. F.; Sciammetta, N. Org. Lett. 2016,
1
8 (16), 4072–4075.
(
Le, D. N.; Riedel, J.; Kozlyuk, N.; Martin, R. W.; Dong, V. M.
Org. Lett. 2017, 19 (1), 114–117.
Gartner, Z. J.; Tse, B. N.; Grubina, R.; Doyon, J. B.; Snyder, T.
M.; Liu, D. R. Science 2004, 305 (5690), 1601–1605.
Li, X.; Gartner, Z. J.; Tse, B. N.; Liu, D. R. J. Am. Chem. Soc.
§
Main Street, Cambridge, MA 02139
∫ Center for Catalytic Hydrocarbon Functionalization, Institute for
Basic Science (IBS), Daejeon, 34141, Republic of Korea.
(
2
004, 126 (16), 5090–5092.
◊
Modeling and Informatics, Merck & Co., Inc., P. O. Box 2000,
Tse, B. N.; Snyder, T. M.; Shen, Y.; Liu, D. R. J. Am. Chem.
Soc. 2008, 130 (46), 15611–15626.
Rahway, NJ 07065
(
(
(
27)
28)
29)
White, T. R.; Renzelman, C. M.; Rand, A. C.; Rezai, T.;
McEwen, C. M.; Gelev, V. M.; Turner, R. A.; Linington, R. G.;
Leung, S. S. F.; Kalgutkar, A. S.; Bauman, J. N.; Zhang, Y.;
Liras, S.; Price, D. A.; Mathiowetz, A. M.; Jacobson, M. P.;
Lokey, R. S. Nat. Chem. Biol. 2011, 7 (11), 810–817.
Rand, A. C.; Leung, S. S. F.; Eng, H.; Rotter, C. J.; Sharma, R.;
Kalgutkar, A. S.; Zhang, Y.; Varma, M. V.; Farley, K. A.;
Khunte, B.; Limberakis, C.; Price, D. A.; Liras, S.; Mathiowetz,
A. M.; Jacobson, M. P.; Lokey, R. S. Medchemcomm 2012, 3
(10), 1282.
Notes
The authors declare no competing financial interest.
ACKNOWLEDGMENT
Funding was provided by the NIH (CA184772 to P.G.H.), a grant
from the University of California Cancer Research Coordinating
Committee, the D. J. & J. M. Cram Endowment, the Human Fron-
tier Science Program (LT001006/2015-C to F.M.), NSF Equip-
ment Grant CHE-1048804, and NIH instrumentation grant
Bockus, A. T.; Lexa, K. W.; Pye, C. R.; Kalgutkar, A. S.;
Gardner, J. W.; Hund, K. C. R.; Hewitt, W. M.; Schwochert, J.
A.; Glassey, E.; Price, D. A.; Mathiowetz, A. M.; Liras, S.;
Jacobson, M. P.; Lokey, R. S. J. Med. Chem. 2015, 58 (11),
1
S10OD016387-01. We thank Dr. Tristin Rose for helpful discus-
sions and guidance. Dr. Noelle Williams (UT Southwestern Med-
ical Center) is thanked for Caco2 permeability screens and expert
advice. We also thank Professor Neil Garg and Lucas Morrill
4
581–4589.
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Maianti, J. P.; McFedries, A.; Foda, Z. H.; Kleiner, R. E.; Du,
X. Q.; Leissring, M. A.; Tang, W.-J.; Charron, M. J.; Seeliger,
M. A.; Saghatelian, A.; Liu, D. R. Nature 2014, 511 (7507), 94–
(
UCLA) for use of their supercritical fluid chromatography in-
strument.
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8.
Wei, Q.; Harran, S.; Harran, P. G. Tetrahedron 2003, 59 (45),
8947–8954.
Zhao, H.; Negash, L.; Wei, Q.; LaCour, T. G.; Estill, S. J.;
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