Discovery of novel p-arylthio cinnamides as antagonists of leukocyte function-associated antigen-1/intracellular adhesion molecule-1 interaction. 1. Identification of an additional binding pocket based on an anilino diaryl sulfide lead

J . Med. Chem. 2000, 43, 4025-4040

4025

Discover y of Novel p-Ar ylth io Cin n a m id es a s An ta gon ists of Leu k ocyte

F u n ction -Associa ted An tigen -1/In tr a cellu la r Ad h esion Molecu le-1 In ter a ction . 1.

Id en tifica tion of a n Ad d ition a l Bin d in g P ock et Ba sed on a n An ilin o Dia r yl

Su lfid e Lea d

Gang Liu,*^,†J . T. Link,^†Zhonghua Pei,^†Edward B. Reilly,^†Sandra Leitza,^†Bach Nguyen,^§Kennan C. Marsh,^§

Gregory F. Okasinski,^†Thomas W. von Geldern,^†Mark Ormes,^‡Kerry Fowler,^‡and Mike Gallatin^‡

Metabolic Disease Research and Drug Analysis Department, Pharmaceutical Products Division, Abbott Laboratories,

Abbott Park, Illinois 60064-6098, and ICOS Corporation, 22021 20th Avenue SE, Bothel, Washington 98021

Received J une 28, 2000

The interaction between leukocyte function-associated antigen-1 (LFA-1), a member of the â₂-

integrin family of adhesion molecules, and intracellular adhesion molecule ICAM-1 (cd54) is

thought to play a critical role in the inflammatory process. On the basis of an anilino diaryl

sulfide screening lead 1, in combination with pharmacophore analysis of other screening hits,

we have identified an adjacent binding pocket. Subsequently, a p-ethenylcarbonyl linker was

discovered to be optimal for accessing this binding site. Solution-phase parallel synthesis enabled

rapid optimization of the cinnamides for this pocket. In conjunction with fine-tuning of the

diaryl substituents, we discovered a novel series of potent, nonpeptide inhibitors of LFA-1/

ICAM-1 interaction, exemplified by A-286982 (28h ), which has IC₅₀values of 44 and 35 nM in

an LFA-1/ICAM-1 binding assay and LFA-1-mediated cellular adhesion assay, respectively.

In tr od u ction

To date, most of the agents designed for blocking the

binding of LFA-1 to ICAM-1 have been monoclonal

antibody based.^6,7Several small molecules have been

reported to prevent cell adhesion via inhibition of

integrin activation, but they do not directly block

association of the purified proteins.^8,9More recently,

three accounts of structurally diverse small molecules

have been reported to prevent the binding of LFA-1 to

ICAM-1.^10-12Herein, we report the discovery of a series

of p-arylthio cinnamides that inhibit LFA-1 from bind-

ing to ICAM-1, interrupting the endothelial cell-leu-

kocyte adhesion process in vitro. These compounds are

potentially useful for the prophylaxis or treatment of

diseases in which leukocyte trafficking plays a role,

notably acute and chronic inflammatory diseases, auto-

immune diseases, tumor metastasis, allograft rejection,

and reperfusion injury.

Inflammation results from a cascade of events that

includes vasodilation accompanied by increased vascu-

lar permeability and exudation of fluid and plasma

proteins. Inflammatory mediators (chemokines such as

IL-8, MCP-1, MIP-1, RANTES, etc.) have chemotactic

activity for leukocytes and attract the circulating leu-

kocytes to localize and cross the vascular endothelium

at a precise location. This leukocyte recruitment is

accomplished by a process called cell adhesion.¹

Cell adhesion occurs through a coordinately regulated

series of steps that allow the leukocytes to first adhere

to a specific region of the vascular endothelium and then

cross the endothelial barrier to migrate to the inflamed

tissue.²These steps are mediated by families of adhesion

molecules such as integrins, Ig supergene family mem-

bers, and selectins which are expressed on the surface

of the circulating leukocytes and on the vascular en-

dothelial cells. Leukocyte function-associated antigen-1

(LFA-1, also termed CD11a/CD18) belongs to a member

of the â₂-integrin family of adhesion molecules, R_Lâ₂.

LFA-1 is thought to play a critical role in the inflam-

matory process by interacting with the intercellular

adhesion molecule ICAM-1 (CD54), an immunoglobin

expressed on endothelial cells, this interaction promotes

the migration of leukocytes rapidly into surrounding

tissue.³Thus, an agent which blocks the LFA-1/ICAM-1

interaction may suppress these early steps in the

inflammatory response.⁴Consistent with this back-

ground, ICAM-1 knockout mice have numerous abnor-

malities in their inflammatory responses.⁵

Our process of discovery began with the identification

of anilino diaryl sulfide 1 from screening a compound

library. It has low-micromolar potency against the

binding of LFA-1 to ICAM-1 (Scheme 1). Preliminary

SAR studies indicated that both the sulfur atom and

the anilino group are critical for the affinity of the

compound. Alkylation and acylation of the anilino group

generally attenuated the potency of the resulting com-

pounds (data not shown). Replacement of the sulfide

linkage with oxygen, carbonyl, and methylene groups

also reduced the affinities of the compounds (data not

shown). On the other hand, compound 2 was identified

from the same screening and found to moderately

antagonize the LFA-1/ICAM-1 interaction. Even though

compounds 1 and 2 likely have very different bound

conformations, it is intriguing to note the tolerance of

an N,N-dimethylaminoethyl group on the phenothiazine

template of 2, which might indicate the existence of an

additional binding pocket adjacent to the diaryl sulfide

* Address for correspondence: D-47R, AP-10, Abbott Laboratories,

100 Abbott Park Rd., Abbott Park, IL 60064-6098. Tel: 847-935-1224.

Fax: 847-938-1674. E-mail: Gang.Liu@abbott.com.

^†Metabolic Disease Research, Abbott Laboratories.

^§Drug Analysis Department, Abbott Laboratories.

^‡ICOS Corp.

Published on Web 10/19/2000

4026 J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 21

Liu et al.

Sch em e 1. Pharmacophore Analysis of Screening Leads of LFA-1/ICAM-1 Inhibitors

Sch em e 2^a

a

Reagents and conditions: (a) K₂CO₃, DMF, 70 °C, 83%; (b) SnCl₂, EtOH, reflux; (c) acryloyl chloride, Et₃N, CH₂Cl₂, 65% two steps;

(d) HNR₁R₂, MeOH, reflux, 55-90%; (e) LAH, THF, rt, 86%.

binding site. We decided to explore the tolerance of such

ortho substituents on a more flexible template, such as

diaryl sulfide 1.

From bromide 18, bis-cinnamide 19 was synthesized

through palladium-catalyzed Heck reaction with acry-

loylmorpholine (Scheme 4). Buchwald coupling¹⁵of the

same bromide with aminopropylmorpholine yielded the

A-ring functionalized anilino compound 22. Activation

of the alcohol 20 as a bromide and displacement of

which with secondary amines provided analogues 21

(Scheme 5). Similar Heck reaction of bromide 23 with

acryloylmorpholine gave the mono-cinnamide 24 (Scheme

6). Subsequent homologation and amidation yielded the

other series of bis-cinnamide 25.

Ch em istr y

Scheme 2 describes the synthesis of a typical diaryl

sulfide aniline compound 8 and the corresponding amide

precursor 7. Nucleophilic substitution of the 2-chloro-

nitrobenzene (4) with 2,4-dichlorobenzenethiol (3) yielded

the diaryl sulfide 5. The nitro group of 5 was then

reduced to the amine, which was subsequently acylated

with acryloyl chloride. Michael addition of the resulting

acrylamide 6 with secondary amines provided amides

7. Lithium aluminum hydride reduction of the amide 7

offered N,N-dialkyl(aminopropyl)amino diaryl sulfide 8.

In a similar fashion, diaryl sulfide aldehydes (both

ortho- and para-substituted) 10 were synthesized from

substituted chlorobenzaldehydes 9 and substituted ben-

zenethiols, such as 3 (Scheme 3). The meta-substituted

aldehyde 10 can be prepared through an Ullman

coupling¹³of benzenethiol 3 with 3-iodobenzoic acid (11),

followed by borane reduction of the resulting acid 12

and oxidation of thus formed alcohol. The aldehydes 10

were converted to the corresponding cinnamic acids 14

using the Doebner modification of the Knoevenagel

condensation with malonic acid.¹⁴Activation of acids as

acid chlorides, followed by condensation with primary

or secondary amines, provided the desired cinnamides

15-17. The o-aldehyde 10 was also subjected to a

standard reductive amination protocol to give amine 13.

Alternatively, the order of assembly may be reversed

to offer a greater flexibility for exploring the SAR of the

A-ring (Scheme 7). 4-Chloro-3-nitrocinnamic acid (26)

was coupled with a primary or secondary amine as

described before to give the corresponding amide 27. The

chlorine of 27 can then be displaced with virtually any

thiophenol to provide a library of analogues 28. Func-

tional groups on the A- and B-rings of the diaryl sulfide

28 can be further modified to produce new analogues,

such as amine 29. Sandmeyer reaction converted the

amine 29 to bromide 30, which can undergo Buchwald

coupling to give the anilino analogue 31.

B-Ring-modified cinnamides, such as 34 and 35, were

prepared from diaryl sulfide 32 through the same

palladium-mediated Heck coupling described for 18

(Scheme 8). The resultant aldehyde 33 can be oxidized

to the carboxylic acid 34 with silver oxide,¹⁶and

amidation of the resulting acid provided amide 35.

Novel p-Arylthio Cinnamides

J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 21 4027

Sch em e 3^a

a

Reagents and conditions: (a) K₂CO₃, DMF, 70 °C, ca. 85%; (b) KOH, H₂O, Cu, 2,4-dichlorothiophenol, reflux, 97%; (c) BH₃‚THF, rt,

97%; (d) Swern oxidation, 47%; (e) N,N-dimethylethylenediamine, NaBH₄, MeOH, 95%; (f) malonic acid, cat. piperidine, pyr, 110 °C, 91%;

(g) (COCl)₂, CH₂Cl₂, cat. DMF, rt; (h) HNR₁R₂, i-PrNEt₂, cat. DMAP, CH₂Cl₂, 90%; (i) 1 N NaOH, MeOH, 95%.

Sch em e 4^a

a

Reagents and conditions: (a) acryloylmorpholine, Pd(OAc)₂, P(o-tolyl)₃, Et₃N, DMF, 110 °C, 39%; (b) aminopropylmorpholine, Pd₂(dba)₃,

BINAP, NaOt-Bu, 18-C-6, toluene, 80 °C, 44%.

Sch em e 5^a

of ortho substituents on the diaryl sulfide template 1.

A synthetic strategy of using Michael addition of

secondary amines to the acrylamide 6 was devised to

allow access to both the amido and anilino analogues

sequentially. The acyclic dialkylaminoethyl amide ana-

logues were much weaker inhibitors than 1, with N,N-

dimethylamine (7a ) and N,N-di-n-propylamine (7b)

analogues exhibiting IC₅₀values close to 100 µM, and

N-methylbenzylamine (7c) analogue having an IC₅₀

value greater than 100 µM (Table 1). For the cyclic

amine analogues, five-membered pyrrolidine-derived

compound 7d did not showed any affinity, and six-

membered morpholine (7e) and N-methylpiperazine

(7f)-based analogues are well-tolerated, exhibiting only

a 5-10-fold decrease of potency compared with the

parent compound 1. When the amide of 7e was reduced

to aniline 8, more than an 85-fold boost of potency was

observed to give a submicromolar inhibitor of LFA-1/

ICAM-1 association. Notably, compound 13, derived

from changing the anilinopropyl linker of 8 to a methyl-

aminoethyl and the morpholine to the less optimal

dimethylamine, has a comparable IC₅₀value to that of

compound 1.

a

Reagents and conditions: (a) PBr₃, LiBr, DMF; (b) HNR₁R₂,

i-PrNEt₂, Ch₃CN, 87%.

Str u ctu r e-Activity Rela tion sh ip Discu ssion

The primary biochemical assay measures the ability

of the compound to block the interaction between the

integrin LFA-1 and its adhesion partner ICAM-1 in a

time-resolved fluorescent LFA-1/ICAM-1 biochemical

interaction assay. Biologically relevant activity of the

compounds can be confirmed using a cell-based adhesion

assay, which measures the ability of a compound to

block the adherence of J Y-8 cells (a human EBV-

transformed B cell line expressing LFA-1 on its surface)

to immobilized ICAM-1.

On the basis of the hypothesis that an additional

binding pocket adjacent to the compound 1 binding site

might be available, we decided to explore the tolerance

Once the existence of an additional binding space

4028 J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 21

Liu et al.

Sch em e 6^a

a

Reagents and conditions: (a) acryloylmorpholine, Pd(OAc)₂, P(o-tolyl)₃, Et₃N, DMF, 110 °C, 77%; (b) malonic acid, cat. piperidine,

pyr, 110 °C, 91%; (c) (COCl)₂, CH₂Cl₂, cat. DMF, rt; (d) HNRR₁, i-PrNEt₂, cat. DMAP, CH₂Cl₂, 90%.

Sch em e 7^a

a

Reagents and conditions: (a) HNRR₁, EDAC, DMF, 92%; (b) K₂CO₃, DMF, rt, 89%; (c) SnCl₂, EtOH, reflux; (d) tert-butyl nitrite,

CuBr₂, CH₃CN, 29%; (e) aminopropylmorpholine, Pd₂(dba)₃, BINAP, NaOt-Bu, 18-C-6, toluene, 80 °C, 44%.

Sch em e 8^a

a

Reagents and conditions: (a) acryloylmorpholine, Pd(OAc)₂, P(o-tolyl)₃, Et₃N, DMF, 110 °C, 77%; (b) Ag₂O, H₂O, 3 N NaOH, 71%; (c)

EDAC, HOBT, NH₄Cl, Et₃N, DMF, 74%.

adjacent to where compound 1 binds was established

by the improved potency exhibited by compound 8, we

started to search for different linkers and terminal

amines to fill that portion of the binding pocket. Because

of the availability of the intermediate aldehyde 10, and

the ease of synthesis and versatility for further ma-

nipulation, the ethenylcarbonyl group was examined as

a three-atom linker between the aromatic ring and the

terminal amino group. Interestingly, the o-cinnamic acid

itself 14a was almost equipotent as compound 1,

representing the first non-aniline-containing, low-mi-

cromolar inhibitor of the series (Table 2). The results

from the first small set of amides prepared from acid

14a were rather encouraging and offered direction for

optimizing this series of LFA-1/ICAM-1 inhibitors.

Hydrophobic amines, such as phenethylamine (15a ),

2-aminobenzothiazole (15b), and phenoxybenzylbenzy-

lamine (15c), yielded completely inactive analogues. In

contrast, amidation of the cinnamic acid 14a with

1-amino-6-hexanol yielded 15d , an inhibitor slightly

more potent than the acid 14a . Prompted by this critical

observation, other hydrophilic amino alcohols, amino

acids, and aminoalkylamines were examined with this

o-cinnamide template. Ethanolamine-derived cinnamide

15e is slightly less potent than the hexanolamine

analogue 15d. Similarly, 2,3-dihydroxypropylamine (15f)

and diethanolamine (15g) yielded analogues with com-

parable affinities to that of 15e. Compared with alcohol

15e, glycine-derived analogue 15h showed essentially

identical potency. When an amino group such as imi-

dazole (15i) was appended to the terminus of the

cinnamide, the potency of the resulting compound

remained the same as 15d . Neutral heterocyclic alky-

lamines, such as aminotetrahydrofurfuryl and amino-

propylpyrrolidine, produced cinnamides (15j, 15k) which

are also potent LFA-1/ICAM-1 antagonists. Similar to

15g, morpholine-derived tertiary cinnamide 15l exhib-

ited identical potency to 15d .

Encouraged by the early success of the o-cinnamides,

we surveyed the position preference of the cinnamide

relative to the arylthio group using the above-mentioned

preferred amines. In comparison to o-cinnamides, the

corresponding m-cinnamides are generally less active

against LFA-1/ICAM-1 interaction. The decrease of the

potency ranges from 15-fold (morpholino analogue 16d )

to 3-fold (aminopropylpyrrolidinone analogue 16e) and

somewhere in between for amino alcohols 16a and 16b

and aminoalkylamine derivative 16c (Table 3). For the

p-cinnamides, the reversed trend was observed. p-

Cinnamic acid 14b is equipotent to the ortho analogue

14a . For the amino alcohols, such as ethanolamine

(17a ), 1-amino-6-hexanol (17b), and diethanolamine

(17c), a trend of improvement over the corresponding

o-cinnamides was observed. Imidazole analogue 17d

exhibited comparable affinity to 15i, while aminopro-

pylpyrrolidinone 17e and morpholine 17f showed ap-

proximately 3-fold improvement as well. Using the

p-cinnamide template, the apparent preference of this

Novel p-Arylthio Cinnamides

J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 21 4029

Ta ble 1. Structure-Activity Relationships of

o-N,N-Dialkylpropyl(oxo)amino Diaryl Sulfides and

N,N-Dialkylethylaminomethyl Diaryl Sulfides

Ta ble 2. Structure-Activity Relationships of o-Cinnamido

Diaryl Sulfides

a

IC₅₀calculated using a mean of at least two measurements

(all duplicates) for 6 concentrations from 3.2 × 10^-8to 10^-4unless

otherwise noted.

series of inhibitors for six-membered cyclic amine was

briefly explored. The basic N-methylpiperazine (17g)

and N-pyridylpiperazine (17i) analogues were slightly

less potent than the morpholine analogue 17f. N-

Acetylpiperazine compound 17h turned out to be 3-fold

more potent than 17i, the most active cinnamide

examined so far.

During this process of searching for an agent suitable

for oral therapy of diseases related to inflammatory

response, we had successfully eliminated the metaboli-

cally liable aniline group of 1. The other potentially

troublesome features of this series of compounds are the

sulfide and cinnamide double bond. The pharmacoki-

netic behavior of the representative compound 17e of

this novel series of LFA-1/ICAM-1 inhibitors was ex-

amined, and the results were encouraging. In rats,

compound 17e was reasonably absorbed following 5 mg/

kg oral administration and achieved good plasma drug

level (t_1/2) 1.8 h, C_max) 1.08 µg/mL, AUC ) 3.26 µg‚

h/mL). The compound was not extensively metabolized

based on the rat microsome metabolism studies (data

not shown), suggesting the viability of both the sulfide

and the cinnamide double bond as parts of the drug

molecule. The calculated oral bioavailability (F) is 33%,

which indicates that this series of compounds has the

potential to be orally deliverable agents.

a

IC₅₀calculated using a mean of at least two measurements

(all duplicates) for 6 concentrations from 3.2 × 10^-8to 10^-4unless

otherwise noted.

marginal potency increase realized by switching from

o-cinnamides to p-cinnamides could be attributed to the

better fitting of the compounds in the receptor or the

presence of an additional chlorine on the p-cinnamides.

To address this question, some hybrid molecules car-

rying both o- and p-cinnamides were synthesized, and

results from this study are summarized in Table 4.

When the p- and o-cinnamides were situated on the two

opposing phenyl groups of the diaryl sulfide, exemplified

with compound 19, the resulting antagonist exhibited

only a slight increase of affinity over p-cinnamide 17f.

At the onset of the project, we had also examined

o-benzylamine 13 for filling this additional binding site.

Naturally, we explored the combination of o-benzyl-

At this stage, it was still unclear whether the p-

cinnamides bind to the same site as the o-cinnamides

or if they interact with yet another binding pocket. The

4030 J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 21

Liu et al.

Ta ble 3. Structure-Activity Relationships of m- and

p-Cinnamido Diaryl Sulfides

The results from positioning both o- and p-cinnamides

on the same aromatic ring turned out to be quite

different. Fixing the o-cinnamide as morpholinamide,

aminopropylpyrrolidinone as the p-cinnamide partner

rendered virtually inactive compound 25a , so did N-

acetylpiperazine (25b) and morpholine (25c) analogues.

The lack of synergy between o- and p-cinnamides

appears to indicate the existence of only one additional

binding pocket. We focused on the p-cinnamide as the

lead structure for further optimization based on its

better potency.

The three portions of the p-arylthio cinnamide were

examined separately to identify optimal substituents for

final assembly of a more potent inhibitor. Replacing the

chlorine on the central phenyl ring (B-ring) of 17e with

a nitro group led to a small increase of the potency of

the resulting compound 28a (Table 5). On the basis of

the existing SAR of the B- and C-rings (cinnamide

portion), the A-ring (p-arylthio) substituents were then

explored using compound 28b (IC₅₀) 0.090 µM), an

N-acetylpiperazine analogue, as the benchmark. The

main focus was to examine hydrophobic benzenethiols

as A-ring partners, along with some hydrophilic ben-

zenethiols as wild cards. The results from this exercise

seem to suggest that the binding pocket for the A-ring

consists primarily of hydrophobic amino acid residues.

2,3-Dichlorobenzenethiol yielded inhibitor 28c equi-

potent to the reference analogue 28b, so did 2-chloro

(28d ) and 2-methyl (28e) benzenethiols. To fully take

advantage of the hydrophobic interaction at the 2-posi-

tion of the A-ring, different alkyl substituents were

examined. An ethyl group (28g) was slightly better than

the methyl group, and the isopropyl group in turn

yielded a slightly more active analogue 28h (A-286982).

But the tert-butyl group offered compound 28i with

more than a 10-fold decrease in potency compared with

28h . In contrast to the incremental improvement seen

from 28c to 28h , the hydrophilic groups faired worse

than the hydrophobic groups. 2-Aminobenzenethiol

analogue 28f is 6-fold less active than the 2-chloro

compound 28e. N,N-Dimethylaminobenzenethiol de-

rivative 28k is 7-fold less active than its analogous 28h .

2-Formylbenzenethiol resulted in a very potent inhibitor

28j, although 2-carboxamidobenzenethiol 28l led to

almost complete negation of all the affinity we have

gained so far.

For the SAR of the B-ring, electron-donating groups,

such as amino (29) and morpholinopropylamino (31),

yielded much less active compounds. Other electron-

withdrawing groups, such as bromine (30) and formyl

(33), yielded analogues with similar potency as their

chlorine counterpart. The negatively charged carboxylic

acid (34) and neutral primary carboxamide group (35)

replacing the chlorine of 17e led to a complete loss of

potency against LFA-1/ICAM-1 interaction.

a

IC₅₀calculated using a mean of at least two measurements

(all duplicates) for 6 concentrations from 3.2 × 10^-8to 10^-4unless

otherwise noted.

amines with p-cinnamides on the two opposing phenyl

rings of the sulfide. Among the secondary amines

explored, morpholine (21a ), methylenedioxybenzylpip-

erazine (21c), and N-methylsarcocine (21d ) derivatives

are as active as the bis-cinnamide 19, while N-formylpip-

erazine analogue 21b is only marginally less active than

19. Interestingly, combining the o-anilino substituent

of 8 with p-cinnamide yielded the equally potent an-

tagonist 22 against LFA-1/ICAM-1 interaction as 19.

To confirm the cellular relevancy of inhibiting LFA-

1/ICAM-1 inaction in the primary assay, we devised a

secondary assay using an IL-8-stimulated J Y cell sys-

tem. A selective panel of compounds were examined, and

the results are shown in Table 6. In general, the IC₅₀

values derived from the primary assay agree well with

the data from the adhesion assay. For example, the most

active compound in the group, 28h , exhibited an IC₅₀

Novel p-Arylthio Cinnamides

J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 21 4031

Ta ble 4. Structure-Activity Relationships of Chimeras of Ortho- and Para-Disubstituted Cinnamido Diaryl Sulfides

a

IC₅₀calculated using a mean of at least two measurements (all duplicates) for 6 concentrations from 6.4 × 10^-9to 2 × 10^-5unless

otherwise noted.

value of 0.035 µM in the J Y-8 cell adhesion assay versus

0.044 µM in the LFA-1/ICAM-1 biochemical assay.

proximity to the one occupied by the original lead

compound and by identifying the preferred linker to

access that binding site. Rapid parallel optimization of

the different portions of the lead structure and combi-

nation of the preferred structural features led to the

identification of compound 28h , a potent representative

of this novel series of LFA-1/ICAM-1 interaction inhibi-

tors. Studies of the mechanism of the inhibition for this

Su m m a r y

A novel series of p-arylthio cinnamide-based inhibi-

tors of LFA-1/ICAM-1 interaction has been identified,

starting from an anilino diaryl sulfide screening lead,

by mapping out additional binding space in close

4032 J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 21

Liu et al.

Ta ble 5. Structure-Activity Relationships of A- and B-Ring Modification

Novel p-Arylthio Cinnamides

J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 21 4033

Ta ble 5. (Continued)

a

IC₅₀calculated using a mean of at least two measurements (all duplicates) for 6 concentrations from 6.4 × 10^-9to 2 × 10^-5unless

otherwise noted.

under nitrogen atmosphere unless specifically noted. Flash

chromatography was performed using silica gel (230-400

mesh) from E. M. Science. Proton NMR spectra were recorded

on a General Electric QE300 instrument with Me₄Si as an

internal standard and are reported as shift (multiplicity,

coupling constants, proton counts). Mass spectral analyses

were accomplished using different techniques, including de-

sorption chemical ionization (DCI), atmospheric pressure

chemical ionization (APCI), and electrospray ionization (ESI),

as specified for individual compounds. Elemental analyses

series of compounds against LFA-1/ICAM-1 interaction

and further optimization of the pharmaceutical proper-

ties of this series of compounds will be reported in due

course.

Exp er im en ta l Section

Gen er a l. Unless otherwise specified, all solvents and

reagents were obtained from commercial suppliers and used

without further purification. All reactions were performed

4034 J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 21

Liu et al.

Ta ble 6. Comparison of LFA-1/ICAM-1 Biochemical Assay and

ICAM-1/J Y-8 Cell Adhesion Assay

mmol) was refluxed with morpholine (0.13 mL, 1.55 mmol) in

1.0 mL of MeOH for 16 h. After cooling to room temperature,

the reaction mixture was purified on a preparative HPLC to

give 114 mg of amide 7e (0.28 mmol, 90% yield) as a light

brown oil: ¹H NMR (CDCl₃, 300 MHz) δ 2.45 (br m, 4H), 2.52

(br m, 4H), 3.62 (br m, 4H), 6.53 (d, J ) 8.7 Hz, 1H), 7.05 (d,

J ) 2.4, 8.7 Hz, 1H), 7.17 (td, J ) 0.9, 7.65 Hz, 1H), 7.39 (d,

J ) 2.4 Hz, 1H), 7.46-7.56 (m, 2H), 8.33 (d, J ) 8.7 Hz, 1H);

MS (DCI/NH₃) (M + H)⁺at m/z 411, 413, 415. Anal. (C₁₉H₂₀

Cl₂N₂O₂S‚0.06H₂O) C, H, N.

-

2,4-Dich lor op h en yl 2-(N,N-Dim eth yla m in oeth ylca r bo-

n ylam in o)ph en yl Su lfide (7a): colorless oil; ¹H NMR (CDCl₃,

300 MHz) δ 2.08 (br s, 6H), 2.47 (br m, 2H), 2.75 (br m, 2H),

6.44 (d, J ) 8.7 Hz, 1H), 7.03 (dd, J ) 2.1, 8.4 Hz, 1H), 7.10-

7.23 (m, 2H), 7.39 (br m, 1H), 7.49 (d, J ) 8.4 Hz, 1H), 7.55

(d, J ) 8.4 Hz, 1H); MS (DCI/NH₃) (M + H)⁺at m/z 369, 371,

373. Anal. (C₁₇H₁₈Cl₂N₂OS) C, H, N.

2,4-Dich lor op h en yl 2-(N,N-Di-n -p r op yla m in oeth ylca r -

bon yla m in o)p h en yl Su lfid e (7b): colorless oil; ¹H NMR

(CDCl₃, 300 MHz) δ 0.78 (t, J ) 6.9 Hz, 3H), 1.01 (t, J ) 6.9

Hz, 3H), 1.36 (br m, 2H), 1.53 (br m, 2H), 1.83 (br m, 2H),

2.39 (br m, 2H), 2.67 (br m,1H), 2.94 (br m, 1H), 3.12 (br m,

1H), 3.37 (br m, 1H), 6.44 (d, J ) 8.7 Hz, 1H), 7.03 (dd, J )

2.1, 8.4 Hz, 1H), 7.10-7.23 (m, 2H), 7.39 (br m, 1H), 7.49 (d,

J ) 8.4 Hz, 1H), 7.55 (d, J ) 8.4 Hz, 1H); MS (DCI/NH₃) (M

+ H)⁺at m/z 425, 427, 429. Anal. (C₂₁H₂₆Cl₂N₂OS‚0.04H₂O)

C, H, N.

2,4-Dich lor op h en yl 2-(N-Ben zylm eth yla m in oeth ylca r -

bon yla m in o)p h en yl Su lfid e (7c): colorless oil; ¹H NMR

(CDCl₃, 300 MHz) δ 2.10 (s, 3H), 2.45 (br m, 2H), 2.58 (br m,

2H), 3.56 (s, 2H), 6.47 (d, J ) 8.4 Hz, 1H), 7.02 (dd, J ) 2.7,

8.4 Hz, 1H), 7.11-7.46 (m, 7H), 7.51 (t, J ) 8.4 Hz, 1H), 7.55

(dd, J ) 1.9, 8.4 Hz, 1H), 8.40 (d, J ) 7.5 Hz, 1H); MS (DCI/

NH₃) (M + H)⁺at m/z 445, 447, 449. Anal. (C₂₃H₂₂Cl₂N₂OS‚

0.06H₂O) C, H, N.

a

IC₅₀calculated using a mean of at least two measurements

(all duplicates) for 6 concentrations from 6.4 × 10^-9to 2 × 10^-5

unless otherwise noted.

were performed by Robertson Microlit Laboratories, Madison,

NJ , and are consistent with theoretical values to within 0.4%

unless indicated. Preparative HPLC was performed on an

automated Gilson HPLC system, using an YMC C-18 column,

75 × 30 mm i.d., S-5 µm, 120 Å, and a flow rate of 25 mL/min;

λ ) 214, 245 nm; mobile phase A, 0.05 M NH₄OAc or 0.1%

TFA in H₂O, and mobile phase B, CH₃CN; linear gradient 20-

100% B in 20 min. The purified fractions were evaporated to

dryness on a Savant SpeedVac.

2,4-Dich lor op h en yl 2-(1-P yr r olin yleth ylca r bon yla m i-

1

n o)p h en yl Su lfid e (7d ): colorless oil; H NMR (CDCl₃, 300

MHz) δ 1.66-1.90 (br m, 4H), 2.28-2.63 (br m, 6H), 2.63-

2.83 (br m, 2H), 6.52 (d, J ) 8.7 Hz, 1H), 7.03 (dd, J ) 2.1, 8.7

Hz, 1H), 7.14 (t, J ) 7.2 Hz, 1H), 7.39 (d, J ) 2.4 Hz, 1H),

7.43-7.53 (m, 2H), 8.41 (d, J ) 8.1 Hz, 1H); MS (ESI) (M +

H)⁺at m/z 395, 397, 399. Anal. (C₁₉H₂₀N₂OSCl₂‚0.10H₂O) C,

H, N.

2-[(2,4-Dich lor op h en yl)t h io]n it r ob en zen e (5). To a

stirred solution of 2,4-dichlorothiophenol (2.0 g, 11.2 mmol)

in 25 mL of anhydrous DMF was added potassium carbonate

(3.09 g, 22.4 mmol), followed by 2-chloronitrobenzene (1.78 g,

11.3 mmol). The mixture was then heated under nitrogen

atmosphere at 70 °C for 5 h. The reaction mixture was then

allowed to cool to room temperature and partitioned between

ether and water. The aqueous layer was extracted with ether

once and the combined organic layer was washed with water

and brine, dried over sodium sulfate and condensed in vacuuo

to give 2.78 g (9.25 mmol, 83%) of the desired nitrobenzene as

a light yellow solid.

2-[(2,4-Dich lor oph en yl)th io](2-acr ylam ido)ben zen e (6).

A mixture of compound 5 (1.5 g, 5.0 mmol) and SnCl₂(4.74 g,

25 mmol) in 50 mL of anhydrous EtOH was refluxed under

nitrogen atmosphere for 90 min. The reaction was then allowed

to cool to room temperature, quenched with saturated NaH-

CO₃, extracted with EtOAc (2 × 50 mL). The combined organic

layer was washed with water and brine, dried over Na₂SO₄,

condensed in vacuuo to give 1.29 g of the crude aniline as a

yellowish brown solid. The aniline was taken up in 20 mL of

CH₂Cl₂, Et₃N (1.0 mL, 7.17 mmol) was added, followed by

dropwise addition of acryloyl chloride (0.42 mL, 5.26 mmol).

After stirring for 1 h at ambient temperature, the mixture was

poured into 10 mL of 3 N aqueous HCl and extracted with 20

mL of EtOAc. The organic layer was then washed with brine,

dried over MgSO₄, concentrated under reduced pressure to give

the crude amide which was purified on a silica gel flash column

eluted with 50% EtOAc in hexanes to give 1.05 g (3.24 mmol,

65% over two steps) of the acrylamide as a white solid.

2,4-Dich lor op h en yl 2-(Mor p h olin oeth ylca r bon yla m i-

n o)p h en yl Su lfid e (7e). The acrylamide 6 (100 mg, 0.31

2,4-Dich lor op h en yl 2-(4-Meth yl-1-p ip er a zin yleth ylca r -

bon yla m in o)p h en yl Su lfid e (7f): colorless oil; ¹H NMR

(CDCl₃, 300 MHz) δ 2.28-2.97 (m, 15H), 6.53 (d, J ) 8.7 Hz,

1H), 7.06 (dd, J ) 2.4, 8.7 Hz, 1H), 7.17 (td, J ) 0.9, 7.65 Hz,

1H), 7.41 (d, J ) 2.4 Hz, 1H), 7.46-7.57 (m, 2H), 8.33 (d, J )

8.7 Hz, 1H); MS (DCI/NH₃) (M + H)⁺at m/z 424, 426, 428.

Anal. (C₂₀H₂₃Cl₂N₃OS‚0.17H₂O) C, H, N.

2,4-Dich lor op h en yl 2-(Mor p h olin op r op yla m in o)p h en -

yl Su lfid e (8). To a stirred suspension of LAH (120 mg, 0.29

mmol) in 2.0 mL of anhydrous THF was added amide 7e (22

mg, 0.58 mmol) in 1.0 mL of THF dropwise. After completion

of the addition, the reaction mixture was stirred for 4 h at

room temperature before it was quenched with 22 µL of H₂O,

22 µL of 3 N NaOH, and 66 µL of H₂O. The resulting mixture

was filtered through Celite and condensed in vacuuo. The

resulting residue was purified on a preparative HPLC to give

100 mg (0.25 mmol, 86% yield) of the diamine 8 as a light

1

brown oil: H NMR (CDCl₃, 300 MHz) δ 1.94-2.10 (m, 2H),

2.58-2.81 (br m, 2H), 2.83-2.95 (m, 2H), 3.27 (t, J ) 6.45 Hz,

1H), 3.44 (br d, J ) 8.1 Hz, 2H), 3.87-4.07 (br m, 4H), 6.52

(d, J ) 8.7 Hz, 1H), 6.69 (d, J ) 8.7 Hz, 1H), 6.78 (td, J ) 0.9,

7.5 Hz, 1H), 7.05 (dd, J ) 2.4, 8.7 Hz, 1H), 7.39 (td, J ) 1.8,

7.8 Hz, 2H), 7.49 (d, J ) 1.8, 7.8 Hz, 1H); MS (DCI/NH₃) (M +

H)⁺at m/z 397, 399, 401. Anal. (C₁₉H₂₂N₂OCl₂S‚1.14TFA) C,

H, N.

3-[(2,4-Dich lor op h en yl)th io]ben zoic Acid (12). To a

solution of KOH (1.6 g, 28.5 mmol) in 30 mL of H₂O was added

Cu powder (133 mg, 0.45 mmol), 2,4-dichlorothiophenol (1.5

g, 8.38 mmol) and 3-iodobenzoic acid (2.0 g, 8.38 mmol)

sequentially. The mixture was then refluxed for 9 h before it

was allowed to cool to room temperature. The aqueous mixture

Novel p-Arylthio Cinnamides

J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 21 4035

was then filtered through Celite, washed with hot H₂O,

acidified with concentrated HCl and extracted with CH₂Cl₂(3

× 15 mL). The combined organic layer was washed with brine,

dried over Mg₂SO₄, condensed in vacuuo, and pumped to give

2.44 g (8.16 mmol, 97% yield) of the acid 12 as a white solid:

¹H NMR (DMSO-d₆, 300 MHz) δ 7.15 (d, J ) 8.4 Hz, 1H), 7.42

(dd, J ) 2.1, 8.4 Hz, 1H), 7.58 (t, J ) 7.8 Hz, 1H), 7.66 (dd, J

) 1.8, 7.8 Hz, 1H), 7.78 (d, J ) 2.4 Hz, 1H), 7.85 (d, J ) 2.4

Hz, 1H), 7.94 (dd, J ) 1.8, 7.8 Hz, 1H), 13.2 (s, 1H); MS (DCI/

NH₃) (M + H)⁺at m/z 298, 300.

3-[(2,4-Dich lor op h en yl)th io]ben za ld eh yd e (10m ). To a

stirred solution of acid 12 (1.4 g, 4.68 mmol) in THF was added

1.0 M of BH₃in THF (9.4 mL, 9.36 mmol). The mixture was

then stirred at room temperature for 1 h before it was

quenched carefully with 3 N HCl. The biphasic mixture was

extracted with Et₂O (2 × 25 mL), the combined organic layer

was washed with brine, dried over Na₂SO₄, condensed in

vacuuo to give 1.3 g (4.56 mmol, 97%) of the alcohol as a

colorless oil. To a stirred solution of DMSO (1.74 mL, 24.6

mmol) in 30 mL of CH₂Cl₂at -78 °C was dropped in (COCl)₂

(1.07 mL, 12.3 mmol) slowly. After 20 min, the alcohol (700

mg, 2.46 mmol) in 5 mL of CH₂Cl₂was then added. After 30

min, Et₃N (6.9 mL, 49.2 mmol) was then added and the

resulting mixture was then allowed to warm to room temper-

ature gradually and stirred for 2 h. The mixture was then

partitioned between Et₂O and 3N HCl. The organic layer was

washed with saturated NaHCO₃, brine, dried over Na₂SO₄, and

condensed in vacuuo. The residue was purified on a silica gel

column eluted with 10% Et₂O in hexanes to give 330 mg (1.17

mmol, 47% yield) of the aldehyde 10m as a white solid: ¹H

NMR (CDCl₃, 300 MHz) δ 7.12 (d, J ) 8.4 Hz,1H), 7.18 (dd, J

) 2.1, 8.4 Hz, 1H), 7.47 (d, J ) 2.1 Hz, 1H), 7.53 (d, J ) 8.1

Hz, 1H), 7.58 (td, J ) 1.8, 7.8 Hz, 1H), 7.80 (td, J ) 1.8, 7.8

Hz, 1H), 7.82 (d, J ) 1.8 Hz, 1H), 9.98 (s, 1H); MS (DCI/NH₃)

(M + H)⁺at m/z 282, 284.

2,4-Dich lor op h en yl 2-(N,N-Dim eth yla m in oeth yla m i-

n om eth yl)p h en yl Su lfid e (13). A mixture of the aldehyde

10 (100 mg, 0.35 mmol) prepared analogous to nitrobenzene

5 with N,N-dimethylethylenediamine (38.9 µL, 0.35 mmol) in

2.5 mL of MeOH was stirred at room temperature for 2 h

before NaBH₄(20.8 mg, 0.53 mmol) was added. The resulting

mixture was stirred for an additional 2 h before it was

quenched with saturated NaHCO₃. The reaction mixture was

extracted with CH₂Cl₂(2 × 10 mL), the organic layer was

washed with brine, dried over Mg₂SO₄, condensed in vacuuo.

The resulting residue was purified on a preparative HPLC to

give 117 mg (0.33 mmol, 95% yield) of the diamine 13 as a

colorless oil: ¹H NMR (CDCl₃, 300 MHz) δ 2.00 (s, 6H), 2.42

(t, J ) 6.15 Hz, 2H), 2.70 (t, J ) 6.15 Hz, 2H), 3.89 (s, 2H),

6.65 (d, J ) 8.7 Hz, 1H), 7.04 (dd, J ) 2.4, 8.7 Hz, 1H), 7.30

(dd, J ) 1.5, 7.2 Hz, 1H), 7.37-7.45 (m, 3H), 7.56 (dd, J ) 1.5,

7.2 Hz, 1H); MS (DCI/NH₃) (M + H)⁺at m/z 355, 357, 359.

Anal. (C₁₇H₂₀N₂Cl₂S‚2.13TFA) C, H, N.

tr a n s-2-[(2,4-Dich lor op h en yl)th io]cin n a m ic Acid (14a ).

A mixture of the aldehyde 10 (1.50 g, 5.3 mmol), malonic acid

(1.21 g, 11.6 mmol), and piperidine (78.6 µL, 0.80 mmol) in

8.0 mL of anhydrous pyridine was heated at 110 °C for 2 h

during which the gas evolution ceased. After pyridine was

removed under vacuum, water and 3 N aqueous HCl were then

added with stirring. The desired cinnamic acid was collected

through filtration, washed with cold water and dried in a

vacuum oven overnight to give 1.56 g (4.8 mmol, 91%) of a

white solid: ¹H NMR (DMSO-d₆, 300 MHz) δ 6.57 (d, J ) 15.9

Hz, 1H), 6.69 (d, J ) 8.9 Hz, 1H), 7.34 (d, J ) 2.7, 8.9 Hz,

1H), 7.47-7.60 (m, 3H), 7.74 (d, J ) 2.7 Hz, 1H), 7.89 (d, J )

15.9 Hz, 1H), 8.00 (d, J ) 6.9 Hz, 1H); MS (DCI/NH₃) (M +

H)⁺at m/z 342, 344, 346. Anal. (C₁₅H₁₀O₂Cl₂S‚0.22H₂O) C, H,

N.

1H), 12.5 (s, 1H); MS (DCI/NH₃) (M + H)⁺at m/z 358, 360,

362, 364. Anal. (C₁₅H₉O₂Cl₃S) C, H, N.

2,4-Dich lor op h en yl 2-((E)-((6-H yd r oxyh exyla m in o)-

ca r bon yl)eth en yl)p h en yl Su lfid e (15d ). A suspension of the

acid 14 (284 mg, 0.87 mmol) in 5 mL of methylene chloride

was stirred with (COCl)₂(84 µL, 0.97 mmol), and 1 drop of

DMF under nitrogen atmosphere for 90 min. The solvent was

then removed under vacuum and the residual (COCl)₂was

removed with benzene (2 × 2 mL) in vacuuo. To a separate

flask, previously filled with 6-amino-1-hexanol (12 mg, 0.10

mmol), i-Pr₂NEt (22.8 µL, 0.13 mmol) and DMAP (1.1 mg,

0.008 mmol) in 2.0 mL of CH₂Cl₂, the acid chloride (30 mg,

0.087 mmol) in 1.0 mL of CH₂Cl₂was then dropped in slowly.

After 30 min, the reaction mixture was poured into 3 N

aqueous HCl and extracted with EtOAc. The organic layer was

washed with brine, dried with Na₂SO₄, condensed under

reduced pressure. The crude product was purified on

a

preparative HPLC to give 21.0 mg (0.051 mmol, 90%) of the

title compound as a colorless oil: ¹H NMR (CDCl₃, 300 MHz)

δ 1.31-1.48 (m, 4H), 1.48-1.70 (m, 4H), 3.37 (q, J ) 6.7 Hz,

2H), 3.65 (t, J ) 6.3 Hz, 2H), 5.63 (br s, 1H), 6.36 (d, J ) 15.9

Hz, 1H), 6.71 (d, J ) 9.3 Hz, 1H), 7.05 (dd, J ) 2.4, 8.7 Hz,

1H), 7.31-7.49 (m, 4H), 7.65 (dd, J ) 2.1, 7.5 Hz, 1H), 7.99

(d, J ) 15.9 Hz, 1H); MS (DCI/NH₃) (M + NH₄)⁺at m/z 441,

443, 445. Anal. (C₂₁H₂₃Cl₂NO₂S‚0.03H₂O) C, H, N.

2,4-Dich lor op h en yl 2-((E)-((P h en eth yla m in o)ca r bon -

yl)eth en yl)p h en yl Su lfid e (15a ): white solid; ¹H NMR

(CDCl₃, 300 MHz) δ 2.88 (t, J ) 7.2 Hz, 2H), 3.63 (q, J ) 7.2

Hz, 2H), 5.60 (br s, 1H), 6.32 (d, J ) 15.9 Hz, 1H), 6.70 (d, J

) 8.9 Hz, 1H), 7.04 (dd, J ) 2.7, 8.9 Hz, 1H), 7.18-7.50 (m,

8H), 7.64 (dd, J ) 1.8, 7.5 Hz, 1H), 8.00 (d, J ) 15.9 Hz, 1H);

MS (DCI/NH₃) (M + H)⁺at m/z 428, 430, 432. Anal. (C₂₃H₁₉

Cl₂NOS) C, H, N.

-

2,4-Dich lor op h en yl 2-((E)-((4-Met h ylb en zot h ia zo-2-

yla m in o)ca r bon yl)eth en yl)p h en yl Su lfid e (15b): light

1

brown solid; H NMR (CDCl₃, 300 MHz) δ 2.57 (s, 3H), 6.46

(d, J ) 15.9 Hz, 1H), 6.73 (d, J ) 8.9 Hz, 1H), 7.07 (dd, J )

2.7, 8.9 Hz, 1H), 7.26 (overlapping t, 1H), 7.32-7.51 (m, 5H),

7.70 (dd, J ) 3.3, 5.7 Hz, 1H), 8.35 (d, J ) 15.9 Hz, 1H), 9.95

(br s, 1H); MS (DCI/NH₃) (M + H)⁺at m/z 471, 473, 475. Anal.

(C₂₃H₁₆Cl₂N₂OS₂‚0.07H₂O) C, H, N.

2,4-Dich lor op h en yl 2-((E)-((N-Ben zyl(p-p h en oxyben -

zyl)a m in o)ca r bon yl)eth en yl)p h en yl Su lfid e (15c): white

1

solid; H NMR (CDCl₃, 300 MHz) δ 4.66 (s, 2H), 6.70 (br m,

1H), 6.90-7.16 (m, 4H), 7.06-7.21 (m, 4H), 7.21-7.47 (m, 8H),

7.53 (br m, 1H), 8.22 (br s, 1H); MS (DCI/NH₃) (M + H)⁺at

m/z 597, 599, 601. Anal. (C₃₅H₂₇Cl₂NO₂S‚0.04H₂O) C, H, N.

2,4-Dich lor op h en yl 2-((E)-((2-Hyd r oxyeth yla m in o)ca r -

bon yl)eth en yl)p h en yl Su lfid e (15e): colorless oil; ¹H NMR

(CDCl₃, 300 MHz) δ 3.55 (q, J ) 5.3 Hz, 2H), 3.80 (t, J ) 5.3

Hz, 2H), 6.08 (s, 1H), 6.39 (d, J ) 15.9 Hz, 1H), 6.73 (d, J )

8.9 Hz, 1H), 7.06 (d, J ) 2.7, 8.9 Hz, 1H), 7.32-7.47 (m, 4H),

7.65 (dd, J ) 1.8, 8.9 Hz, 1H), 8.05 (d, J ) 15.9 Hz, 1H); MS

(DCI/NH₃) (M + H)⁺at m/z 368, 370, 372. Anal. (C₁₇H₁₅NO₂-

Cl₂S‚0.33H₂O) C, H, N.

2,4-Dich lor op h en yl 2-((E)-((2,3-Dih yd r oxyp r op yla m i-

n o)ca r bon yl)eth en yl)p h en yl Su lfid e (15f): colorless oil; ¹H

NMR (CDCl₃, 300 MHz) δ 2.94 (br m, 2H), 3.53 (q, J ) 5.5 Hz,

2H), 3.57-3.66 (m, 2H), 3.83 (p, J ) 5.5 Hz, 1H), 6.08 (br m,

1H), 6.38 (d, J ) 15.9 Hz, 1H), 6.75 (d, J ) 8.9 Hz, 1H), 7.06

(dd, J ) 2.7, 8.9 Hz, 1H), 7.32-7.48 (m, 4H), 7.65 (dd, J ) 1.8,

8.9 Hz, 1H), 8.06 (d, J ) 15.9 Hz, 1H); MS (DCI/NH₃) (M +

H)⁺at m/z 398, 401, 403. Anal. (C₁₈H₁₇Cl₂NO₃S‚0.5H₂O) C, H,

N.

2,4-Dich lor op h en yl 2-((E)-((N,N-Dieth a n ola m in o)ca r -

bon yl)eth en yl)p h en yl Su lfid e (15g): colorless oil; ¹H NMR

(CDCl₃, 300 MHz) δ 2.23 (br m, 2H), 3.60 (t, J ) 4.8 Hz, 2H),

3.65 (t, J ) 4.8 Hz, 2H), 3.73 (t, J ) 4.8 Hz, 2H), 4.02 (t, J )

4.8 Hz, 2H), 6.72 (d, J ) 8.9 Hz, 1H), 6.90 (d, J ) 15.9 Hz,

1H), 7.05 (dd, J ) 2.7, 8.9 Hz, 1H), 7.32-7.49 (m, 4H), 7.67

(d, J ) 8.9 Hz, 1H), 8.08 (d, J ) 15.9 Hz, 1H); MS (DCI/NH₃)

(M + H)⁺at m/z 412, 414, 416. Anal. (C₁₉H₁₉NO₃Cl₂S‚0.24H₂O)

C, H, N.

tr a n s-2-Ch lor o-4-[(2,4-d ich lor op h en yl)t h io]cin n a m ic

Acid (14b): white solid; ¹H NMR (DMSO-d₆, 300 MHz) δ 6.62

(d, J ) 15.9 Hz, 1H), 7.05 (d, J ) 8.9 Hz, 1H), 7.35 (d, J ) 8.9

Hz, 1H), 7.49 (dd, J ) 2.7, 8.9 Hz, 1H), 7.55 (d, J ) 15.9 Hz,

1H), 7.64 (d, J ) 8.9 Hz, 1H), 7.85 (d, J ) 2.7 Hz, 1H), 7.98 (s,

4036 J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 21

Liu et al.

2,4-Dich lor op h en yl 2-((E)-(Glycin ylca r bon yl)eth en yl)-

p h en yl Su lfid e (15h ): white solid; ¹H NMR (CDCl₃, 300 MHz)

δ 4.13 (br m, 1H), 4.22 (d, J ) 4.8 Hz, 2H), 6.29 (t, J ) 4.65

Hz, 1H), 6.46 (d, J ) 15.9 Hz, 1H), 6.74 (d, J ) 8.9 Hz, 1H),

7.06 (dd, J ) 2.7, 8.9 Hz, 1H), 7.33-7.47 (m, 4H), 7.67 (dd, J

) 1.8, 8.9 Hz, 1H), 8.10 (d, J ) 15.9 Hz, 1H); MS (DCI/NH₃)

(M + NH₄)⁺at m/z 399, 401, 403. Anal. (C₁₇H₁₃NO₃Cl₂S‚

0.09TFA) C, H, N.

2,4-Dich lor op h en yl 2-((E)-((3-(1-Im id a zolyl)p r op yla m i-

n o)ca r bon yl)eth en yl)p h en yl Su lfid e (15i): white powder;

¹H NMR (DMSO-d₆, 300 MHz) δ 1.88 (p, J ) 7.7 Hz, 2H), 3.11

(q, J ) 7.7 Hz, 2H), 3.97 (t, J ) 7.7 Hz, 2H), 6.63 (d, J ) 15.9

Hz, 1H), 6.70 (d, J ) 8.7 Hz, 1H), 6.89 (d, J ) 0.9 Hz, 1H),

7.17 (d, J ) 0.9 Hz, 1H), 7.33 (dd, J ) 2.7, 8.7 Hz, 1H), 7.46-

7.65 (m, 4H), 7.72 (d, J ) 2.7 Hz, 1H), 7.78 (d, J ) 15.9 Hz,

1H), 7.80 (d, J ) 8.7 Hz, 1H), 8.24 (t, J ) 5.9 Hz, 1H); MS

(DCI/NH₃) (M + H)⁺at m/z 448, 450, 452. Anal. (C₂₁H₁₉N₃-

OCl₂S‚0.87H₂O) C, H, N.

(DMSO-d₆, 300 MHz) δ 3.51-3.67 (m, 6H), 3.67-3.77 (m, 2H),

6.94 (d, J ) 8.9 Hz, 1H), 7.35 (d, J ) 15.3 Hz, 1H), 7.37 (dd,

J ) 2.7, 8.9 Hz, 1H), 7.40 (d, J ) 15.3 Hz, 1H), 7.48 (s, 1H),

7.52 (d, J ) 8.9 Hz, 1H), 7.74 (d, J ) 2.7 Hz, 1H), 7.79 (d, J )

8.9 Hz, 1H), 7.98 (s, 1H); MS (DCI/NH₃) (M + H)⁺at m/z 394,

396, 398. Anal. (C₁₉H₁₇NO₂Cl₂S) C, H, N.

2,4-Dich lor op h en yl 3-((E)-((3-(1-P yr r olid in -2-on yl)p r o-

p yla m in o)ca r bon yl)eth en yl)p h en yl Su lfid e (16e): color-

1

less oil; H NMR (CDCl₃, 300 MHz) δ 1.74 (quintet, J ) 5.9

Hz, 2H), 2.08 (quintet, J ) 7.6 Hz, 2H), 2.44 (t, J ) 7.6 Hz,

2H), 3.33 (q, J ) 5.9 Hz, 2H), 3.41 (q, J ) 6.7 Hz, 4H), 3.33 (q,

J ) 5.9 Hz, 2H), 3.41 (q, J ) 6.7 Hz, 4H), 6.45 (d, J ) 15.3 Hz,

1H), 6.95 (d, J ) 8.9 Hz, 1H), 7.12 (dd, J ) 2.7, 8.9 Hz, 1H),

7.30-7.39 (m, 2H), 7.43 (d, J ) 2.7 Hz, 1H), 7.50 (td, J ) 1.8,

6.6 Hz, 1H), 7.56 (d, J ) 15.3 Hz, 1H), 7.56 (s, 1H); MS (DCI/

NH₃) (M + H)⁺at m/z 449, 451, 453. Anal. (C₂₂H₂₂N₂O₂Cl₂S‚

0.25H₂O) C, H, N.

2,4-Dich lor op h en yl 2-Ch lor o-4-((E)-((2-h yd r oxyeth yl-

a m in o)ca r bon yl)eth en yl)p h en yl Su lfid e (17a ): colorless

2,4-Dich lor op h en yl 2-((E)-((2-Tetr a h yd r ofu r ylm eth yl-

1

a m in o)ca r bon yl)eth en yl)p h en yl Su lfid e (15j): white pow-

oil; H NMR (CDCl₃, 300 MHz) δ 3.57 (q, J ) 7.65 Hz, 2H),

1

der; H NMR (CDCl₃, 300 MHz) δ 1.91 (p, J ) 7.05 Hz, 2H),

3.71 (q, J ) 7.65 Hz, 2H), 6.06 (br s, 1H), 6.40 (d, J ) 15.3 Hz,

1H), 6.96 (d, J ) 8.7 Hz, 1H), 7.22-7.30 (m, 4H), 7.49-7.60

(m, 1H), 7.55 (d, J ) 15.3 Hz, 1H); MS (APCI) (M + H)⁺at

m/z 402, 404, 406, 408. Anal. (C₁₇H₁₄NO₂Cl₃S‚0.25H₂O) C, H,

N.

1.50-1.66 (m, 2H), 3.18-3.31 (m, 1H), 3.63-3.93 (m, 3H), 4.01

(qd, J ) 3.0, 6.9 Hz, 1H), 5.98 (m, 1H), 6.38 (d, J ) 15.9 Hz,

1H), 6.70 (d, J ) 8.9 Hz, 1H), 7.04 (d, J ) 2.7, 8.9 Hz, 1H),

7.32-7.49 (m, 4H), 7.66 (d, J ) 1.8, 8.9 Hz, 1H), 8.03 (d, J )

15.9 Hz, 1H); MS (DCI/NH₃) (M + H)⁺at m/z 408, 410, 412.

Anal. (C₂₀H₁₉NO₂Cl₂S‚0.50H₂O) C, H, N.

2,4-Dich lor op h en yl 2-Ch lor o-4-((E)-((6-h yd r oxyh exyl-

a m in o)ca r bon yl)eth en yl)p h en yl Su lfid e (17b): colorless

2,4-Dich lor op h en yl 2-((E)-((3-(1-P yr r olid in -2-on yl)p r o-

1

oil; H NMR (CDCl₃, 300 MHz) δ 1.42 (m, 4H), 1.58 (m, 4H),

p yla m in o)ca r bon yl)eth en yl)p h en yl Su lfid e (15l): colorless

3.40 (q, J ) 6.7 Hz, 2H), 3.65 (br m, 2H), 5.60 (br t, 1H), 6.35

(d, J ) 15.3 Hz, 1H), 6.98 (d, J ) 8.7 Hz, 1H), 7.22-7.30 (m,

4H), 7.49-7.60 (m, 1H), 7.55 (d, J ) 15.3 Hz, 1H); MS (APCI)

(M + H)⁺at m/z 458, 460, 462, 464. Anal. (C₂₁H₂₂NO₂Cl₃S‚

0.27H₂O) C, H, N.

1

oil; H NMR (CDCl₃, 300 MHz) δ 1.73 (quintet, J ) 6.1 Hz,

2H), 2.07 (quintet, J ) 7.8 Hz, 2H), 2.44 (t, J ) 7.8 Hz, 2H),

3.30 (q, J ) 6.1 Hz, 2H), 3.34-3.48 (m, 4H), 6.44 (d, J ) 15.9

Hz, 1H), 6.67 (d, J ) 8.9 Hz, 1H), 7.03 (dd, J ) 2.7, 8.9 Hz,

1H), 7.07 (br t, 1H), 7.30-7.49 (m, 4H), 7.71 (dd, J ) 1.8, 8.9

Hz, 1H), 8.06 (d, J ) 15.9 Hz, 1H); MS (DCI/NH₃) (M + H)⁺

at m/z 449, 451, 453. Anal. (C₂₂H₂₂Cl₂N₂O₂S‚0.23H₂O) C, H,

N.

2,4-Dich lor op h en yl 2-Ch lor o-4-((E)-((b is(2-h yd r oxy-

eth yl)a m in o)ca r bon yl)eth en yl)p h en yl Su lfid e (17c): col-

1

orless oil; H NMR (CDCl₃, 300 MHz) δ 2.99 (br s, 2H), 3.67

(br m, 4H), 3.88 (t, J ) 5.1 Hz, 2H), 3.94 (t, J ) 5.1 Hz, 2H),

6.94 (d, J ) 15.3 Hz, 1H), 6.97 (d, J ) 8.7 Hz, 1H), 7.21-7.32

(m, 3H), 7.50-7.54 (m, 1H), 7.58 (d, J ) 2.4 Hz, 1H), 7.58 (d,

J ) 15.3 Hz, 1H); MS (APCI) (M + H)⁺at m/z 446, 448, 450,

452. Anal. (C₁₉H₁₈NO₃Cl₃S‚1.09H₂O) C, H, N.

2,4-Dich lor op h en yl 2-((E)-((1-Mor p h olin o)ca r bon yl)-

eth en yl)p h en yl Su lfid e (15m ): white powder; ¹H NMR

(CDCl₃, 300 MHz) δ 3.53 (br m, 2H), 3.55-3.62 (m, 4H), 3.67

(br m, 2H), 6.66 (d, J ) 8.9 Hz, 1H), 7.28 (d, J ) 15.9 Hz, 1H),

7.33 (dd, J ) 2.7, 8.9 Hz, 1H), 7.42-7.62 (m, 3H), 7.74 (d, J )

2.7 Hz, 1H), 7.85 (d, J ) 15.9 Hz, 1H), 8.11 (d, J ) 7.8 Hz,

2,4-Dich lor op h en yl 2-Ch lor o-4-((E)-((3-(1-im id a zolyl)-

pr opylam in o)car bon yl)eth en yl)ph en yl Su lfide (17d): white

1H); MS (APCI) (M + H)⁺at m/z 394, 396, 398. Anal. (C₁₉H₁₇

Cl₂NO₂S‚0.09H₂O) C, H, N.

-

1

powder; H NMR (CDCl₃, 300 MHz) δ 2.09 (p, J ) 6.75 Hz,

2H), 3.41 (q, J ) 6.75 Hz, 2H), 4.05 (t, J ) 6.75 Hz, 2H), 5.81

(br m, 1H), 6.33 (d, J ) 15.9 Hz, 1H), 6.95 (d, J ) 8.9 Hz, 1H),

6.98 (br s, 1H), 7.10 (br s, 1H), 7.24 (s, 2H), 7.27 (overlapping

m, 2H), 7.52 (m, 1H), 7.52 (d, J ) 15.9 Hz, 1H), 7.56 (d, J )

2.1 Hz, 1H); MS (APCI) (M + H)⁺at m/z 466, 468, 470. Anal.

(C₂₁H₁₈N₃OCl₃S‚0.71H₂O) C, H, N.

2,4-Dich lor op h en yl 3-((E)-((6-H yd r oxyh exyla m in o)-

ca r bon yl)eth en yl)p h en yl Su lfid e (16a ): colorless oil; ¹H

NMR (CDCl₃, 300 MHz) δ 1.31-1.48 (m, 4H), 1.48-1.72 (m,

4H), 3.40 (q, J ) 6.9 Hz, 2H), 3.65 (t, J ) 6.9 Hz, 2H), 5.61 (br

t, 1H), 6.36 (d, J ) 15.3 Hz, 1H), 6.98 (d, J ) 8.7 Hz, 1H), 7.13

(dd, J ) 2.4, 8.7 Hz, 1H), 7.33-7.50 (m, 4H), 7.53 (s, 1H), 7.58

(d, J ) 15.3 Hz, 1H); MS (DCI/NH₃) (M + H)⁺at m/z 424, 426,

428. Anal. (C₂₁H₂₃NO₂Cl₂S‚0.05H₂O) C, H, N.

2,4-Dich lor op h en yl 3-((E)-((N,N-Dieth a n ola m in o)ca r -

bon yl)eth en yl)p h en yl Su lfid e (16b): colorless oil; ¹H NMR

(CDCl₃, 300 MHz) δ 3.61-3.74 (m, 4H), 3.88 (t, J ) 4.8 Hz,

2H), 3.94 (t, J ) 4.8 Hz, 2H), 6.93 (d, J ) 15.3 Hz, 1H), 6.97

(d, J ) 8.7 Hz, 1H), 7.13 (dd, J ) 2.4, 8.7 Hz, 1H), 7.33-7.40

(m, 2H), 7.44 (d, J ) 2.4 Hz, 1H), 7.48 (td, J ) 1.8, 6.6 Hz,

1H), 7.53 (s, 1H), 7.63 (d, J ) 15.3 Hz, 1H); MS (DCI/NH₃) (M

+ H)⁺at m/z 412, 414, 416. Anal. (C₁₉H₁₉NO₃Cl₂S‚0.10H₂O)

C, H, N.

2,4-Dich lor op h en yl 3-((E)-((3-(1-Im id a zolyl)p r op yla m i-

n o)ca r bon yl)eth en yl)p h en yl Su lfid e (16c): white powder;

¹H NMR (CDCl₃, 300 MHz) δ 2.02-2.34 (m, 2H), 3.39 (q, J )

6.1 Hz, 2H), 4.15 (t, J ) 6.1 Hz, 2H), 6.50 (d, J ) 15.3 Hz,

1H), 6.57 (br s, 1H), 6.95 (d, J ) 8.7 Hz, 1H), 7.03 (s, 1H), 7.12

(dd, J ) 2.4, 8.7 Hz, 1H), 7.15 (s, 1H), 7.31-7.39 (m, 2H), 7.43

(d, J ) 2.4 Hz, 1H), 7.46 (td, J ) 1.8, 6.6 Hz, 1H), 7.52 (s, 1H),

7.58 (d, J ) 15.3 Hz, 1H), 8.28 (s, 1H); MS (DCI/NH₃) (M +

H)⁺at m/z 432, 434, 436. Anal. (C₂₁H₁₉N₃OCl₂S‚0.50H₂O) C,

H, N.

2,4-Dich lor op h en yl 2-Ch lor o-4-((E)-((3-(1-p yr r olid in -2-

on yl)pr opylam in o)car bon yl)eth en yl)ph en yl Su lfide (17e):

1

colorless oil; H NMR (CDCl₃, 300 MHz) δ 1.74 (quintet, J )

6.0 Hz, 2H), 2.09 (quintet, J ) 7.5 Hz, 2H), 2.45 (t, J ) 8.25

Hz, 2H), 3.33 (q, J ) 6.0 Hz, 2H), 3.42 (q, J ) 8.25 Hz, 4H),

6.46 (d, J ) 15.6 Hz, 1H), 7.02 (d, J ) 8.7 Hz, 1H), 7.14-7.23

(m, 2H), 7.30 (dd, J ) 2.4, 8.7 Hz, 1H), 7.51 (d, J ) 2.4 Hz,

1H), 7.51 (d, J ) 15.6 Hz, 1H), 7.60 (d, J ) 2.1 Hz, 1H); MS

(DCI/NH₃) (M + H)⁺at m/z 483, 485, 487, 489. Anal.

(C₂₂H₂₁N₂O₂Cl₃S‚0.57H₂O) C, H, N.

2,4-Dich lor op h en yl 2-Ch lor o-4-((E)-((1-m or p h olin o)-

ca r bon yl)eth en yl)p h en yl Su lfid e (17f): white solid; ¹H

NMR (CDCl₃, 300 MHz) δ 3.59-3.80 (m, 8H), 6.83 (d, J ) 15.6

Hz, 1H), 6.97 (d, J ) 8.7 Hz, 1H), 7.16-7.32 (m, 3H), 7.49-

7.53 (m, 1H), 7.59 (d, J ) 2.4 Hz, 1H), 7.59 (d, J ) 15.6 Hz,

1H); MS (DCI/NH₃) (M + H)⁺at m/z 428, 430, 432, 434. Anal.

(C₁₉H₁₆NO₂Cl₃S‚0.46H₂O) C, H, N.

2,4-Dich lor oph en yl 2-Ch lor o-4-((E)-((4-m eth ylpiper azin -

1-yl)ca r bon yl)eth en yl)p h en yl Su lfid e (17g): colorless oil;

¹H NMR (CDCl₃, 300 MHz) δ 2.37 (s, 3H), 2.51 (br m, 4H),

3.63-3.87 (br m, 4H), 6.85 (d, J ) 15.6 Hz, 1H), 6.98 (d, J )

8.7 Hz, 1H), 7.19-7.25 (m, 2H), 7.27 (dd, J ) 2.1, 8.7 Hz, 1H),

7.52 (t, J ) 0.9 Hz, 1H), 7.57 (d, J ) 15.6 Hz, 1H), 7.60 (d, J

2,4-Dich lor op h en yl 3-((E)-((1-Mor p h olin o)ca r bon yl)-

eth en yl)p h en yl Su lfid e (16d ): white powder; ¹H NMR

Novel p-Arylthio Cinnamides

J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 21 4037

) 2.1 Hz, 1H); MS (DCI/NH₃) (M + H)⁺at m/z 441, 443, 445,

447. Anal. (C₂₀H₁₉N₂OCl₃S‚0.45H₂O) C, H, N.

Na₂SO₄, concentrated on a rotavap. The crude bromide (230

mg) was used directly for the alkylation without purification.

2,4-Dich lor oph en yl 2-Ch lor o-4-((E)-((4-acetylpiper azin -

1-yl)ca r bon yl)eth en yl)p h en yl Su lfid e (17h ): white solid;

¹H NMR (CDCl₃, 300 MHz) δ 2.15 (s, 3H), 3.50-3.58 (m, 2H),

3.58-3.85 (m, 6H), 6.85 (d, J ) 15.3 Hz, 1H), 6.96 (d, J ) 8.7

Hz, 1H), 7.24-7.36 (m, 3H), 7.54 (d, J ) 2.4 Hz, 1H), 7.61 (d,

J ) 15.3 Hz, 1H), 7.61 (d, J ) 2.1 Hz, 1H); MS (DCI/NH₃) (M

+ H)⁺at m/z 486, 488, 490, 492. Anal. (C₂₁H₁₉N₂O₂Cl₃S‚

0.85H₂O) C, H, N.

To a stirred solution of morpholine (10 µL, 0.11 mmol) in

0.5 mL of CH₃CN was added i-Pr₂NEt (23.7 µL, 0.14 mmol),

followed by the bromide (40 mg, 0.091 mmol). The mixture

was then stirred at room temperature for 2 h. Solvent was

then removed and the crude product was purified with a

preparative HPLC to give 36.3 mg (0.079 mmol, 87% yield) of

compound 21a as a white solid: ¹H NMR (DMSO-d₆, 300 MHz)

δ 2.33 (br t, 4H), 3.45 (br t, 4H), 3.50-3.65 (m, 6H), 3.56 (s,

2H), 3.65-3.80 (br m, 2H), 6.74 (d, J ) 8.7 Hz, 1H), 7.30 (d, J

) 15.3 Hz, 1H), 7.35-7.41 (m, 2H), 7.43 (d, J ) 15.3 Hz, 1H),

7.46 (td, J ) 2.4, 8.1 Hz, 1H), 7.52 (dd, J ) 2.1, 8.7 Hz, 1H),

7.56 (d, J ) 8.1 Hz, 1H), 8.02 (d, J ) 2.1 Hz, 1H); MS (DCI/

NH₃) (M + H)⁺at m/z 459, 461. Anal. (C₂₄H₂₇ClN₂O₃S‚

1.12TFA) C, H, N.

2-(4-F or m ylp ip er a zin -1-ylm eth yl)p h en yl 2-Ch lor o-4-

((E)-((1-m or p h olin o)ca r b on yl)et h en yl)p h en yl Su lfid e

(21b): white solid; ¹H NMR (DMSO-d₆, 300 MHz) δ 2.20-2.32

(m, 6H), 2.74 (br m, 2H), 3.48 (s, 2H), 3.59 (m, 6H), 3.70 (br

m, 2H), 6.74 (d, J ) 8.7 Hz, 1H), 7.29 (d, J ) 15.6 Hz, 1H),

7.35-7.41 (m, 2H), 7.42 (d, J ) 15.6 Hz, 1H), 7.45-7.52 (m,

3H), 7.98 (d, J ) 2.1, 1H); MS (DCI/NH₃) (M + H)⁺at m/z

486, 488. Anal. (C₂₅H₂₈ClN₃O₃S‚0.34TFA) C, H, N.

2-(4-(1,3-Ben zod ioxol-5-ylm eth yl)p ip er a zin -1-ylm eth -

yl)ph en yl 2-Ch lor o-4-((E)-((1-m or ph olin o)car bon yl)eth en -

yl)p h en yl Su lfid e (21c): white solid; ¹H NMR (DMSO-d₆,

300 MHz) δ 2.13-2.40 (br m, 8H), 3.28 (s, 2H), 3.49-3.64 (br

m, 6H), 3.54 (s, 2H), 3.70 (br m, 2H), 5.97 (s, 2H), 6.69 (dd, J

) 1.8, 8.1 Hz, 1H), 6.74 (d, J ) 8.7 Hz, 1H), 6.79 (d, J ) 1.8

Hz, 1H), 6.81 (d, J ) 8.1 Hz, 1H), 7.39 (d, J ) 15.3 Hz, 1H),

7.33-7.38 (m, 2H), 7.38-7.50 (m, 2H), 7.43 (d, J ) 15.3 Hz,

1H), 7.53 (d, J ) 8.4 Hz, 1H), 8.00 (d, J ) 2.1 Hz, 1H); MS

(DCI/NH₃) (M + H)⁺at m/z 592, 594. Anal. (C₃₂H₃₄ClN₃O₄S‚

1.07TFA) C, H, N.

2,4-Dich lor op h en yl 2-Ch lor o-4-((E)-((4-(2-p yr id yl)p ip -

er a zin -1-yl)ca r bon yl)eth en yl)p h en yl Su lfid e (17i): white

1

solid; H NMR (CDCl₃, 300 MHz) δ 3.59 (br m, 2H), 3.69 (br

m, 2H), 3.78 (br m, 2H), 3.86 (br m, 2H), 6.64-6.72 (m, 2H),

6.90 (d, J ) 15.6 Hz, 1H), 6.99 (d, J ) 8.7 Hz, 1H), 7.22-7.25

(m, 2H), 7.31(dd, J ) 2.4, 8.7 Hz, 1H), 7.49-7.57 (m, 2H), 7.61

(d, J ) 15.6 Hz, 1H), 7.62 (d, J ) 2.4 Hz, 1H), 8.19-8.24 (m,

1H); MS (DCI/NH₃) (M + H)⁺at m/z 504, 506, 508, 510. Anal.

(C₂₄H₂₀N₃OCl₃S) C, H, N.

2-Br om oph en yl 2-Ch lor o-4-((E)-((1-m or ph olin o)car bon -

yl)eth en yl)p h en yl Su lfid e (18). Prepared analogously to

compound 15d , substituting 2,4-dichlorothiophenol with 2-bro-

mothiophenol, 2-chlorobenzaldehyde with 3-chloro-4-fluoro-

benzadehyde, and 6-amino-1-hexanol with morpholine: white

1

solid; H NMR (DMSO-d₆, 300 MHz) δ 3.50-3.66 (br m, 6H),

3.66-3.79 (br m, 2H), 7.05 (d, J ) 8.7 Hz, 1H), 7.26 (dd, J )

2.1, 8.1 Hz, 1H), 7.33 (dd, J ) 2.1, 8.1 Hz, 1H), 7.36 (d, J )

15.6 Hz, 1H), 7.39 (dd, J ) 1.8, 12.0 Hz, 1H), 7.45 (dd, J )

1.8, 6.3 Hz, 1H), 7.48 (d, J ) 15.6 Hz,1H), 7.64 (dd, J ) 2.1,

8.7 Hz, 1H), 7.80 (dd, J ) 2.8, 8.7 Hz, 1H), 8.09 (d, J ) 2.1 Hz,

1H); MS (DCI/NH₃) (M + H)⁺at m/z 438, 440, 442.

2-((E)-((1-Mor ph olin o)car bon yl)eth en yl)ph en yl 2-Ch lo-

r o-4-((E)-((1-m or p h olin o)ca r bon yl)eth en yl)p h en yl Su l-

fid e (19). A mixture of bromide 18 (80 mg, 0.18 mmol),

acryloylmorpholine (33 mg, 0.23 mmol), Pd(OAc)₂(2.0 mg,

0.009 mmol), P(o-tolyl)₃(17 mg, 0.056 mmol), Et₃N (39 µL, 0.27

mmol), and anhydrous DMF (1.0 mL) in a pressure tube was

flushed with nitrogen for 5 min before it capped and heated

at 110 °C overnight. TLC indicated almost complete consump-

tion of the starting bromide. The reaction mixture was then

allowed to cool to room temperature, partitioned between

EtOAc and water. The aqueous layer was extracted once with

EtOAc. The combined organic layer was washed with water

and brine, dried over Na₂SO₄, condensed under reduced

pressure. The crude product was purified on a preparative

HPLC to give 35 mg (0.070 mmol, 39%) of compound 19 as a

light brown solid: ¹H NMR (DMSO-d₆, 300 MHz) δ 3.43-3.88

(m, 16H), 6.58 (d, J ) 8.7 Hz, 1H), 7.30 (d, J ) 15.3 Hz, 2H),

7.43 (d, J ) 15.3 Hz, 1H), 7.47-7.64 (m, 4H), 7.86 (d, J ) 15.3

Hz, 1H), 8.06 (d, J ) 2.1 Hz, 1H), 8.14 (d, J ) 7.5 Hz, 1H); MS

(DCI/NH₃) (M + NH₄)⁺at m/z 516, 518. Anal. (C₂₆H₂₇N₂O₄-

ClS‚0.46H₂O) C, H, N.

2-(Hydr oxym eth yl)ph en yl 2-Ch lor o-4-((E)-((1-m or ph oli-

n o)ca r bon yl)eth en yl)p h en yl Su lfid e (20). Prepared analo-

gously to compound 18, substituting 2,4-dichlorothiophenol

with 2-mercaptobenzyl alcohol: white solid; ¹H NMR (CDCl₃,

300 MHz) δ 3.50-3.62 (br m, 6H), 3.65-3.74 (br m, 2H), 4.54

(d, J ) 5.7 Hz, 2H), 5.33 (t, J ) 5.7 Hz, 1H), 6.62 (d, J ) 8.7

Hz, 1H), 7.28 (d, J ) 15.0 Hz, 1H), 7.36 (d, J ) 7.8 Hz, 1H),

7.42 (d, J ) 15.0 Hz, 1H), 7.43 (dd, J ) 1.8, 8.7 Hz, 1H), 7.50

(dd, J ) 2.1, 8.7 Hz, 1H), 7.55 (dd, J ) 2.1, 7.8 Hz, 1H), 7.68

(dd, J ) 1.5, 8.1 Hz, 1H), 8.02 (d, J ) 2.1 Hz, 1H); MS (DCI/

NH₃) (M + H)⁺at m/z 390, 392.

2-((N-Eth oxyca r bon ylm eth yl-N-m eth yla m in o)m eth yl)-

ph en yl 2-Ch lor o-4-((E)-((1-m or ph olin o)car bon yl)eth en yl)-

1

p h en yl Su lfid e (21d ): white solid; H NMR (DMSO-d₆, 300

MHz) δ 1.16 (t, J ) 7.2 Hz, 3H), 2.27 (s, 2H), 3.30 (s, 2H),

3.51-3.66 (br m, 6H), 3.66-3.75 (br m, 2H), 3.78 (s, 2H), 4.05

(q, J ) 7.2 Hz, 2H), 6.75 (d, J ) 8.7 Hz, 1H), 7.30 (d, J ) 15.3

Hz, 1H), 7.33-7.38 (m, 2H), 7.42-7.50 (m, 2H), 7.43 (d, J )

15.3 Hz, 1H), 7.53 (dd, J ) 2.1, 8.7 Hz, 1H), 7.60 (d, J ) 7.8

Hz, 1H), 8.02 (d, J ) 2.1 Hz, 1H); MS (DCI/NH₃) (M + H)⁺at

m/z 489, 491. Anal. (C₂₅H₂₉ClN₂O₄S‚0.42TFA) C, H, N.

2-((3-(1-Mor p h olin o)p r op yl)a m in o)p h en yl 2-Ch lor o-4-

((E)-((1-m or ph olin o)car bon yl)eth en yl)ph en yl Su lfide (22).

A mixture of bromide 18 (60 mg, 0.14 mmol), aminopropyl-

morpholine (24 µL, 0.17 mmol), Pd₂(dba)₃(1.2 mg, 0.0013

mmol), BINAP (2.5 mg, 0.004 mmol), NaOt-Bu (19 mg, 0.20

mmol), 18-crown-6 (50 mg, 0.20 mmol), and anhydrous toluene

(1 mL) in a pressure tube was flushed with nitrogen for 3 min

before it was capped and heated at 80 °C overnight. The

reaction was then stopped and allowed to cool to room

temperature. The reaction mixture was partitioned between

EtOAc and water, and the aqueous layer was extracted once

with EtOAc. The combined organic layer was then washed

with water and brine, dried over Na₂SO₄, condensed under

reduced pressure. The crude product was purified with a

preparative HPLC to give 30 mg (0.062 mmol, 44%) of

compound 22 as a light brown oil: ¹H NMR (DMSO-d₆, 300

MHz) δ 1.62 (quintet, J ) 6.5 Hz, 2H), 2.15-2.26 (m, 8H), 3.17

(q, J ) 6.5 Hz, 2H), 3.22-3.76 (m, 12 H), 3.50 (t, J ) 6.5 Hz,

2H), 5.72 (t, J ) 5.7 Hz, 1H), 6.47 (d, J ) 8.7 Hz, 1H), 6.68 (t,

J ) 7.2 Hz, 1H), 6.81 (d, J ) 8.4 Hz, 1H), 7.26 (d, J ) 15.6 Hz,

1H), 7.35-7.42 (m, 2H), 7.43 (d, J ) 15.6 Hz, 1H), 7.44 (d, J

) 8.4 Hz, 1H), 7.49 (d, J ) 8.4 Hz, 1H), 8.00 (d, J ) 2.1 Hz,

2-(1-Mor ph olin om eth yl)ph en yl 2-Ch lor o-4-((E)-((1-m or -

p h olin o)ca r bon yl)et h en yl)p h en yl Su lfid e (21a ). To a

stirred solution of benzyl alcohol 20 (195 mg, 0.32 mmol) in

2.0 mL of anhydrous DMF was added LiBr (48 mg, 0.35 mmol).

The mixture was then cooled in an ice-water bath, and PBr₃

(60 µL, 0.40 mmol) was dropped in slowly. The ice bath was

then removed and the mixture was stirred at room tempera-

ture for 1 h. Water was then added, the mixture was then

partitioned between EtOAc and aqueous NaHCO₃and the

aqueous layer was extracted with EtOAc once. The combined

organic layer was washed with water and brine, dried over

1H); MS (APCI) (M + H)⁺at m/z 502, 504. Anal. (C₂₆H₃₂

ClN₃O₃S‚1.8TFA) C, H, N.

-

3-Br om o-4-[(2,4-dich lor oph en yl)th io]ben zaldeh yde (23).

Prepared analogously to compound 5, substituting 2-chloroni-

trobenzene with 3-bromo-4-fluorobenzaldehyde: white solid;

¹H NMR (DMSO-d₆, 300 MHz) δ 7.23 (d, J ) 2.7 Hz, 1H), 7.27

(d, J ) 2.7 Hz, 1H), 7.43 (dd, J ) 2.7, 8.4 Hz, 1H), 7.80 (dd, J

4038 J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 21

Liu et al.

) 2.1, 8.4 Hz, 1H), 8.42 (d, J ) 2.1 Hz, 1H), 10.01 (s, 1H); MS

(DCI/NH₃) (M + H)⁺at m/z 360, 362, 364.

Hz, 1H), 7.44 (d, J ) 15.3 Hz, 1H), 7.55 (d, J ) 15.3 Hz, 1H),

7.63 (dd, J ) 2.7, 8.7 Hz, 1H), 7.83 (d, J ) 8.7 Hz, 1H), 7.93

(d, J ) 8.7 Hz, 1H), 7.96 (d, J ) 2.7 Hz, 1H), 8.69 (d, J ) 1.8

Hz, 1H); MS (DCI/NH₃) (M + NH₄)⁺at m/z 497, 499, 501. Anal.

(C₂₁H₁₉N₃O₄Cl₂S‚0.82H₂O) C, H, N.

2,4-Dich lor op h en yl 2-((E)-((1-Mor p h olin o)ca r bon yl)-

eth en yl)-5-for m ylp h en yl Su lfid e (24). Prepared analo-

gously to compound 19, substituting bromide 18 with bromide

23 as the starting material: white solid; ¹H NMR (DMSO-d₆,

300 MHz) δ 3.52-3.65 (m, 6H), 3.68-3.77 (m, 2H), 7.28 (d, J

) 2.7 Hz, 1H), 7.31 (d, J ) 2.7 Hz, 1H), 7.42 (d, J ) 15.3 Hz,

1H), 7.47 (dd, J ) 2.7, 8.4 Hz, 1H), 7.84 (dd, J ) 2.1, 8.4 Hz,

1H), 7.88 (d, J ) 15.3 Hz, 1H), 8.48 (d, J ) 2.1 Hz, 1H), 10.03

(s, 1H); MS (DCI/NH₃) (M + H)⁺at m/z 422, 424, 426.

2,4-Dich lor op h en yl 2-Nit r o-4-((E)-((3-(1-p yr r olid in -2-

on yl)p r op yla m in o)ca r b on yl)et h en yl)p h en yl

Su lfid e

1

(28a ): light yellow powder; H NMR (DMSO-d₆, 300 MHz) δ

1.64 (quintet, J ) 7.1 Hz, 2H), 1.91 (quintet, J ) 7.5 Hz, 2H),

2.21 (t, J ) 8.3 Hz, 2H), 3.15 (q, J ) 6.3 Hz, 2H), 3.21 (dd, J

) 9.9, 17.7 Hz, 2H), 3.32 (overlapping t, J ) 8.4 Hz, 2H), 6.72

(d, J ) 15.6 Hz, 1H), 6.86 (d, J ) 8.7 Hz, 1H), 7.46 (d, J )

15.6 Hz, 1H), 7.63 (dd, J ) 2.4, 8.1 Hz, 1H), 7.79 (dd, J ) 2.4,

8.7 Hz, 1H), 7.84 (d, J ) 8.7 Hz, 1H), 7.96 (d, J ) 2.4 Hz, 1H),

8.18 (t, J ) 6.0 Hz, 1H), 8.46 (d, J ) 2.1 Hz, 1H); MS (DCI/

NH₃) (M + H)⁺at m/z 494, 496. Anal. (C₂₂H₂₁Cl₂N₃O₄S‚

0.77H₂O) C, H, N.

2,4-Dich lor op h en yl 2-((E)-((1-Mor p h olin o)ca r bon yl)-

eth en yl)-4-((E)-((3-(1-p yr r olid in -2-on yl)p r op yla m in oca r -

1

bon yl)eth en yl)p h en yl Su lfid e (25a ): light brown solid; H

NMR (DMSO-d₆, 300 MHz) δ 1.64 (quintet, J ) 6.9 Hz, 2H),

1.92 (quintet, J ) 7.5 Hz, 2H), 2.22 (t, J ) 8.1 Hz, 2H), 3.11-

3.25 (m, 4H), 3.43-3.57 (m, 2H), 3.60 (br m, 6H), 3.72 (br m,

2H), 6.75 (d, J ) 15.6 Hz, 1H), 6.82 (d, J ) 8.9 Hz, 1H), 7.36

(dd, J ) 2.7, 8.9 Hz, 1H), 7.37 (d, J ) 15.3 Hz, 1H), 7.46 (d, J

) 8.9 Hz, 1H), 7.51 (d, J ) 15.6 Hz, 1H), 7.66 (dd, J ) 1.8, 8.9

Hz, 1H), 7.77 (d, J ) 2.7 Hz, 1H), 7.85 (d, J ) 15.6 Hz, 1H),

8.19 (t, J ) 6.0 Hz, 1H), 8.29 (d, J ) 1.8 Hz, 1H); MS (DCI/

NH₃) (M + H)⁺at m/z 587, 589, 591. Anal. (C₂₉H₃₁Cl₂N₃O₄S‚

0.08H₂O) C, H, N.

2,3-Dich lor op h en yl 2-Nitr o-4-((E)-((4-a cetylp ip er a zin -

1-yl)ca r bon yl)eth en yl)p h en yl Su lfid e (28c): light yellow

1

powder; H NMR (DMSO-d₆, 300 MHz) δ 2.04 (s, 3H), 3.42-

3.50 (br m, 4H), 3.50-3.64 (br m, 2H), 3.64-3.79 (br m, 2H),

6.88 (d, J ) 8.7 Hz, 1H), 7.45 (d, J ) 15.6 Hz, 1H), 7.55 (t, J

) 7.65 Hz, 1H), 7.57 (d, J ) 15.6 Hz, 1H), 7.78 (dd, J ) 1.8,

8.1 Hz, 1H), 7.87 (dd, J ) 1.8, 8.1 Hz, 1H), 7.95 (dd, J ) 2.7,

9.0 Hz, 1H), 8.69 (d, J ) 1.8 Hz, 1H); MS (DCI/NH₃) (M +

NH₄)⁺at m/z 497, 499, 501. Anal. (C₂₁H₁₉Cl₂N₃O₄S‚0.85H₂O)

C, H, N.

2,4-Dich lor op h en yl 2-((E)-((1-Mor p h olin o)ca r bon yl)-

eth en yl)-4-((E)-((4-acetylpiper azin -1-yl)car bon yl)eth en yl)-

1

p h en yl Su lfid e (25b): white solid; H NMR (DMSO-d₆, 300

MHz) δ 2.05 (s, 3H), 3.33-3.82 (m, 16H), 6.72 (d, J ) 8.4 Hz,

1H), 7.35 (dd, J ) 2.7, 8.4 Hz, 1H), 7.36 (d, J ) 15.1 Hz, 1H),

7.39 (d, J ) 4.8 Hz, 1H), 7.47 (d, J ) 15.1 Hz, 1H), 7.49 (d, J

) 8.4 Hz, 1H), 7.62 (d, J ) 15.1 Hz, 1H), 7.76 (d, J ) 2.4 Hz,

1H), 7.84 (d, J ) 15.1 Hz, 1H), 7.90 (dd, J ) 1.8, 8.4 Hz, 1H),

8.37 (s, 1H); MS (DCI/NH₃) (M + H)⁺at m/z 574, 576, 578.

Anal. (C₂₈H₂₉Cl₂N₃O₄S‚0.05H₂O) C, H, N.

2,4-Dich lor op h en yl 2-((E)-((1-Mor p h olin o)ca r b on yl)-

et h en yl)-4-((E)-((1-m or p h olin o)ca r b on yl)et h en yl)p h en -

yl Su lfid e (25c): white solid; ¹H NMR (DMSO-d₆, 300 MHz)

δ 3.39-3.83 (m, 16H), 6.77 (d, J ) 8.7 Hz, 1H), 7.35 (dd, J )

2.4, 8.7 Hz, 1H), 7.35 (d, J ) 15.6 Hz, 1H), 7.46 (d, J ) 15.6

Hz, 1H), 7.48 (d, J ) 8.7 Hz, 1H), 7.61 (d, J ) 15.6 Hz, 1H),

7.76 (d, J ) 2.4 Hz, 1H), 7.83 (d, J ) 15.6 Hz, 1H), 7.89 (dd,

J ) 1.8, 8.7 Hz, 1H), 8.36 (d, J ) 1.8 Hz, 1H); MS (DCI/NH₃)

(M + NH₄)⁺at m/z 550, 552, 554. Anal. (C₂₆H₂₆Cl₂N₂O₄S‚

0.17H₂O) C, H, N.

1-Ch lor o-2-n itr o-4-((E)-((4-acetylpiper azin -1-yl)car bon -

yl)eth en yl)ben zen e (27). To a stirred solution of trans-4-

chloro-3-nitrocinnamic acid (26) (1.50 g, 6.59 mmol) and

1-acetylpiperazine (0.89 g, 6.94 mmol) in 20 mL of DMF at

room temperature was added EDAC (1.4 g, 7.30 mmol). The

mixture was then stirred at room temperature for 2 h. TLC

indicated the complete consumption of the acid. Water was

then added to quench the reaction and to precipitate out the

product. The light yellow cinnamide 27 was then collected

through filtration, washed with cold water, dried in a vacuum

oven overnight at 40 °C to give 2.04 g (6.03 mmol, 91.6%) of

the amide as a light yellow solid: ¹H NMR (DMSO-d₆, 300

MHz) δ 2.17 (s, 3H), 3.52-3.60 (br m, 4H), 3.60-3.72 (br m,

2H), 3.72-3.86 (br m, 2H), 6.77 (d, J ) 15.3 Hz, 1H), 7.47 (d,

J ) 15.3 Hz, 1H), 7.80 (d, J ) 8.4 Hz, 1H), 7.90 (d, J ) 2.4,

8.4 Hz, 1H), 8.29 (d, J ) 2.4 Hz, 1H); MS (DCI/NH₃) (M +

NH₄)⁺at m/z 355, 357.

2,4-Dich lor op h en yl 2-Nitr o-4-((E)-((4-a cetylp ip er a zin -

1-yl)ca r bon yl)eth en yl)p h en yl Su lfid e (28b). To a stirred

solution of 4-chloro-3-nitrocinnamide (27) (275 mg, 0.814

mmol) in 1.0 mL of DMF was added potassium carbonate (169

mg, 1.22 mmol), followed by the dropwise addition of 2,4-

dichlorothiophenol (146 mg, 0.815 mmol). The mixture was

then stirred at room temperature for 60 min. Completion of

the reaction was indicated by the TLC, and water was then

added to precipitate the product. Filtration, washing with cold

water, and drying in a vacuum oven afforded 350 mg (0.728

mmol, 89%) of compound 28b as a light yellow solid: ¹H NMR

(DMSO-d₆, 300 MHz) δ 2.05 (s, 3H), 3.42-3.50 (br m, 4H),

3.50-3.64 (br m, 2H), 3.64-3.79 (br m, 2H), 6.83 (d, J ) 8.7

2-Meth ylph en yl 2-Nitr o-4-((E)-((4-acetylpiper azin -1-yl)-

ca r bon yl)eth en yl)p h en yl Su lfid e (28d ): light yellow pow-

der; ¹H NMR (DMSO-d₆, 300 MHz) δ 2.03 (s, 3H), 2.29 (s, 3H),

3.47 (br m, 4H), 3.53 (br m, 1H), 3.60 (br m, 1H), 3.67 (br m,

1H), 3.83 (br m, 1H), 6.64 (d, J ) 8.7 Hz, 1H), 7.40 (d, J )

15.0 Hz, 1H), 7.36-7.42 (m, 1H), 7.46-7.57 (m, 3H), 7.63 (d,

J ) 6.9 Hz, 1H), 7.89 (dd, J ) 2.4, 9.0 Hz, 1H), 8.66 (d, J )

2.4 Hz, 1H); MS (APCI) (M + H)⁺at m/z 426. Anal.

(C₂₂H₂₃N₃O₄S‚0.1H₂O) C, H, N.

2-Ch lor oph en yl 2-Nitr o-4-((E)-((4-acetylpiper azin -1-yl)-

ca r bon yl)eth en yl)p h en yl Su lfid e (28e): light yellow pow-

der; ¹H NMR (DMSO-d₆, 300 MHz) δ 2.04 (s, 3H), 3.47 (br m,

4H), 3.52 (br m, 1H), 3.60 (br m, 1H), 3.68 (br m, 1H), 3.73 (br

m, 1H), 6.75 (d, J ) 9.0 Hz, 1H), 7.43 (d, J ) 15.3 Hz, 1H),

7.54 (d, J ) 15.3 Hz, 1H), 7.55 (dd, J ) 1.8, 8.1 Hz, 1H), 7.64

(t, J ) 1.8, 8.1 Hz, 1H), 7.76 (d, J ) 1.8, 8.1 Hz, 1H), 7.82 (d,

J ) 1.8, 8.1 Hz, 1H), 7.93 (dd, J ) 2.4, 9.0 Hz, 1H), 8.68 (d, J

) 2.4 Hz, 1H); MS (APCI) (M + H)⁺at m/z 446, 448, 450. Anal.

(C₂₁H₂₀ClN₃O₄S‚0.03H₂O) C, H, N.

2-Am in op h en yl 2-Nitr o-4-((E)-((4-a cetylp ip er a zin -1-yl-

)ca r bon yl)eth en yl)p h en yl Su lfid e (28f): light yellow pow-

der; ¹H NMR (DMSO-d₆, 300 MHz) δ 2.04 (s, 3H), 3.47 (br m,

4H), 3.52 (br m, 1H), 3.60 (br m, 1H), 3.68 (br m, 1H), 3.74 (br

m, 1H), 5.58 (s, 2H), 6.65 (td, J ) 1.5, 15.0 Hz, 1H), 6.72 (dd,

J ) 1.5, 8.7 Hz, 1H), 7.00 (dd, J ) 1.8, 8.7 Hz, 1H), 7.27 (t, J

) 1.5, 8.6 Hz, 1H), 7.36 (dd, J ) 1.5, 8.7 Hz, 1H), 7.39 (d, J )

15.3 Hz, 1H), 7.53 (d, J ) 15.3 Hz, 1H), 7.89 (dd, J ) 1.8, 8.7

Hz, 1H), 8.64 (d, J ) 1.8 Hz, 1H); MS (APCI) (M + H)⁺at m/z

427. Anal. (C₂₁H₂₂N₄O₄S‚0.43H₂O) C, H, N.

2-Eth ylp h en yl 2-Nitr o-4-((E)-((4-a cetylp ip er a zin -1-yl)-

ca r bon yl)eth en yl)p h en yl Su lfid e (28g): light yellow pow-

der; ¹H NMR (DMSO-d₆, 300 MHz) δ 1.01 (t, J ) 7.65 Hz, 3H),

2.04 (s, 3H), 2.69 (q, J ) 7.65 Hz, 2H), 3.47 (br m, 4H), 3.52

(br m, 1H), 3.59 (br m, 1H), 3.67 (br m, 1H), 3.73 (br m, 1H),

6.64 (d, J ) 8.7 Hz, 1H), 7.38 (dd, J ) 2.4, 7.5 Hz, 1H), 7.40

(d, J ) 15.6 Hz, 1H), 7.50-7.61 (m, 3H), 7.53 (d, J ) 15.6 Hz,

1H), 7.89 (dd, J ) 2.4, 8.7 Hz, 1H), 8.64 (d, J ) 2.4 Hz, 1H);

MS (APCI) (M + Cl)^-at m/z 474, 476. Anal. (C₂₃H₂₅N₃O₄S‚

0.59H₂O) C, H, N.

2-Isop r op ylp h en yl 2-Nitr o-4-((E)-((4-a cetylp ip er a zin -

1-yl)ca r bon yl)eth en yl)p h en yl Su lfid e (28h ): light yellow

powder; ¹H NMR (DMSO-d₆, 300 MHz) δ 1.05 (d, J ) 6.9 Hz,

6H), 2.04 (s, 3H), 3.47 (br m, 4H), 3.52 (br m, 1H), 3.60 (br m,

1H), 3.67 (br m, 1H), 3.72 (br m, 1H), 6.64 (d, J ) 8.4 Hz, 1H),

7.34-7.41 (m, 2H), 7.39 (d, J ) 15.3 Hz, 1H), 7.52 (d, J ) 15.3

Hz, 1H), 7.56-7.73 (m, 2H), 7.90 (dd, J ) 2.1, 8.7 Hz, 1H),

Novel p-Arylthio Cinnamides

J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 21 4039

8.64 (d, J ) 2.1 Hz, 1H); MS (APCI) (M + NH₄)⁺at m/z 471.

Anal. (C₂₄H₂₇N₃O₄S‚0.21H₂O) C, H, N.

) 2.1 Hz, 1H), 8.13 (t, J ) 6.0 Hz, 1H); MS (ESI) (M + H)⁺at

m/z 527, 529, 531, 533. Anal. (C₂₂H₂₁BrCl₂N₂O₂S‚0.18H₂O) C,

H, N.

2-ter t-Bu tylp h en yl 2-Nitr o-4-((E)-((4-a cetylp ip er a zin -

1-yl)ca r bon yl)eth en yl)p h en yl Su lfid e (28i): light yellow

powder; ¹H NMR (DMSO-d₆, 300 MHz) δ 1.46 (s, 9H), 2.04 (s,

3H), 3.47 (br m, 4H), 3.52 (br m, 1H), 3.60 (br m, 1H), 3.67 (br

m, 1H), 3.73 (br m, 1H), 6.68 (d, J ) 8.7 Hz, 1H), 7.35 (t, J )

7.5 Hz, 1H), 7.39 (d, J ) 15.3 Hz, 1H), 7.45-7.57 (m, 2H), 7.50

(d, J ) 15.3 Hz, 1H), 7.65 (d, J ) 8.1 Hz, 1H), 7.88 (dd, J )

2.4, 8.7 Hz, 1H), 8.64 (d, J ) 2.4 Hz, 1H); MS (APCI) (M +

NH₄)⁺at m/z 485. Anal. (C₂₅H₂₉N₃O₄S‚0.07H₂O) C, H, N.

2-F or m ylp h en yl 2-Nitr o-4-((E)-((4-a cetylp ip er a zin -1-

yl)ca r bon yl)eth en yl)p h en yl Su lfid e (28j): yellow solid; ¹H

NMR (DMSO-d₆, 300 MHz) δ 2.04 (s, 3H), 3.47 (br m, 4H),

3.52 (br m, 1H), 3.60 (br m, 1H), 3.68 (br m, 1H), 3.74 (br m,

1H), 6.85 (d, J ) 8.4 Hz, 1H), 7.44 (d, J ) 15.6 Hz, 1H), 7.55

(d, J ) 15.6 Hz, 1H), 7.61 (d, J ) 7.5 Hz, 1H), 7.73 (t, J ) 7.5

Hz, 1H), 7.80 (td, J ) 2.4, 7.5 Hz, 1H), 7.92 (dd, J ) 2.1, 9.0

Hz, 1H), 8.04 (dd, J ) 2.4, 7.5 Hz, 1H), 8.66 (d, J ) 2.1 Hz,

1H), 10.29 (s, 1H); MS (APCI) (M + Cl)^-at m/z 474, 476. Anal.

(C₂₂H₂₁N₃O₅S‚0.33H₂O) C, H, N.

2-Dim eth yla m in op h en yl 2-Nitr o-4-((E)-((4-a cetylp ip -

er a zin -1-yl)ca r bon yl)eth en yl)p h en yl Su lfid e (28k ). To a

stirred solution of aniline 28f (21 mg, 0.049 mmol) in 1 mL of

ethanol was added Me₂SO₄(14.0 µL, 0.15 mmol) followed by

saturated Na₂CO₃(25 mL). The mixture was then refluxed for

1 day. The reaction mixture was allowed to cool to ambient

temperature, partitioned between EtOAc and water. The

organic layer was washed with brine, dried over Na₂SO₄,

filtered, concentrated under reduced pressure. The residue was

2,4-Dich lor oph en yl 2-((3-(1-Mor ph olin o)pr opyl)am in o)-

4-((E)-((3-(1-p yr r olid in -2-on yl)p r op yla m in o)ca r b on yl)-

et h en yl)p h en yl Su lfid e (31): light brown solid; ¹H NMR

(CDCl₃, 300 MHz) δ 1.70-1.89 (m, 4H), 2.06 (quintet, J ) 7.65

Hz, 4H), 2.24-2.50 (m, 8H), 3.23 (p, J ) 6.0 Hz, 2H), 3.29-

3.48 (m, 4H), 3.59-3.83 (m, 4H), 6.56 (d, J ) 8.4 Hz, 1H),

6.77-6.90 (m, 2H), 7.02 (dd, J ) 2.8, 8.4 Hz, 1H), 7.09-7.18

(m, 1H), 7.33-7.39 (m, 2H), 7.42 (d, J ) 8.4 Hz, 1H), 7.57 (d,

J ) 15.6 Hz, 1H); MS (ESI) (M + H)⁺at m/z 591, 593, 595.

Anal. (C₂₉H₃₆Cl₂N₄O₃S‚1.02TFA) C, H, N.

5-Br om o-2-[(2,4-dich lor oph en yl)th io]ben zaldeh yde (32).

Prepared analogously to compound 5, substituting 2-chloroni-

trobenzene with 5-bromo-2-fluorobenzaldehyde: white solid;

¹H NMR (DMSO-d₆, 300 MHz) δ 7.21 (d, J ) 2.7 Hz, 1H), 7.25

(d, J ) 2.7 Hz, 1H), 7.40 (dd, J ) 2.7, 8.4 Hz, 1H), 7.76 (dd, J

) 2.1, 8.4 Hz, 1H), 8.39 (d, J ) 2.1 Hz, 1H), 9.99 (s, 1H); MS

(DCI/NH₃) (M + H)⁺at m/z 360, 362, 364.

2,4-Dich lor op h en yl 2-F or m yl-4-((E)-((1-m or p h olin o)-

ca r bon yl)eth en yl)p h en yl Su lfid e (33): white solid; ¹H NMR

(DMSO-d₆, 300 MHz) δ 3.60 (br m, 6H), 3.71 (br m, 2H), 6.82

(d, J ) 8.7 Hz, 1H), 7.35 (d, J ) 15.6 Hz, 1H), 7.54 (d, J )

15.6 Hz, 1H), 7.55 (dd, J ) 2.4, 8.7 Hz, 1H), 7.61 (d, J ) 8.7

Hz, 1H), 7.86 (dd, J ) 2.4, 8.4 Hz, 1H), 7.91 (d, J ) 2.4 Hz,

1H), 8.41 (d, J ) 2.1 Hz, 1H), 10.19 (s, 1H); MS (DCI/NH₃) (M

+ H)⁺at m/z 422, 424, 426. Anal. (C₂₀H₁₇Cl₂NO₃S) C, H, N.

2,4-Dich lor op h en yl 2-Ca r boxy-4-((E)-((1-m or p h olin o)-

ca r bon yl)eth en yl)p h en yl Su lfid e (34). To a stirred suspen-

sion of the Ag₂O (110 mg, 0.47 mmol) in 2 mL of H₂O at room

temperature was added the aldehyde 33 (100 mg, 0.24 mmol),

followed by 3 N NaOH (0.16 mL, 0.47 mmol). The mixture was

stirred overnight, then filtered through Celite, washed with 3

N NaOH and H₂O. The filtrate was acidified, the precipitates

collected through filtration, dried in a vacuum oven to give 75

mg (0.17 mmol, 71%) of the acid 34 as a white solid: ¹H NMR

(DMSO-d₆, 300 MHz) δ 3.50-3.62 (m, 6H), 3.62-3.73 (m, 2H),

6.50 (d, J ) 8.4 Hz, 1H), 7.16 (d, J ) 15.0 Hz, 1H), 7.44 (d, J

) 15.0 Hz, 1H), 7.51 (d, J ) 2.8, 8.4 Hz, 1H), 7.64 (d, J ) 8.4

Hz, 1H), 7.82 (d, J ) 2.8 Hz, 1H), 8.11 (d, J ) 1.8 Hz, 1H); MS

(DCI) (M + H)⁺at m/z 438, 440, 442. Anal. (C₂₀H₁₇Cl₂NO₄S‚

0.33H₂O) C, H, N.

2,4-Dich lor op h en yl 2-Ca r boxa m id o-4-((E)-((3-(1-m or -

p h olin o)p r op yla m in o)ca r bon yl)eth en yl)p h en yl Su lfid e

(35). A mixture of acid 34 (35 mg, 0.080 mmol), EDAC (21

mg, 0.112 mmol), HOBT (14 mg, 0.096 mmol), NH₄Cl (22 mg,

0.40 mmol), and Et₃N (28.5 µL, 0.20 mmol) in 2.0 mL of DMF

was stirred at room temperature for overnight. Water was

added to the reaction mixture, and the precipitates were

collected through filtration, washed with H₂O, and dried in a

vacuum oven to give 26 mg (0.059 mmol, 74% yield) of amide

35 as a white solid: ¹H NMR (DMSO-d₆, 300 MHz) δ 3.48-

3.66 (m, 6H), 3.66-3.78 (m, 2H), 6.86 (d, J ) 8.4 Hz, 1H), 7.29

(d, J ) 15.6 Hz, 1H), 7.43-7.52 (m, 3H), 7.64 (dd, J ) 2.4, 8.7

Hz, 1H), 7.79 (d, J ) 2.7 Hz, 1H), 7.99 (d, J ) 2.4 Hz, 1H); MS

(ESI) (M + H)⁺at m/z 437, 439, 441. Anal. (C₂₀H₁₈Cl₂N₂O₃S‚

0.72H₂O) C, H, N.

ICAM-1/LF A-1 Bioch em ica l In ter a ction Assa y. In the

biochemical assay, 100 µL of anti-LFA-1 antibody (ICOS Corp.)

at a concentration of 5 µg/mL in Dulbecco’s phosphate-buffered

saline (D-PBS) was used to coat wells of a 96-well microtiter

plate overnight at 4 °C. The wells were then washed twice with

wash buffer (D-PBS without Ca²⁺or Mg²⁺, 0.05% Tween 20)

and blocked by addition of 200 µL of D-PBS, 5% fish skin

gelatin. Recombinant LFA-1 (100 µL of 0.7 µg/mL; ICOS Corp.)

in D-PBS was then added to each well. Incubation continued

for 1 h at room temperature and the wells were washed twice

with wash buffer. Serial dilutions of compounds being assayed

as ICAM-1/LFA-1 antagonists, prepared as 10 mM stock

solutions in dimethyl sulfoxide (DMSO), were diluted in

D-PBS, 2 mM MgCl₂, 1% fish skin gelatin and 50 µL of each

dilution was added to duplicate wells. This was followed by

addition of 50 µL of 0.8 µg/mL biotinylated recombinant ICAM-

then purified on

a preparative HPLC to give 10 mg of

compound 28k (0.022 mmol, 45% yield) as a light yellow

solid: ¹H NMR (CDCl₃, 300 MHz) δ 2.16 (s, 3H), 2.83 (s, 3H),

3.32 (br s, 3H), 3.47-3.85 (m, 8H), 6.75 (d, J ) 8.4 Hz, 1H),

6.78 (d, J ) 8.4 Hz, 1H), 6.82 (d, J ) 8.4 Hz, 1H), 6.89 (d, J )

15.6 Hz, 1H), 7.40-7.51 (m, 3H), 7.64 (d, J ) 15.6 Hz, 1H),

8.45 (d, J ) 1.8 Hz, 1H); MS (APCI) (M + H)⁺at m/z 454.

Anal. (C₂₃H₂₆N₄O₄S‚0.18H₂O) C, H, N.

2-Ca r boxa m id op h en yl 2-Nitr o-4-((E)-((4-a cetylp ip er -

a zin -1-yl)ca r bon yl)et h en yl)p h en yl Su lfid e (28l): yellow

solid; ¹H NMR (DMSO-d₆, 300 MHz) δ 2.04 (s, 3H), 3.43-3.80

(m, 8H), 6.93 (d, J ) 8.7 Hz, 1H), 7.42 (d, J ) 15.6 Hz, 1H),

7.49-7.63 (m, 4H), 7.89 (dd, J ) 2.1, 8.7 Hz, 1H), 7.93 (s, 1H),

8.59 (d, J ) 2.1 Hz, 1H); MS (APCI) (M + Cl)^-at m/z 489,

491. Anal. (C₂₂H₂₂N₄O₅S‚0.33H₂O) C, H, N.

2,4-Dich lor op h en yl 2-Am in o-4-((E)-((3-(1-p yr r olid in -2-

on yl)p r op yla m in o)ca r bon yl)eth en yl)p h en yl Su lfid e (29):

yellow solid; ¹H NMR (DMSO-d₆, 300 MHz) δ 1.65 (quintet, J

) 7.0 Hz, 2H), 1.92 (quintet, J ) 7.5 Hz, 2H), 2.22 (t, J ) 7.5

Hz, 2H), 3.15 (q, J ) 7.0 Hz, 2H), 3.21 (t, J ) 7.5 Hz, 2H),

3.30-3.43 (t, J ) 7.0 Hz, 2H), 5.63 (s, 2H), 6.55 (d, J ) 8.4

Hz, 1H), 6.67 (d, J ) 15.6 Hz, 1H), 6.83 (dd, J ) 1.8, 8.4 Hz,

1H), 6.99 (d, J ) 1.8 Hz, 1H), 7.25-7.38 (m, 3H), 7.66 (d, J )

1.8 Hz, 1H), 8.20 (t, J ) 5.85 Hz, 1H); MS (DCI) (M + H)⁺at

m/z 464, 466, 468. Anal. (C₂₂H₂₃Cl₂N₃O₂S‚0.56TFA) C, H, N.

2,4-Dich lor op h en yl 2-Br om o-4-((E)-((3-(1-p yr r olid in -2-

on yl)p r op yla m in o)ca r bon yl)eth en yl)p h en yl Su lfid e (30).

To a stirred solution of tert-butyl nitrite (57 µL, 0.48 mmol),

CuBr₂(87 mg, 0.39 mmol) in 2.0 mL of CH₃CN at room

temperature was added a solution of aniline 29 (150 mg, 0.323

mmol) in 1.0 mL of CH₃CN. The dark green solution was then

heated at 65 °C under nitrogen atmosphere for 90 min. The

reaction mixture was then allowed to cool to room temperature,

partitioned between EtOAc and 3 N HCl. The organic layer

was then washed with brine, dried over Na₂SO₄, condensed

in vacuuo. The crude product was then purified with a

preparative HPLC to give 50 mg (0.094 mmol, 29%) of bromide

30 as a light brown solid: ¹H NMR (DMSO-d₆, 300 MHz) δ

1.63 (quintet, J ) 7.2 Hz, 2H), 1.91 (quintet, J ) 8.4 Hz, 2H),

2.22 (t, J ) 8.4 Hz, 2H), 3.09-3.47 (m, 6H), 6.67 (d, J ) 15.3

Hz, 1H), 7.07 (d, J ) 8.4 Hz, 1H), 7.32 (d, J ) 8.7 Hz, 1H),

7.38 (d, J ) 15.3 Hz, 1H), 7.50 (dd, J ) 2.4, 8.7 Hz, 1H), 7.57

(dd, J ) 2.1, 8.4 Hz, 1H), 7.86 (d, J ) 2.4 Hz, 1H), 7.96 (d, J

4040 J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 21

Liu et al.

1/Ig (ICOS Corp.) to the wells and the plates were incubated

at room temperature for 1 h. The wells were then washed twice

with wash buffer and 100 µL of europium-labeled streptavidin

(Wallac Oy) diluted 1:100 in Delfia assay buffer (Wallac Oy)

was added to the wells. Incubation proceeded for 1 h at room

temperature. The wells were washed eight times with wash

buffer and 100 µL of enhancement solution (Wallac Oy, cat.

No. 1244-105) was added to each well. Incubation proceeded

for 5 min with constant mixing. Time-resolved fluorimetry

measurements were made using the Victor 1420 multilabel

counter (Wallac Oy) and the percent inhibition of each

candidate compound was calculated using the following equa-

tion: % inhibition ) 100 × {1 - (average OD with compound

minus background)/(average OD without compound minus

background)}, where “background” refers to wells that are not

coated with anti-LFA-1 antibody.

Ack n ow led gm en t. The authors thank the Abbott

Analytical Department for assistance in acquiring H

NMR and mass spectra.

1

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J M0002782

Article Doi

Discovery of novel p-arylthio cinnamides as antagonists of leukocyte function-associated antigen-1/intracellular adhesion molecule-1 interaction. 1. Identification of an additional binding pocket based on an anilino diaryl sulfide lead

DOI: 10.1021/jm0002782

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Full text of DOI:10.1021/jm0002782

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