JOURNAL OF OCULAR PHARMACOLOGY AND THERAPEUTICS

Volume 22, Number 6, 2006

© Mary Ann Liebert, Inc.

Retinal Penetration of Calpain Inhibitors in Rats

After Oral Administration

YOSHIHISA SHIRASAKI,1 MASAZUMI YAMAGUCHI,1 and HIROYUKI MIYASHITA2

ABSTRACT

Calpain-mediated proteolysis has been involved in neuronal cell death of retinal neuro-
logical degeneration. An aldehyde-based calpain inhibitor, SJA6017 (1), was effective fol-
lowing oral administration in a rat retinal ischemia model but had low oral bioavailability.
The aim of this study was to identify calpain inhibitors with good retinal penetration after
oral dosing. The orally bioavailable inhibitors, hemiacetal 3 (SNJ-1715), amphipathic ke-
toamide 5 (SNJ-1945), and pyridine ketoamide 6 (SNJ-2008), were evaluated for their retinal
pharmacokinetic (PK) profiles. The retinal drug exposure of these inhibitors was more than
tenfold higher than 1. Among these compounds, 5 exhibited the most favorable retinal PK
properties, such as good penetration and long half-life. Comparisons of 5 and the structurally
related ketoamide 6 suggested that the presence of a methoxy diethylene glycol moiety re-
sulted in the inhibitor with high penetration into the retina and the sustained high retinal
levels. Ketoamide 5 was selected as the development candidate for the treatment of retinal
diseases.

INTRODUCTION mice, which are an accepted animal model for

C ALPAINS ARE CALCIUM-ACTIVATED intracellular
cysteine endoproteases, and 15 genes of
isozymes have, so far, been identified.1–3 Two ma-
jor isoforms, -calpain (calpain I) and m-calpain
(calpain II), are ubiquitously present in mam-
malian cells. These enzymes have been impli-
cated in numerous neurological disorders, in-
cluding stroke, Alzheimer’s disease, central
nervous system diseases, multiple sclerosis,
spinal cord injury, and traumatic brain injury
(TBI).4–7 Previously, our researchers have re-
ported that an activation of calpains occurred in
retinal neuronal cell death during retinal is-
chemia-reperfusion8 and acute ocular hyperten-
sion in rats.9 It has also been shown that calpains
are involved in photoreceptor cell death in rd1
photoreceptor degeneration in retinitis pigmen-
tosa.10,11 These results suggest that calpains are a
potential target for retinal neurological disorders,
such as glaucoma and retinitis pigmentosa.
A number of calpain inhibitors have been
synthesized and some of them have demon-
strated neuroprotective efficacy in several animal
pharmacological models through injections.12–19
Because most of retinal diseases are chronic dis-
eases, intravenous (i.v.) administration is incon-
venient. Topical instillation to the eyes would be
desirable, but it cannot readily deliver drugs into
the retina because of several pathological and
anatomical barriers, such as nasolacrimal drain-
age, metabolism, protein binding, and lens bar-
rier.20 Therefore, oral drugs are preferable for the
treatment of the retinal diseases. Our group has

1Senju Pharmaceutical Co., Ltd., Hyogo, Japan.

2Faculty of Pharmaceutical Sciences, Kumamoto University, Kumamoto, Japan.

417
418
reported that oral administration of a highly po-
tent inhibitor, dipeptidyl aldehyde 1 (SJA6017),21,22
showed neuroprotective efficacy in the rat retinal
ischemia model.3 However, a relatively high dose
(500 mg/kg) was required, presumably owing to
low oral bioavailability (BA), which was ascribed
to a metabolically labile aldehyde moiety and low
water solubility.23 Consequently, we investigated
whether replacing the aldehyde warhead with
more chemically stable warheads, such as -ke-
toamide and cyclic hemiacetal moiety, could im-
prove the oral BA (Fig. 1). The cyclic hemiacetal
derivative 2 showed a high metabolic stability
with adequate aqueous solubility. Further opti-
mization of 2 for inhibitory activity provided an
oral available inhibitor 3 (SNJ-1715) with an ac-
ceptable potency.23,24 On the other hand, -ke-
toamide analog 425 (Fig. 1) was equipotent to the
original aldehyde 1, and more membrane-per-
meable and metabolically stable than 1, but it
showed poor oral absorption in monkeys, likely
owing to its extremely low aqueous solubility.
FIG. 1. Drug design concept of novel calpain inhibitors.
SHIRASAKI ET AL.
We have improved the aqueous solubility of 4 by
incorporating a nonionic amphiphile and identi-
fied a dipeptidyl -ketoamide 5 (SNJ-1945) that
showed good plasma exposure in monkeys (Fig.
1).25 In addition, introduction of a pyridine moi-
ety as a basic water-soluble moiety also provided
a potent and orally bioavailable ketoamide 6
(SNJ-2008) (Fig. 1).26
Our aim was to discover orally available cal-
pain inhibitors with good retinal penetration.
Drug penetration into the retina is restricted by
the inner and outer blood-retinal barriers (BRB).
Therefore, the plasma drug levels are not always
parallel to the retinal levels.27,28 Although the in-
hibitors 3, 5, and 6 showed good plasma expo-
sure following oral administration, it is uncertain
whether the inhibitors penetrate into the retina.
In this study, we evaluated the inhibitory activi-
ties, oral BA, and retinal penetration in rats of cal-
pain inhibitors, original aldehyde 1, hemiacetal 3,
amphipathic ketoamide 5, and pyridine ke-
toamide 6.
RETINAL PENETRATION OF CALPAIN INHIBITORS
METHODS
Materials
The calpain inhibitors, SJA6017 (1),21,22
SNJ-1715 (3),23,24 SNJ-1945 (5),25 and SNJ-2008
(6),26 were synthesized, as reported previously.
Sodium carboxymethyl cellulose (CMC-Na) and
polyethylene glycol 300 were purchased from
Wako Pure Chemical Industries, Ltd. (Osaka,
Japan).
Calpain inhibition assay
Calpain inhibition assays were performed, as
described in the previous paper,29 using com-
mercial -calpain (human erythrocyte; Cal-
biochem, San Diego, CA) and m-calpain (porcine
kidney; Calbiochem). Assay solution, including
80 mM Tris-HCl (pH 7.4), 15 mM -mercap-
toethanol, and 15 nmol of enzyme was used. The
assay solution (150 L), 0.5 mM Boc-Val-Leu-Lys-
AMC (50 L; Bachem, Budendorf, Switzerland),
and dimethyl sulfoxide (2.5 L) including in-
hibitor were placed in each well of 96-well plates.
Reaction was started by the addition of 25 mM
CaCl2 (50 L) to a test well and 1 mM of ethyl-
enediaminetetraacetic acid (50 L) in a blank
well. Enzyme activity was determined by the in-
crease of fluorescence (ex  360 nm, em  440
nm) monitored at 37°C, using a CYTO FLUOR
multiwell plate reader (Perspective Biosystems,
Foster City, CA). Percent enzyme inhibition was
determined by a comparison of this activity to
that of a solution without inhibitor.
Dose preparation
For pharmacokinetic studies, the i.v. dose of 1,
3, 5, or 6 (3 mg/kg) was administered in a vehi-
cle consisting of polyethylene glycol 300/saline
(3:7) at a concentration of 0.6 mg/mL (5 mL/kg).
The oral dose (10 mg/kg for 3, 5, and 6) was ad-
ministered in a suspension in 0.5% CMC-Na so-
lution at a concentration of 2 mg/mL (5 mL/kg).
Compound 1 for oral dose (500 mg/kg) was for-
mulated as a suspension in 0.5% CMC-Na solu-
tion at a concentration of 100 mg/mL (5 mL/kg).
Pharmacokinetic and retinal penetration studies
Male Sprague-Dawley rats (weight 200–250 g)
were purchased from Charles River Laboratories
Japan, Inc. (Yokohama, Japan). All aspects of han-
419
dling, housing, and experimentation conformed to
the ARVO Resolution for the Use of Animals in
Ophthalmic and Vision Research. Animals were
fasted overnight before dosing in the case of oral
dosing. Animals were allowed unrestricted access
to water. Rats (n  4–5 per time point) were dosed
with compounds 1, 3, 5, and 6 orally by gavage at
a dose of 10 or 500 mg/kg. They were also dosed
with 1, 3, 5, and 6 at 3 mg/kg by bolus injection
through a tail vein. The animals were anesthetized
with isoflurane at a predetermined time (0.1 [i.v.
only], 0.25, 0.5, 1, 2, 4, and 8 h after dosing), and
blood samples (5–6 mL) were collected from the
abdominal aorta into heparinized syringes. Then,
these animals, including the eyes, were perfused
with saline (12 mL). The eye globes were enucle-
ated, and the retinas were excised. Plasma was pre-
pared by centrifugation of the blood. All plasma
and retina samples were frozen and stored at
30°C until analysis.
Preparation of plasma samples
Plasma samples (0.2 mL) were mixed with
phosphate-buffered saline (1 mL) and the solu-
tion was transferred to OASIS® HLB (60 mg) car-
tridges (Waters; Tokyo, Japan). The solid phase
was washed with 5% aqueous MeOH (1 mL) and
eluted MeOH (1 mL). The eluent was evaporated
to dryness in vacuo at 40°C, reconstituted with 1
mL of mobile phase (MeOH/H2O/HCO2H 
40:60:0.1 for 1, 5, and 6 or MeCN/H2O  70:30
for 3) and 10-L aliquots of these solutions were
analyzed by liquid chromatography/mass spec-
trometry/mass spectrometry (LC-MS/MS; de-
scribed below).
Preparation of retinal samples
The retinas were weighted and homogenized
in MeOH (5 mL). The homogenates were cen-
trifuged at 3000g for 10 min. The supernatant (4
mL) was transferred to glass test tubes and evap-
orated to dryness in vacuo at 40°C. The residue
was reconstituted with mobile phase (1 mL),
transferred to 1.5-mL tubes, and centrifuged at
10,000g for 10 min. Then, 10-L aliquots of the
supernatants were analyzed by LC-MS/MS.
LC-MS/MS analysis of plasma and
retina samples
The plasma and retinal concentration of each
test compound were determined by turbo ion
420 SHIRASAKI ET AL.

spray on an API 4000 triple-quadrupole mass Determination of pharmacokinetic parameters

spectrometer (Applied Biosystems/MDS-Sciex;
The pharmacokinetic parameters were deter-
Ontario, Canada) equipped with a turbo ion
mined from the resulting mean plasma or retinal
spray source using multiple reaction monitoring
concentration versus time profiles by using the
(MRM). Chromatography was performed on a
noncompartmental model analysis (WinNonlin
NANOSPACE SI-2 HPLC system (Shiseido;
version 2.1; Pharsight, Mountain View, CA). The
Tokyo, Japan) with Shiseido Capcell pak® C18
calculated parameters are maximal concentration
MG-II column (75  1.5 mm, 5 m) at 40°C, us-
of compounds (Cmax), time required to reach Cmax
ing a mobile phase (MeOH/H2O/HCO2H 
(Tmax), plasma and retina terminal half-lives (T1/2
40:60:0.1) for compounds 1, 5, and 6 or Shiseido
and t1/2, respectively), the area under the curves
Capcell pak C18 ACR (150  1.5 mm, 3 m) at
from time 0 to infinity (AUC) in plasma or retina,
40°C, using a mobile phase (MeCN/H2O  70:30)
the volume of distribution at steady state (Vss),
for 3 at a flow rate of 0.20 mL/min. The dwell time
and plasma total clearance (Cl). Oral bioavail-
for the MRM was 150 ms. A peak was defined by
ability (F) was calculated from the plasma AUC
manual integration. The negative ion mode was
values after oral and i.v. dosing, corrected by the
used for 1 and 3. The compounds 5 and 6 were
ratio of the doses administrated.
measured by positive ion mode. Multiple reaction
monitoring using the product ions (m/z 371.1 
158.5 for 1, m/z 350.2  117.0 for 3, m/z 492.2 
102.8 for 5, and m/z 495.1  106.1 for 6) was used
RESULTS
for quantification. The levels of these compounds
in plasma or retina samples were determined by
Inhibitory activities
using standard curves in which control plasma or
control retina was spiked with increasing concen- Previously, we evaluated the calpain in-
trations of these compounds (5–50 ng for plasma, hibitory activities of some inhibitors in a dye
2.5–50 ng for retina). A regression fit, 1/X, was used assay method.22,24,25 In this study, the activi-
to quantify the unknowns. The limits of quantifi- ties of aldehyde 1, hemiacetal 3, amphipathic
cation for these compounds were 5 ng/mL in ketoamide 5, and pyridine ketoamide 6 were
plasma and 2.5 ng/g of tissue in retina, respec- determined under the same conditions, using
tively. Data reduction were performed using Sciex a fluorogenic substrate to compare the poten-
Analyst versus 1.3 software (Applied Biosystems/ tial retinal efficacy. The data are summarized
MDS-Sciex, Ontario, Canada), and the mean and in Table 1. The inhibitory activities of 3, 5, and
standard deviation was calculated by using Micro- 6 are acceptable and comparable to original
soft Excel® software (Microsoft, Tokyo, Japan). aldehyde 1.

TABLE 1. ENZYME INHIBITORY ACTIVITY, PHYSICOCHEMICAL PROPERTIES OF CALPAIN INHIBITORS

IC50 (M)a
Metab.b
Compound -calpain m-calpain (%) Mw c cLogD7d

1 0.028 0.023 3e 372 3.21

3 0.087 0.071 100e 351 0.51
5 0.062 0.045 41f 491 2.27
6 0.029 0.017 23g 494 3.31
aTheIC50 values were determined by the fluorescence assay method (n  2).
bMetabolic stability represented in residual percent after incubation with human
S9 for 0.5 h at 37°C.
cMolecular
weight.
dThe dissociative partition coefficient ACD cLogD was calculated using Advanced Chemistry Development Soft-
7
ware V4.76 for Solaris (ACD/Labs).
eReference 23.
fReference 25.
gReference 26.
RETINAL PENETRATION OF CALPAIN INHIBITORS 421

TABLE 2. PLASMA PK PARAMETERS OF CALPAIN INHIBITORS IN RATS

Parameters 1 3 5 6

Route i.v. p.o. i.v. p.o. i.v. p.o. i.v. p.o.

Dose (mg/kg) 3.0 500/(10)g 30.0 100.0 30.0 100.0 30.0 100.0
The number of animals 4.0 4 50.0 50.0 50.0 50.0 50.0 50.0
(n/time point)
AUCa (M  h) 1.50 45/(0.90)g 2.60 5.70 2.30 7.40 4.50 6.80
g  h/mL 0.56 16/(0.34)g 7.40 2.00 1.10 3.60 2.20 3.40
Cmax (M) — 9.4/(0.19)g — 5.60 — 3.50 — 8.60
(g/mL) 3.5/(0.70)g 2.00 1.70 4.30
Tmax (h) — 1.0 — 0.25 — 0.25 — 0.25
T1/2b (h) 0.70 — 1.90 — 3.80 — 2.10 —
zc (h1) 0.99 — 0.39 0.18 0.33
Vssd (L/kg) 6.10 — 2.30 — 4.30 — 0.98 —
Cle (mL/min/kg) 890.0 — 550.0 — 440.0 — 220.0 —
Ff (%) 18 66 96 45

PK, pharmacokinetic; AUC, area under the curves; i.v., intravenous; p.o., per os.
aThe area under the curves for 0 h to infinity.
bThe terminal half-life.
cThe terminal phase elimination rate constant.
dThe steady-state volume of distribution.
eThe total clearance.
fOral bioavailability.
gThe values dose-normalized to 10 mg/kg.

Oral plasma bioavailability P3 moiety, ketoamide 6 demonstrated the signif-

icantly lower Vss value (0.98 L/kg) than 5 (Vss 
The oral plasma bioavailability (BA) of 1, 3, 5,
4.3 L/kg).
and 6 were investigated in Sprague-Dawley rats
dosed i.v. at 3 mg/kg8 and orally at 10 mg/kg
for 3, 5, and 6 or 500 mg/kg23,26 for 1 (Table 2,
Retinal penetration
Fig. 2). An oral dose of 500 mg/kg compound 1 The retinal drug levels were determined after
showed neuroprotective efficacy in the rat retinal oral administration of 1, 3, 5, and 6. The results
ischemia model. The oral dose was selected to
compare the potential efficacy of these com-
pounds. Aldehyde 1 had a very high rate of total
body clearance (Cl  89 mL/min/kg), which ex-
ceeds the hepatic blood flow for a rat (approxi-
mately 60 mL/min/kg), resulting in a short half-
life, in spite of its large volume of distribution at
steady state (Vss) of 6.1 L/kg. Compounds 3, 5,
and 6 are well absorbed and showed significantly
higher oral BA than original aldehyde 1. The Tmax
of 3, 5, and 6 were shorter (Tmax  0.25 h) than
that of 1. After i.v. dosing, 3, 5, and 6 showed
longer plasma terminal half-life (T1/2) and lower
total clearance (Cl), compared to 1. Hemiacetal 3
demonstrated relatively higher clearance (Cl  55
mL/min/kg), compared to the value estimated
by its excellent in vitro metabolic stability (Table
1). Ketoamide 5 exhibited excellent BA (F  96%)
FIG. 2. Plasma levels of aldehyde 1 (500 mg/kg), hemi-
and the longest half-life (T1/2  4.3 h) among acetal 3, amphipathic ketoamide 5, and pyridine ke-
these compounds. Although the structural dif- toamide 6 after oral administration to rats (10 mg/kg).
ference between ketoamides 5 and 6 was the only Results represent mean  standard deviation (n  4–5).
422 SHIRASAKI ET AL.

ratio in these compounds, whereas it had the

shortest retinal half-life (t1/2  2.8 h). Ketoamide
5 exhibited moderate Cmax (0.40 M), high AUC
(2.4 M  h), C8h (0.11 M), and R/P ratio (0.32).
In addition, 5 displayed the longest t1/2 (4.3 h) as
well as plasma half-life (T1/2). Pyridine ke-
toamide 6 showed relatively low AUC and R/P
ratio, whereas the t1/2 was moderate (2.9 h). The
elimination rate constant (z) in retina of 6 was
larger than that of 5, in spite of the larger
lipophilicity of 6. Among these compounds, ke-
toamide 5 exhibited more favorable retinal PK
properties, such as good penetration, long half-
life, and high C8h.

FIG. 3. Retinal levels of aldehyde 1 (500 mg/kg), hemi-

acetal 3, amphipathic ketoamide 5, and pyridine ke-
toamide 6 after oral administration to rats (10 mg/kg).
Results represent mean  standard deviation. (n  4–5).
were summarized in Figure 3 and Table 3. After
oral administration of these compounds, the Cmax
in plasma and retinas occurred simultaneously
and the retinal half-lives (t1/2) roughly reflected
their plasma T1/2 values. The retina-to-plasma
AUC ratios (R/P ratio) of 3, 5, and 6 were signif-
icantly higher than that of original aldehyde 1.
The retinal Cmax, AUC, and concentration at 8 h
after oral administration (C8h) of 3, 5, and 6 were
slightly less or comparable to those of 1, although
the oral doses of 3, 5, and 6 were fiftyfold less
than that of 1. Hemiacetal 3 showed the highest
Cmax (1.8 M), AUC, (2.6 Mh), and R/P (0.46)
DISCUSSION
Original aldehyde 1 showed very poor retinal
AUC and R/P ratio (0.062), although it exhib-
ited low molecular weight (Mw  372) and a
high Vss value (Vss  6.1 L/kg). It was suggested
that the aldehyde moiety of 1 formed a covalent
adduct with nucleophiles contained in the cir-
culatory system. Furthermore, relatively high
lipophilicity (cLogD7 3.21) may lead to an active
efflux by transporters such as P-glycoprotein (P-
gp) of retinal pigmented epithelium.27 Both the
reactive aldehyde moiety and high lipophilic na-
ture may be the limiting factors of retinal pene-
tration.
Hemiacetal 3 and ketoamide 5 and 6 exhibited
both higher oral BA and retinal penetration com-

TABLE 3. RETINAL PK PARAMETERS AFTER SINGLE ORAL ADMINISTRATION

(1500 MG/KG; 3, 5 AND 6: 10 MG/KG) OF CALPAIN INHIBITORS IN RATS

Parameters 1 3 5 6

R/P ratioa 0.062 0.46 0.32 0.10

AUCb (M  h) 2.8/(0.056)f 2.6 2.4 0.71
(g  h/mL) 1.0/(0.021)f 0.91 1.1 0.35
Cmax (M) 0.60/(0.012)f 1.8 0.40 0.51
(g/mL) 0.22/(0.045)f 0.63 0.20 0.25
Tmax (h) 1.0 0.25 0.25 0.25
T1/2c (h) 2.8 2.3 4.3 2.9
zd (h1) 0.25 0.30 0.16 0.24
C8he (M) 0.10/(0.0021)f 0.050 0.11 0.021
(g/mL) 0.037/(0.00078)f 0.018 0.054 0.010

PK, pharmacokinetic; R/P, retina to plasma; AUC, areas under the curve.
aRetina-to-plasma AUC ratio.
bThe area under the curves for 0 h to infinity.
cThe terminal half-life.
dThe terminal phase elimination rate constant.
eThe retinal concentration at 8 h after administration.
fThe value dose-normalized to 10 mg/kg.
RETINAL PENETRATION OF CALPAIN INHIBITORS
pared to 1, owing to the presence of a more chem-
ically stable warhead instead of aldehyde. Hemi-
acetal 3 showed the highest retinal AUC, R/P ra-
tio, and Cmax values, although the oral BA and
Vss were lower than those of 5. This is most likely
the result of a smaller molecular weight and
lower lipophilicity, which leads to weaker plasma
protein binding and an increase in unbound frac-
tions in plasma. The relatively high plasma clear-
ance of 3 may be ascribed to low plasma protein
bindings and metabolism by plasma esterases.
Hemiacetal 3 demonstrated a shorter retinal half-
life (t1/2), presumably owing to its high mem-
brane permeability and low lipophilic nature.
Among these compounds, ketoamide 5 showed
the most favorable PK profile. The retinal AUC
value of 5 was approximately equal to that of
hemiacetal 3. Furthermore, ketoamide 5 demon-
strated the longest retinal half-life (t1/2) and the
highest concentration at 8 h after oral adminis-
tration (C8h). Ketoamide 5 incorporates methoxy
diethylene glycol (mDEG) moiety as a nonionic
amphiphile at the P3 site and exhibits adequate
lipophilicity. On the other hand, structurally re-
lated ketoamide 6 has a pyridineethanol as a ba-
sic water-soluble moiety at the P3 site and pos-
sesses a similar lipophilicity to 1. It has been
reported that the introduction of a basic moiety
led to an increase in plasma half-life and volume
of distribution through a charge-charge interac-
tion between the protonated base and a region of
negative charge in the membrane phospho-
lipids.30 However, incorporation of basic pyri-
dine moiety resulted in lower Vss, retinal pene-
tration, and larger, shorter half-life. These
unfavorable results may be caused by strong
plasma protein binding owing to its high
lipophilic nature, leading to active elimination
from the retina by efflux transporters in BRB,
such as P-gp.27 Comparisons of ketoamides 5 and
6 revealed that mDEG moiety at the P3 site of 5
played a crucial role for well penetrating into the
retina and sustaining the high retinal drug levels
through the addition of the adequate lipophilic-
ity to the dipeptidyl ketoamide backbone.
This study disclosed that orally bioavailable in-
hibitors 3, 5, and 6 showed higher retinal pene-
tration than original aldehyde 1. Compound 1
was detected at levels 0.1–0.6 M in the retina fol-
lowing an oral dose of 500 mg/kg, which is the
minimum effective dose in the rat retinal is-
chemia model. The potencies of these inhibitors
were comparable to that of 1 (Table 1). If the reti-
423
nal levels exceeded 0.1 M at an oral dose of 10
mg/kg, the effective oral dose in the ischemia
model could be an order of magnitude smaller
than that of 1. On the other hand, the mean reti-
nal levels of 3 and 5 were 0.06–1.8 and 0.11–0.40
M, respectively, after oral dosing of 10 mg/kg.
These indicated that the compounds would pro-
vide retinal efficacy at a significantly lower oral
dosage, compared to 1. In fact, 3 and 5 demon-
strated neuroprotective efficacy at a fivefold
lower dose (100 mg/kg) in the rat retinal ischemia
model,23,31 although these were not effective at a
dose of 30 mg/kg. The ketoamide 5 exhibited the
longest t1/2 value and the highest concentration
at 8 h after oral administration (C8h  0.11 M)
among these compounds. Therefore, 5 is the most
favorable compound as a candidate for the treat-
ment of chronic retinal diseases.
CONCLUSIONS
In conclusion, we evaluated pharmacokinetic
properties in the retina after oral administration
of the several classes of calpain inhibitors, hemi-
acetal 3, ketoamide 5, and ketoamide 6, to obtain
an oral drug for the treatment of retinal disorders.
This study confirmed that these compounds have
remarkably higher penetration into the retina af-
ter oral administration than original aldehyde 1.
Ketoamide 5, containing an amphipathic moiety,
had excellent oral BA, high retinal penetration,
and a long retinal half-life. Comparisons of 5 with
pyridine ketoamide 6 revealed that the mDEG
moiety at the P3 provided the good PK proper-
ties in plasma and retinas. Ketoamide 5 would
show the retinal efficacy in various retinal disor-
ders in lower oral dosage, compared to 1 and a
long duration of action. In future work, we will
assess ketoamide 5 as the candidate for further
development in adequate pharmacological mod-
els, which reflect retinal diseases.
ACKNOWLEDGMENTS
The authors would like to thank Drs. Yukuo
Yoshida and Mitsuyoshi Azuma for reading the
manuscript. In addition, we thank Noriko Nada
for supporting laboratory work. Also, we would
like to thank Drs. Yutaka Kawamatsu, Jun Inoue,
and Masayuki Nakamura for useful discussions.
424
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Received: April 20, 2006
Accepted: July 19, 2006
Reprint Requests: Yoshihisa Shirasaki
Pharmacokinetic Group
Laboratory for Preclinical Research
Senju Pharmaceutical Co., Ltd.
1-5-4 Murotani, Nishiku
Kobe, Hyogo 651-2241
Japan
E-mail: shirasaki@senju.co.jp