Title of Invention

PARTICLE-FORMING COMPOSITIONS CONTAINING FUSED PYRROLOCARBAZOLES

Abstract A composition comprising a fused pyrrolocarbazole with the formula: wherein R3 and R4 are selected from H, a straight chain, cyclic or branched alkyl group having 1 to 8 carbon atoms, CI, Br, CH2OH, CH2SOCH2CH3, CH2SO2CH2CH3 NHCONHC6H5, CH2SCH2CH3, CH2SC6H5, NHCO2CH3, CH2OC(=O)NHCH2CH3, N(CH3)2, CH=NNH, CH2N(CH3)2 and CH2OCH2CH3; R7 is selected from H and alkyl; and R15 and R16 are independently selected from H, alkyl, OH, CH2OH, alkoxy, and CO2alkyl; or a stereoisomer or pharmaceutically acceptable salt form thereof; at least 20% (w/w) of a poloxyl stearate; and at least one polyethylene glycol.
Full Text


FIELD OF THE INVENTION
The invention relates to non-aqueous, particle-
forming pharmaceutical compositions containing fused
pyrrolocarbazoles or derivatives thereof. The invention
is also directed to stable solutions which result upon
contacting the particle-forming compositions of the
present invention with an aqueous, medium. The invention
is further directed to methods of medical treatment which
include treatment of a subject with the compositions or
solutions of the present invention.
BACKGROUND OF THE INVENTION
Fused pyrrolocarbazoles display various
pharmacological activities. For example, fused
pyrrolocarbazoles are useful for treatment of neurological
diseases or disorders, and some display antifungal,
antimicrobial, or antitumor activity. In some cases this
is accomplished by modulation of neurotrophic responses
through effects on protein kinase activity (Berg et al.
J. Biol. Chem. 267:13-16 (1992)). Fused carbazoles and
their derivatives have been isolated from a various
microorganisms, including S. staurosporeus, N.
aerocoligenes, Actinomadura and Nocardiopsis sp, (Kase et
al. , Biochem. Biophys. Res. Commun. 142: 436-440,
(1987)).
Specific fused pyrrolocarbazoles, such as
indolocarbazoles, which have been characterized include
the following: staurosporine and rebeccamycin (Moody et
al., supra); K-252a, K-252b (Kase et al., supra); K-252c

(also called staurosporine aglycon) (Moody et al., supra),
K-252d and derivatives thereof (published Japanese patent
applications 60-257652, 60-295172, 62-327858, 62-327859,
and 60-295173). K-252a, K-252b, K-252c, and K-252d are
insoluble in water (Nakanishi et al. , J. Antibodies,
34:1066, (1986)).
In general, fused pyrrolocarbazoles display very
low water solubility. Dry pharmaceutical preparations
(dragees, tablets and capsules) of staurosporine
derivatives that may contain polyethylene glycol and
polyvinylpyrrolidone have been described in U.S. Patent
No.. 5,093,330. Conventional pharmaceutical formulations
that include indolocarbazoles are described in U.S. Patent
No. 5,043,335 and PCT publication No. WO 93/00909.
Aqueous indolocarbazole compositions are described in U.S.
Patent No. 5,599,808.
Self-emulsifying drug delivery systems ("SEDDS")
have been developed for drugs that display very low water
solubility but good oil solubility. See, e.g., Shah et
al.-., International Journal of Pharmaceutics (Netherlands)
106:15-23, (1994). Despite their low water solubility,
fused carbazoles are generally unsuited for SEDDS because
of their low oil solubility. .
SUMMARY OF THE INVENTION
Accordingly, one object of the present invention is
to provide particle-forming compositions containing fused
pyrrolocarbazoles or derivatives thereof. Particularly,
the compositions of the present invention are non-aqueous
and contain an amount of surfactant sufficient to make a
fused pyrrolocarbazole or a derivative thereof particle-
forming.
It is another object of the invention to provide a
stable suspension that comprises suspended fused

pyrrolocarbazole-containing particles that greatly
improves bioavailability upon oral administration.
It is another object of the invention to provide a
method of forming a stable suspension of fused
pyrrolocarbazole-containing particles which comprises
contacting the particle-forming compositions of the
present invention with an aqueous medium.
It is another object of the invention to provide a
method of treating a disease or disorder in a mammal which
comprises administering a therapeutically effective amount
of the particle-forming compositions of the present
invention.
These and other objects, which will become apparent
during the following detailed.description, have been
achieved by the inventor's discovery that fused
pyrrolocarbazoles and derivatives thereof can be
formulated with suitable concentrations of one or more
surfactants to make a non-aqueous, particle-forming
composition, wherein the composition spontaneously
disperses into suspended particles upon contact with an
aqueous medium.
DETAILED DESCRIPTION OF THE INVENTION
Thus, in a first embodiment, the present invention
provides a non-aqueous, particle-forming composition
comprising a fused pyrrolocarbazole and a surfactant. In
a preferred embodiment, the fused pyrrolocarbazole has
Formula I:


or a stereoisomer or pharmaceutically acceptable salt form
thereof, wherein:
ring B and ring F, are independently selected from:
a) an unsaturated 6-membered carbocyclic aromatic
ring in which from 1 to 3 carbon atoms may be
replaced by nitrogen atoms;
b) an unsaturated 5-membered carbocyclic aromatic
ring; and
c) an unsaturated 5-membered carbocyclic aromatic
ring in which either:

1) one carbon atom is replaced with an oxygen,
nitrogen, or sulfur;
2) two carbon atoms are replaced with a sulfur
and a nitrogen, an oxygen and a nitrogen,
or two nitrogens; or
3) three carbon atoms are replaced with three
nitrogens;

Z1 and Z2, at each occurrence, are independently selected
from H, H; H, OR; H, SR; H, N(R)2; and a group wherein
Z1 and Z2 together form a moiety selected from =0, =S,
and =NR; with the proviso that at least one of the

pairs Z1 and Z2 form =0;
R is selected from H, substituted or unsubstituted alkyl
having from 1 to 6 carbons, OH, alkoxy having from 1
to 4 carbons, 0C(=O)Rla, 0C (=O)NRlcRld, O (CH2)pNRlcRld,
O(CH2)p O Rlb, substituted or unsubstituted arylalkyl
having from 6 to 10 carbons, and substituted or
unsubstituted heteroarylalkyl;
R1 is selected independently from:
a) H, substituted or unsubstituted alkyl having froi
1 to 6 carbons, substituted or unsubstituted
aryl,
substituted or unsubstituted arylalkyl,
substituted
or unsubstituted heteroaryl, and substituted or
unsubstituted heteroarylalkyl;
b) C(=O)Rla;
c) 0Rlb;
d) C(=O)NHRlb, NRlcRld, (CH2)PNRlcRld, (CH2)pORlb,
O(CH2)P0Rlb and O (CH2)pNRlcRld;
Rla is selected from substituted or unsubstituted alkyl,
substituted or unsubstituted aryl and heteroaryl;
Rlb is selected from H and substituted or unsubstituted
alkyl
having from 1 to 6 carbons;
Rlc and Rld are each independently selected from H,
substituted or unsubstituted alkyl having from 1 to 4
carbons, and a linking group of the formula
-(CE2)2-X1-(CE2)2-;
X1 is selected from -0-, -S-, and -CH2-;
R2 is selected from H, SO2R2a, CO2R2a, C(=O)R2af C (=O) NR2cR2d,
and alkyl of 1-8 carbons, alkenyl of 2-8 carbons,
alkynyl of 2-8 carbons, wherein:
1) each alkyl, alkenyl, and alkynyl is
unsubstituted; or
2) each alkyl, alkenyl, and alkynyl is

substituted with 1-3 R5;
R2a is selected from alkyl of 1 to 6 carbons, aryl, OR2b,
CONH2, NR2cR2d, (CH2)pNR2cR2d, and 0 (CH2)pNR2cR2d;
R2b is selected from H and substituted or unsubstituted
alkyl
having from 1 to 6 carbons;
R2c and R2d are each independently selected from H,
substituted or unsubstituted alkyl having from 1 to 6
carbons, and a linking group of the formula
-(CH2)2-X1-(CH2)2-;
R3 and R4, at each occurrence, are independently selected
from:
a) H, aryl, heteroaryl, F, Cl, Br, I, CN, CF3, N02,
OH, OR9, O(CH2)pNR11R12 , OC(=O)R9, OC (=O)NR11R12,
0(CH2)pOR10, CH2OR10, NR1:1R12, NR10S (=O) 2R9, and
NR10C(=O)R9;
b) CH2OR14;
c) NR10C(=O)NR11R12, C02R10, C(=O)R9, C(=O)NRUR12,
CH=NOR10, CH=NR10, (CH2)pNR11R12, (CH2)pNHR14, and
CH=NNR11R12 ;
d) S(O)yR9, (CH2)pS(O)yR9 , CH2S(O)yR14;
e) alkyl having from 1 to 8 carbons, alkenyl having
from 2 to 8 carbons, and alkynyl having 2 to 8
carbons, wherein

1) each alkyl, alkenyl, or alkynyl group is
unsubstituted; or
2) each alkyl, alkenyl, or alkynyl group is
substituted with 1 to 3 R5;
R5 is selected from aryl having from 6 to 10 carbons,
heteroaryl, arylalkoxy, heterocycloalkoxy,
hydroxyalkoxy, alkyloxy-alkoxy, hydroxyalkylthio,
alkoxy-alkylthio, F, Cl, Br, I, CN, NO2, OH, OR9,
X2 (CH2) pNR11R12, X2 (CH2) PC (=O) NR11R12,
X2(CH2)pOC(=O)NR11R12, X2(CH2)pCO2R9, X2 (CH2)pS (O) yR9,
X2(CH2)PNR10C(=O)NR11R12, OC(=O)R9, OC(=O)NHR10,

O-tetrahydropyranyl, NR11R12, NR10C(=O)R9, NR10CO2R9,
NR10C(=O)NR11R12, NHC(=NH)NH2/ NR10S(O)2R9, S(O)yR9,
C02R10, C(=O)NR11R12/ C(=O)R9, CH2OR10, CH=NNR11R12,
CH=NOR10, CH=NR9, CH=NNHCH (N=NH)NH2, S (=O) 2NR11R12,
P(=O) (OR10)2, OR14, and a monosaccharide having from 5
to 7 carbons wherein each hydroxyl group of the
monosaccharide is independently either unsubstituted
or is replaced by H, alkyl having from 1 to 4
carbons, alkylcarbonyloxy having from 2 to 5 carbons,
or alkoxy having from of 1 to 4 carbons;
X2 is 0, S, or NR10;
Q is selected from:
1) NR6,
2) an unsubstituted alkylene of 1-3 carbons;
3) a substituted alkylene of 1-3 carbons;'
4) CH=CH, CH(0H)CH(0H), O, S, S{=0), S(=O)2, C{=0),
C(=NOR11), C(0R11) (R11) , C(=O)CH(R13), CH(R13)C (=O) ,
C(R10)2, C(=N0R11)CH(R13) , CH(R13)C (=NOR11) , CH2Z,
Z-CH2, CH2ZCH2 ;
Z is selected from C (R11) (OR11) , 0, S, C(=O), C(=NOR11), and
NR11;
R6 is selected from H, SO2R2a, C02R2a, C(=O)R2a, C (=O) NRlcRld,
and alkyl of 1-8 carbons, alkenyl of 2-8 carbons,
alkynyl of 2-8 carbons, wherein:
1) each alkyl, alkenyl, and alkynyl is
unsubstituted;
2) each alkyl, alkenyl, and alkynyl- is
substituted with 1-3 R5; or r
alternatively, when Q is NR6 or C(R10)2, R6 or one R10 joins
with R2 to form:


wherein R7 and R8 are each independently selected from H,
OH,
alkyl having from 1 to 6 carbons, alkoxy having from
1 to 6 carbons, substituted or unsubstituted
arylalkyl having from 6 to 10 carbons, substituted or
unsubstituted heteroarylalkyl, (CH2)pOR10,

R7 and R8 together form a linking group of the formula
CH2-X3-CH2;
X3 is a bond, 0, S, or NR10;
R9 is selected from alkyl having 1 to 6 carbons, (CH2)rary3
and (CH2) rheteroary 1 ;
R10 is selected from H, alkyl having from 1 to 6 carbons,
(CH2)raryl and (CH2)rheteroaryl;
R11 and R12, at each occurrence, are independently selected
from:
1) H and substituted or unsubstituted alkyl having
from 1 to 6 carbons; or
2) R11 and R12 together form - (CH2) 2-X1-(CH2) 2-;
Y is selected from 0, S, N(R10) , N+(O-)(R10), N(OR10) , and
CH2 ;
J is selected from the group consisting of a bond, 0,
CH=CH, S, C(=O), CH(OR10), N(R10), N(OR10), CH(NR11R12),
C(=O)N(R17), N(Rl7)C(=O), N(S(O)yR9), N (S (O)yNR11R12) ,
N(C(=O)R17), C(R15R16), N+(O-)(R10), CH (OH) CH(OH) , and
CH(O(C=O)R9)CH(OC(=O)R9) ;
R13 is selected from alkyl having from 1 to 4 carbons,
aryl,
and arylalkyl having from 7 to 14 carbons;
R14 is the residue of an amino acid after the hydroxyl
group of the carboxyl group is removed;
R15 and R16, at each occurrence is selected from H, OH,
C(=O)R10, O(C=O)R9, alkyl-OH, and CO2R10;
R1 is selected from the group consisting of H, alkyl,
aryl, and heteroaryl;

m and n are independently selected from 0, 1, and 2;
p is independently selected from 1, 2, 3, and 4;
r is independently selected from 0, 1, and 2; and
y is independently selected from 0, 1 and 2.
In certain preferred embodiments, rings B and F are
phenyl. In other preferred embodiments, G-X-W is select
from CH2NR1C(=O)/ C(=O)NR1CH2, and C (=O)NRaC (=O) . In oth preferred embodiments, Q is NR6.
In other preferred embodiments, the fused
pyrrolocarbazole has the Formula II-a:

In certain more preferred embodiments, 21 and Z2 an
Ji^R3 and R4 are selected from H, alkyl, Cl, Br, CH2OH,
CH2SOCH2CH3/ CH2SO2CH2CH3, NHCONHC6H6, CH2SCH2CH3, CH2SC6H5,
•NHCO2CH3, CH2OC(=O)NHCH2CH3, N(CH3)2, CH=NNH, CH2N(CH3)2/
CH2OCH2CH3/ R7 is selected from H and alkyl, and R15 and R:
are independently selected from H, alkyl, OH, CH20H,
alkoxy, and CO2alkyl. In certain even more preferred
embodiments, the fused carbazoles are indolocarbazoles a:
set forth in Tables 1-A and 1-B.

In certain preferred embodiments, the particle-
orming composition further comprises one or more of an
rganic solvent, a lipid, or an antioxidant. In certain
referred embodiments, the amount of surfactant is at
least about 20% (w/w). In certain more preferred
embodiments, the amount of surfactant is at least 40%
about (w/w). In certain more preferred embodiments, the
amount of surfactant is at least about 50% (w/w).
In certain preferred embodiments, the surfactant is
selected from:polyethylene glycol stearate d-α-tocopheryl
polyethylene glycol succinate, poloxyl stearate, poloxyl
castor oil, a polyoxyethylene sorbitan fatty acid ester, a
polyethylene glycol ether, a saturated polyglycolized
glyceride, a fatty acid ester of polyethylene glycol, a
hydroxylated lecithin, a medium chain monoglyceride, a
medium chain fatty acid ester, and polyethylene-propylene
glycol copolymer. In other preferred embodiments, the
lipid is a diester of coconut fatty acids and[propylene
glycoll In other preferred embodiments, the organic
solvent is selected from propylene glycol, propylene
carbonate, dimethyl isosorbide, and polyethylene glycol
(PEG)u In other preferred embodiments, the antioxidant is
selected from ascorbic acid, a fatty acid ester of
ascorbic acid, and butylated hydroxyanisole.
In certain preferred embodiments, the fused
pyrrolocarbazole is present at a concentration of about 1
to about 300 mg/ml. In certain more preferred
embodiments, the fused pyrrolocarbazole is present at a
concentration of about 1 to about 100 mg/ml. In certain
further more preferred embodiments, the concentration is
about 1 to about 50 mg/ml.
In certain embodimencs, the composition is a semi-
solid or solid at room temperature. In certain more
preferred embodiments, the semi-solid or solid is in the
form of a capsule or to

In another embodiment, the present invention provides
a stable suspension of fused pyrrolocarbazole-containing
particles in an aqueous suspension medium. In certain
preferred embodiments, the particles have a diameter less
than about 400 run. In certain more preferred embodiments,
the-particles have a diameter less than about 100 nm.
In another embodiment, the present invention provides
a method of forming a stable suspension of fused
pyrrolocarbazole-containing, suspended particles,
comprising contacting a fused pyrrolocarbazole in a non-
aqueous liquid containing a surfactant in an amount from
about 20% to greater than 99% with an aqueous medium. In
a preferred method the particle-forming composition is
contacted with an aqueous medium in vitro. In another
preferred method, the particle-forming composition is
contacted with an aqueous medium in vivo.
In another embodiment, the present invention provides
; a method of forming a stable suspension of fused
pyrrolocarbazole-containing, suspended particles,
comprising:
(a) dissolving a fused pyrrolocarbazole in a non-
aqueous liquid containing a surfactant in an amount from
about 20% to greater than 99%, to form a particle-forming
composition; and
(b) contacting the particle-forming composition with
an aqueous medium to form a stable suspension.
In another embodiment, the present invention provides
a method of treating a disease or disorder in a mammal,
comprising administering a therapeutically effective
amount of a fused, pyrrolocarbazole in a non-aqueous,
particle-forming composition comprising a surfactant in an
amount of at least 20% (w/w) to the mammal.
In another embodiment, the present invention provides
a method of treating a disease or disorder in a mammal,


comprising:
(a) contacting a fused pyrrolocarbazole in a non-
aqueous, particle-forming composition comprising a
surfactant in an amount of at least 2 0% (w/w) with an
aqueous medium, thereby forming a stable suspension
comprising suspended particles; and
(b) administering a therapeutically effective amount
of the stable suspension to the mammal.
In certain preferred embodiments the surfactant is
present in at least 40% (w/w). In other preferred
embodiments, the disease or disorder is a neurological
disorder, or cancer such as prostate cancer. In other
preferred embodiments, the subject is a mammal.[J
The materials, metliods, and examples presented herein
are intended to be illustrative, and not intended to limit
the scope of the invention. All publications, patent
applications, patents, and other references mentioned
herein are incorporated by reference.• Unless otherwise
i
defined, all technical and scientific terms are intended
to have their art-recognized meanings.
The present invention is directed to non-aqueous,
particle-forming compositions which contain a fused
pyrrolocarbazole, and a surfactant. Upon contact with ai
aqueous medium, the particle-forming composition
spontaneously disperses into suspended particles, thereby
forming a stable suspension that provides greatly improve
bioavailability of orally administered fused
pyrrolocarbazole compounds.
The particle-forming compositions of the present
invention may be a liquid, semi-solid, or solid at room
J
temperature. If liquid, the composition may be contained
in a capsule. If semi-solid or solid, the composition can
be in the form of a capsule or tablet.
As used herein, "non-aqueous" composition is intended '
to mean a composition that contains from 0% to about 10%
water by weight. As used herein, "particle-forming
composition" is intended to mean a composition that
spontaneously disperses into suspended particles upon
entry into an aqueous medium. As used herein, "suspended
particle" is intended to mean a micelle, microsphere,
droplet, or other substantially non-crystalline physical
structure that remains suspended in an aqueous medium,
without substantial phase separation. As used herein,
"aqueous medium" is intended to mean any medium comprised
of greater than 10% water, and in which the compositions
of the present invention are particle-forming.
Initially, a fused pyrrolocarbazole is dissolved in a
non-aqueous,.particle-forming composition. Subsequently,
the particle-forming composition is contacted with an I
aqueous medium to form an aqueous suspension. Upon
contact with an aqueous medium, the fused
pyrrolocarbazole-containing, non-aqueous, particle-forming
composition spontaneously forms particles of a suitable
size, i.e., without energy input. Thus, when initially
dissblved in a non-aqueous, particle-forming composition,
a fused pyrrolocarbazole may have a higher solubility as
compared to its solubility when placed directly in a
comparable aqueous suspension containing preformed
particles.
The particle-forming composition can be contacted
with an aqueous medium in vitro, i.e., subjected to
predilution, prior to ingestion by a mammal.
Alternatively, the initial contact with an aqueous medium
can be in vivo, e.g., contact with aqueous contents of the
gastrointestinal composition of a mammal.
When the particle-forming composition is subjected to
predilution, the dilution ratio preferably is from about
1:1000 (1 part formulation to 999 parts aqueous medium) to
about 1:2 (1 part formulation to 1 part aqueous medium).

More preferably, the dilution ratio is from about 1:500 (1
part formulation to 499 parts aqueous medium) to about 1:3
(1 part formulation to 2 parts aqueous medium) . By way of
general guidance, for administration to humans a
convenient ratio is about 1:250, which is a rough
correspondence to a 1 ml unit dose dispersed in an 8-qunce
glass of an aqueous liquid.
It has also been discovered that the resultant
solution in which the fused pyrrolocarbazole-containing .
particles are suspended is a stable suspension.
Preferably, the particles contained in this medium have a
diameter less than 400 nm. More preferably, the particles
have a diameter less than 100 nm.
The degree of optical transparency of a given volume
of water containing a given amount of formulation gives a
useful indication of particle size. This is because the
particles scatter visible light, with the larger particles
causing greater scattering. In general, the greater the
optical transparency, the smaller the particle size. High
optical transparency, i.e., bluish haze invisible or
nearly invisible, generally indicates a particle size of
less than 100 nm. A distinct bluish haze generally
indicates a particle size from about 100 nm to about 400
nm. Without intending to be bound by theory, it is noted
that particle size tends to be essentially constant for a
given formulation, regardless of the dilution ratio. If
particles fail to form, an increase in dilution ratio may
be used to promote particle formation.
Whether a formulation according to the invention is a
liquid, semi-solid, or solid at room temperature, may
depend upon the selection of components, or other concerns
such as commercial viability, administration and the like.
For example, a semi-solid or solid formulation is
convenient for manufacturing unit doses of a fused
pyrrolocarbazole in the form of a capsule, including both

hard gelatin and soft gelatin capsules, and tablets. When
the liquid or solid formulation contacts an aqueous
medium, e.g., gastrointestinal liquids, the formulation
disperses into suspended particles in which the fused
pyrrolocarbazole is biologically available.
Compositions whose inert components (i.e., components
other than the fused pyrrolocarbazole) are all liquid at
room temperature can be prepared by simply mixing the
components without heating. The desired amount of fused
pyrrolocarbazole can be weighed out and dissolved in the
mixture of inert components, without heating. Moderate
heating, preferably less than 60°C, can be applied to
hasten complete mixing of the inert components, to hasten
dissolution of the fused pyrrolocarbazole, or both.
Preparation of compositions containing one or more
components that are solid at room temperature is carried
out at a moderately elevated temperature, preferably less
than 60°C. While moderate heating can be useful,
excessive heating can cause decomposition of one or more
components of the formulation. For example, decomposition
of polysorbate 80 can occur at temperatures above 60°C.
Instability of certain fused pyrrolocarbazoles has been
Observed in the presence of short chain polyethylene
glycols, e.g., PEG 400, at temperatures in the range of
about 60°C to 90°C.
Decomposition of polysorbate 80 may occur if
maintained at 90°C for more than one hour. As will be
appreciated by one those of ordinary skill in the art, any
deleterious effects of heat accumulate with time.
Therefore, when heat is applied, time and temperature will
typically be balanced against one another.
There is wide latitude in formulation of all
particle-forming compositions of the present invention.
Preferably, all non-fused pyrrolocarbazole components in
the particle-forming composition are food grade materials

or GRAS (Generally Recognized As Safe) materials.
Information on GRAS materials can be found in Inactive
Ingredient Guide, published by the U.S. Food and Drug
Administration (Division of Drug Information Resources,
Rockville, MD), the disclosure of which is hereby
incorporated herein by reference in its entirety.
.Inactive Ingredient Guide provides a listing of all
inactive ingredients present in approved or conditionally
approved drug products currently marketed for human use.
The compositions of the present invention, preferably
contain a surfactant. Although the surfactant may be
present in any amount which results in a particle-forming
composition, typical compositions contain from about 20%
to greater than 99% of a surfactant. Preferably, the
amount of surfactant is at least 30%, at least 40%, or at
least 50%, depending upon the additional components of the
composition.
Surfactants include, but are not limited to,
polyoxyethylene stearates, polyoxyethylene castor oil,
polyoxyethylene sorbitan fatty acid esters (sorbitans),
polyethylene glycol ethers, saturated polyglycolized
glycerides, fatty acid esters of polyethylene glycol,
tiydroxylated lecithins, medium chain monoglycerides,
medium chain fatty acid esters, polyethylene/propylene
glycol copolymers, polyethylene glycol stearate, d-α-
tocopheryl polyethylene glycol succinate, poloxyl stearate
(e.g., Myrj® 52) and poloxyl castor oil.
Polyoxyethylene sorbitan fatty acid esters
(polysorbates) are non-ionic surfactants (detergents) that
may consist of a mixture of fatty acids. Commercially
available examples are Tween® 2 0 (polyoxyethylene (20)
sorbitan monolaurate) , Tween® 40 (polyoxyethylene (20)
sorbitan monopalmitate) , and Tween® 80 (polyoxyethylene
(20) sorbitan monooleate).

A preferred polyethylene glycol ether is PEG-4-
isooctylphenyl ether, a non-ionic surfactant.
Commercially available examples of polyethylene glycol
ether surfactants are Triton® X-100, Triton® X-114, Triton®
X-405, and Triton® N-101. Non-ionic surfactants are
preferred.
Examples of other useful surfactants are saturated
polyglycolized glycerides consisting of mono-, di-, or
triglycerides; di-fatty acid esters of polyethylene
glycol, e.g., Gelucire® 44/14; hydroxylated lecithins,
e.g., Centrolene® A; medium chain monoglycerides, e.g.,
glyceryl monocaprylate (Imwitor® 308, Capmul® MCM C-8);
medium chain monoglycerides and diglycerides, e.g.,
glyceryl caprylate/caprate (Capmul® MCM);
polyethylene/propylene glycol copolymers; block copolymers
of ethylene oxide and propylene oxide (e.g., Poloxamer
188, Pluronic® F-68); ethoxylated castor oil (e.g.,
Cremophor® EL); and ethoxylated hydroxystearic acid (e.g.,
Solutol® HS 15) . Some surfactants are solid or semisolid
at room temperature, e.g., Poloxamer•188, glyceryl
monocaprylate, and Gelucire® 44/14. Additional
surfactants are those found in The Handbook of
Pharmaceutical Excipients, 2nd Ed., published by The
Pharmaceutical Press, London and American Pharmaceutical
Association (1994) , a common text in the field, which is
hereby incorporated by reference in its entirety.
The particle-forming compositions may also include an
organic solvent, a lipid, an antioxidant, or a combination
of these components. Such components include those found
in The Handbook of Pharmaceutical Excipients, and include
any which are known in the art to be acceptable for use in
pharmaceutical formulations. The selection of suitable
components is within ordinary skill in the art.
In some embodiments of the invention, inclusion of an

organic solvent improves solubility of the fused
pyrrolocarbazoles in the particle-forming composition.
Certain useful organic solvents form a solid (or semi-
solid) at room temperature, e.g., polyethylene glycol
1450. The effectiveness of an organic solvent may depend,
in part, on the particular fused pyrrolocarbazole included
in the particle-forming composition. By way of general
guidance, organic solvents include, but are not limited
to, propylene carbonate, dimethyl isosorbide, benzyl
alcohol, and glycols such as propylene glycol and
polyethylene glycol (PEG). As used herein, "polyethylene
glycol" or "PEG" means a liquid
or solid polymer "of the general formula H(OCH2CH2)nOH,
wherein n is at least 4.
In some embodiments of the invention, a suitable
antioxidant is included as a component of the particle-
forming composition. As used herein, "antioxidant" is
intended to indicate any substance useful to retard
deterioration by oxidation or to inhibit reactions
promoted by oxygen or peroxides. Antioxidants include,
but are not limited to, ascorbic acid; fatty acid esters
of ascorbic acid, e.g., ascorbyl palmitate; butylated
hydroxytoluene ("BHT"); propyl gallate; and butylated
hydroxyanisole ("BHA"). An appropriate antioxidant also
includes mixtures of antioxidants, such as a mixture of
BHA, propyl gallate and citric acid. An antioxidant is
"particularly useful when the fused pyrrolocarbazole
contains a functional group susceptible to oxidation,
e.g., a thiol or a thioether. A specific example of such
a fused pyrrolocarbazole is the indolocarbazole compound
IIa-51, which contains two thioether functionalities. An
antioxidant can function either by scavenging oxidative
compounds or inhibiting oxidation reactions.
In some embodiments of the invention, a suitable
lipid is included as a component of the particle-forming

composition. As used herein, "lipid" is intended to
indicate a fat, oil, wax, sterol, glycerol ether,
triglyceride, or combination thereof. The inclusion of a
lipid can change particle characteristics, including
particle size. One way in which a lipid can change
particle characteristics is by causing formation of a
microemulsion rather than micelles. Lipid-induced changes
in particle characteristics can also affect
bioavailability.
Although the compositions described herein may
conceivably employ any therapeutic substance with poor
aqueous solubility characteristics, the particle-forming
compositions preferably contain a fused pyrrolocarbazole.
Preferably the fused pyrrolocarbazole is present in the
particle-forming composition at a concentration of 1 to
300 mg/ml. More preferably, the fused pyrrolocarbazole is
present at a concentration of 1 to 100 mg/ml. Even more
preferably, the fused pyrrolocarbazole is present at a
concentration of about 1 to 50 mg/ml.
'As used herein, "fused pyrrolocarbazole" is intended
to mean a compound having a fused pyrrolocarbazole core '
structure:

wherein at least one of G, X, or W is a nitrogen, B is an
aryl or heteroaryl group, and * indicates the attachment
point of an additional fused ring system.
The core structures provided herein are presented by
way of general guidance, and are not to be taken as
limiting the scope of the invention. For example, certain

cores indicate the presence certain atoms for illustratve
purposes. It will be appreciated that such atoms may
bonded to additional groups, or may be further substitute
without deviating from the spirit of the invention.
Thus, pyrrolocarbazole core structures include, but
are-not limited to, structures of formula la:

wherein Q may be a moiety containing one or more nitroger
or carbons. Such structures include, but are not limited
to, indolocarbazoles, indenocarbazoles, and bridged
indenocarbazoles.
As used herein, "indolocarbazole" is intended to
indicate a compound of formula la, wherein Q is nitrogen
and Y is a bond:

These compounds are intended to include, but are not
limited to, structures in which the nitrogens of the
carbazole and the indole form a cyclic bridged moie,ty:


Such bridged structures include, but are not limited
to, derivatives of the microbial-derived material referred
to as K-252a.
As used herein, "indenocarbazole" is intended to
indicate a compound of formula la in which Q is not
nitrogen. These compounds include, but are not limited
to, compounds wherein Q is one or more carbons. For
example, in certain indenocarbazoles, Q is a single
carbon:

As used herein, "bridged indenopyrrolocarbazole" is
intended to indicate a compound of formula la in which Q
is a moiety containing at least one carbon which joins
with the nitrogen of the carbazole derivative to form a
bridged moiety:


particle-forming compositions of the present invention are
stable compounds. As used herein "stable compound" or
"stable structure" is meant to indicate a compound that is
sufficiently robust to survive isolation to a useful
degree of purity from a reaction mixture, and preferably
capable of,formulation into an efficacious therapeutic
agent.
The fused pyrrolocarbazoles may be further
substituted. As used herein, "substituted" is intended to
indicate that one or more hydrogen atoms on the indicated •
atom is replaced with a selected group referred to herein
as a "substituent", provided that the substituted atom's
valency is not exceeded, and that the substitution results
in a stable compound.
As used herein, the term "alkyl" means a straight-
chain, cyclic, or branched alkyl group having 1 to 8
carbon atoms, such as methyl, ethyl, propyl, isopropyl,
butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isoamyl,
neopentyl, 1-ethylpropyl, hexyl, octyl, cyclopropyl, and
cyclopentyl. The alkyl moiety of alkyl-containing groups,
such as alkoxy, alkoxycarbonyl, and alkylaminocarbonyl
groups, has the same meaning as alkyl defined above.
Lower alkyl groups, which are preferred, are alkyl groups
as defined above which contain 1 to 4 carbons.
Alkyl groups and alkyl moieties contained within
substituent groups such as aralkyl, alkoxy, arylalkoxy,
hydroxyalkoxy, alkoxy-alkoxy, hydroxy-alkylthio, alkoxy-
alkylthio, alkylcarbonyloxy, hydroxyalkyl and acyloxy
groups may be substituted or unsubstituted. A substituted
alkyl group has 1 to 3 independently-selected
substituents, preferably hydroxy, lower alkoxy, lower
alkoxy-alkoxy, substituted or unsubstituted arylalkoxy-
lower alkoxy, substituted or unsubstituted

heteroarylalkoxy-lower alkoxy, substituted or
unsubstituted arylalkoxy, substituted or unsubstituted
heterocycloalkoxy, halogen, carboxyl, lower
alkoxycarbonyl, nitro, amino, mono- or di-lower
alkylamino, dioxolane, dioxane, dithiolane, dithione,
furan, lactone, or lactam.
As used herein, the term "alkenyl" is intended to
include straight-chain, cyclic, or branched hydrocarbon
chains having at least one carbon-carbon double bond.
Examples of alkenyl groups include ethenyl, propenyl, 3-
methylbutenyl, and cyclohexenyl groups. As used herein,
the term' "alkynyl" is intended to include straight-chain,
cyclic, or branched hydrocarbon chains having at least one
carbon-carbon triple bond. Examples of alkynyl groups
include ethynyl, propynyl, 3-methylbutynyl, and
cyclohexynyl groups.
As used herein, the "acyl" moiety of acyl-containing
groups such as acyloxy groups is intended to include a
straight-chain, branched, or cyclic alkanoyl group having
1 to 6 carbon atoms, such as formyl, acetyl, propanoyl,
butyryl, valeryl, pivaloyl or hexanoyl.
As used herein, the term "carbocyclic" refers to
cyclic groups in which the ring portion is composed solely
of carbon atoms. These include, but are not limited to,
cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,
cycloheptyl, cyclooctyl. The term "carbocyclic aromatic
ring" is intended to include carbocyclic rings which are
also aryl rings. The terms "heterocyclo" and
"heterocyclic" refer to cyclic groups in which the ring
portion includes at least one heteroatom such as 0, N, or
S. Heterocyclyl groups include heteroaryl and heteroalkyl
groups.
As used herein the term "aryl" means an aromatic ring
having 6 to 12 carbon atoms such as phenyl, biphenyl and
naphthyl. Preferred aryl groups include unsubstituted or

substituted phenyl and naphthyl groups. The term
"heteroaryl""'as used herein denotes an aryl group in whic
one or more ring carbon atoms is replaced by a hetero
(i.e., non-carbon) atom such as 0, N or S. Preferred
heteroaryl groups include pyridyl, pyrimidyl, pyrrolyl,
furyl, thienyl, imidazolyl, triazolyl, tetrazolyl,
quinolyl, isoquinolyl, benzoimidazolyl, thiazolyl,
pyrazolyl, and benzothiazolyl groups. The term
"heteroalkyl" denotes a cycloalkyl group in which one or
more ring carbon atoms is replaced by hetero atoms such as
0, N, or S.
As used herein, the term "aralkyl" (or "arylalkyl")
is intended to denote a group having from 7 to 15 carbons,
consisting of an alkyl group that bears an aryl group.
Examples of aralkyl groups include, but are not limited
to, benzyl, phenethyl, benzhydryl and naphthylmethyl
groups. Substituted aryl, substituted heterocyclic and
substituted aralkyl groups each have 1 to 3 independently
selected substituents that are preferably lower alkyl,
hydroxy, lower alkoxy, carboxy, lower alkoxycarbonyl,
nitro, amino, mono- or di-lower alkylamino, and halogen.
Preferred heterocyclic groups formed with a nitrogen
atom include pyrrolidinyl, piperidinyl, piperidino,
morpholinyl, morpholino, thiomorpholino, N-
methylpiperazinyl, indolyl, isoindolyl, imidazole,
imidazoline, oxazoline, oxazole, triazole, thiazoline,
thiazole, isothiazole, thiadiazoles, triazines, isoxazole,
oxindole, indoxyl, pyrazole, pyrazolone, pyrimidine,
pyrazine, quinoline, iosquinoline, and tetrazole groups.
Preferred heterocyclic groups formed with an oxygen atom
include furan, tetrahydrofuran, pyran, benzofurans,
isobenzofurans, and tetrahydropyran groups. Preferred
heterocyclic groups formed with a sulfur atom include
thiophene, thianaphthene, tetrahydrothiophene,
tetrahydrothiapyran, and benzothiophenes.

As used herein, "hydroxyalkyl" groups are alkyl
groups that have a hydroxyl group appended thereto. As
used herein, "hydroxyalkoxy" groups are alkoxy groups that
have a hydroxyl group appended thereto. As used herein,
"halogen" refers to fluorine, chlorine, bromine and
iodine.
As used herein, the term "heteroarylalkyl" means an
arylalkyl group that contains a heteroatom in the aryl
moiety. The term "oxy" denotes the presence of an oxygen
atom.• Thus, "alkoxy" groups are alkyl groups that are
attached through an oxygen atom, and "carbonyloxy" groups
are carbonyl groups that are attached through an oxygen
atom.
As used herein, the terms "heterocycloalkyl" and
"heterocycloalkoxy" mean an alkyl or an alkoxy group that
has a heterocyclo group attached to the alkyl moiety
thereof, and the term "arylalkoxy" means an alkoxy group
that has an aryl group attached to the alkyl moiety
thereof. As used herein, the term "alkyIcarbony1oxy"
means a group of formula -0-CM=0)-alkyl.
As used herein, the term "alkyloxy-alkoxy" denotes an
alkoxy group that contains an alkyloxy substituent
. attached to its alkyl moiety. The term "alkoxy-alkylthio"
means an alkylthip group (i.e., a group of formula -S-
alkyl) that contains an alkoxy substituent attached to its
alkyl moiety. The term "hydroxy-alkylthio" means an
alkylthio group (i.e., a group of formula -S-alkyl) that
contains a hydroxy substituent attached to its alkyl
moiety.
As used herein, the term "monosaccharide" has its
accustomed meaning as a simple sugar. As used herein, the
term "amino acid" denotes a molecule containing both an
amino group and a carboxyl group. Embodiments of amino
acids include a-amino acids; i.e., carboxylic acids of
general formula H00C-CH(NH2)-(side chain). Side chains of

amino acids include naturally occurring and non-naturally
occurring moieties. Non-naturally occurring
(i.e., unnatural) amino acid side chains are moieties that
are used in place of naturally occurring amino acid side
chains in, for example, amino acid analogs. See, for
example, Lehninger, Biochemistry, Second Edition, Worth
Publishers, Inc, 1975, pages 73-75, incorporated by
reference herein. In certain embodiments, substituent
groups for the compounds described herein include the
residue of an amino acid after removal of the hydroxyl
moiety of the carboxyl group thereof; i.e., groups of
Formula -C (=O)CH(NH2) - (side chain).
The fused pyrrolocarbazoles are preferably present in
the compositions described herein in a therapeutically
effective amount. As used herein, a "therapeutically
effective'amount" refers to an amount of compound
effective to prevent or treat the symptoms of particular
disorder. Such disorders include, but are not limited to,
those pathological and neurological disorders associated
with the aberrant activity of target receptors, wherein
the treatment or prevention comprises inhibiting,
inducing, or enhancing the activity thereof by contacting
the receptor with an active drug substance. As will be
readily understood, the concentration and dosages of the
fused pyrrolocarbazoles will depend upon factors such as
the total dosage of the drug to be administered, the
chemical characteristics of the compounds employed, the
route of administration, the age, body weight and symptoms
of the patient, etc. By way of general guidance, human
doses may range from about 0.1 mg to about 1000 mg
administered per day. Preferably, the dosage is about 1
to about 500 mg administered two times a day. Even more
preferably, the dosage is about 10 mg to about 300 mg, two
times per day. ' .

The fused pyrrolocarbazole may be present in various
forms as will be appreciated by the skilled artisan. Such
forms include, but are not limited to, pharmaceutically
acceptable salts, prodrugs, polymorphs, stereoisomers, and
the like. As used herein, the term "pharmaceutically
acceptable" refers to those compounds, materials,
compositions, and/or dosage forms which are, within the
scope of sound medical judgment, suitable for contact with
the tissues of human beings and animals without excessive
toxicity, irritation, allergic response, or other problem
complications commensurate with a reasonable benefit/risk
ration.
As used herein, "pharmaceutically acceptable salts"
refer to derivatives of the disclosed compounds wherein
the parent compound is modified by making acid or base
salts thereof. Examples of pharmaceutically acceptable
salts include, but are not limited to, mineral or organic
acid salts of basic residues such as amines; alkali or
organic salts of acidic residues such as carboxylic acids;
and the like. The pharmaceutically acceptable salts
include the conventional non-toxic salts or the quaternary
ammonium salts of the parent compound formed, for example,
from non-toxic inorganic or organic acids. For example,
such conventional non-toxic salts include those derived
from inorganic acids such as hydrochloric, hydrobromic,
sulfuric, sulfamic, phosphoric, nitric and the like; and
the salts prepared from organic acids such as acetic,
propionic, succinic, glycolic, stearic, lactic, malic,
tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic,
phenylacetic, glutamic, benzoic, salicylic, sulfanilic, 2-
acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic,
ethane disulfonic, oxalic, isethionic, and the like.
The pharmaceutically acceptable salts of the present
invention can be synthesized from the parent compound
which contains a basic or acidic moiety by conventional

chemical methods. Generally, such salts can be prepared
by reacting the free acid or base forms of these compounds
with a stoichiometric amount of the appropriate base or
acid in water or in an organic solvent, or in a mixture of
the two. Generally, nonaqueous media like ether, ethyl
acetate, ethanol, isopropanol, or acetonitrile are
preferred. Lists of suitable salts are found in
Remington's Pharmaceutical Sciences, 17th ed., Mack
Publishing Company, Easton, PA, 1985, p. 1418, the
disclosure of which is hereby incorporated by reference.
The fused pyrrolocarbazoles of the present invention
may exist in prodrug form. As used herein, "prodrug" is
intended to include any covalently bonded carriers which
release the active parent drug according to Formula I or
other formulas or compounds of the present invention in
vivo when such prodrug is administered to a mammalian
subject. Since prodrugs are known to enhance numerous
desirable qualities of pharmaceuticals (e.g., solubility,
bioavailability, manufacturing, etc.) the compounds of the
present invention may be delivered in prodrug form". . Thus,
the present invention contemplates prodrugs of the claimed
compounds, compositions containing the same, and methods
of delivering the same. Prodrugs of a compound of the
present invention, for example Formula I, may be prepared
by modifying functional groups present in the compound in
such a way that the modifications are cleaved, either in
routine manipulation or in vivo, to the parent.compound.
Accordingly, prodrugs include, for example, compounds
of the present invention wherein a hydroxy, amino, or
carboxy group is bonded to any group that, when the
prodrug is administered to a mammalian subject, cleaves to
form a free hydroxyl, free amino, or carboxylic acid,
respectively. Examples include, but are not limited to,
acetate, formate and benzoate derivatives of alcohol and

amine functional groups; and alkyl, carbocyclic, aryl, and
alkylaryl esters such as methyl, ethyl, propyl, iso-
propyl, butyl, isobutyl, sec-butyl, tert-butyl,
cyclopropyl, phenyl, benzyl, and phenethyl esters, and the
like.
The compounds of the present invention may be
prepared in a number of ways well known to those skilled
•in the art. The compounds can be synthesized, for
example, by the methods described below, or variations
thereon as appreciated by the skilled artisan. All
processes disclosed in association with the present
invention are contemplated to be practiced on any scale,
including milligram, gram, multigram, kilogram,
multikilogram or commercial industrial scale.
It will be appreciated that the compounds of the
present invention may contain one or more asymmetrically
substituted carbon atoms, and may be isolated in optically
active or racemic forms. Thus, all chiral,
diastereomeric, racemic forms and all geometric isomeric
forms of a structure are intended, unless the specific
stereochemistry or isomeric form is specifically
indicated. It is well known in the art how to prepare and
isolate such optically active forms. For example,
mixtures of stereoisomers may be separated by standard
techniques including, but not limited to, resolution of
racemic forms, normal, reverse-phase, and chiral
chromatography, preferential salt formation,
recrystallization, and the like, or by chiral synthesis
either from chiral starting materials or by deliberate
synthesis of target chiral centers.
As will be readily understood, functional groups
present may contain protecting groups during the course of
synthesis. For example, the amino acid side chain
substituents of the compounds of Formula la can be
substituted with protecting groups such as

benzyloxycarbonyl or t-butoxycarbonyl groups. Protecting
groups are known per se as chemical functional groups that
can be selectively appended to and removed from
functionalities, such as hydroxyl groups and carboxyl
groups. These groups are present in a chemical compound
to render such functionality inert to chemical reaction
conditions to which the compound is exposed. Any of a
variety of protecting groups may be employed with the
present invention. Preferred protecting groups include
the benzyloxycarbonyl (Cbz; Z) group and the tert-
butyloxycarbonyl (Boc) group. Other preferred protecting
groups according to the invention may be found in Greene,
T.W. and Wuts, P.G.M., Protective Groups in Organic
Synthesis 2d. Ed., Wiley & Sons, 1991.
Fused pyrrolocarbazoles, such as indolocarbazoles may
be synthesized by methods taught, for example, in U.S.
Patent Nos. 4,923,986; 4,877,776; 5,093,330; 5,461,146;
5,468,872; 5,621,100; 5,621,101; 5,516,771; and 5,599,808;
and PCT publication Nos. WO 93/08809 and WO 97/46565, the
discloses of which are hereby incorporated herein by
reference in their entirety. Additional methods of
preparation are set forth in Moody et al., J. Org. Chem.
57:2105-2114 (1992), also incorporated herein by
reference.
Fused pyrrolocarbazoles, such as indenocarbazoles, as
well as additional compounds wherein Q is not a single
nitrogen, may be synthesized by methods taught, for
example, in U.S. Patent Nos. 5,475110; 5,591,855;
5,594,009; 5,705,511; 5,616,724; and 5,801,190; the
disclosures of which are hereby incorporated herein by
reference in their entirety.
Fused pyrrolocarbazoles, such as bridged
indenocarbazoles, may be prepared by methods taught, for
example, in U.S. Patent Application Serial No. 09/325,140,
v

the disclosure of which is hereby incorporated herein
by reference in its entirety.
The fused pyrrolocarbazoles disclosed in all foregoing
references are contemplated for use in the particle-forming
compositions of the present invention. Other exemplary
fused pyrrolocarbazoles are the ihdolocarbazoles set forth
in Tables I-A and I-B, wherein each entry corresponds to the
accompanying structure.






1IVa-l and IVa-4 are a 1.5 to 1.0 mixture of the components.
Certain preferred indolocarbazoles are compounds
designated by formula IIa-4, IIa-12, and IIa-51 having the .
following structures.

The non-aqueous particle-forming compositions of the
.invention provide greatly improved bioavailability of fused
pyrrolocarbazoles. For example, in water Compound IIa-12
has essentially _no solubility. In pure Tween 80, it has a
solubility of 28.43 mg/ml. When placed directly in an
aqueous solution of 2% Tween 80, Compound IIa-12 has a
solubility of 0.18 mg/ml. When initially dissolved in Tween

80 (a particle-forming composition) and subsequently diluted
50-fold with water (to produce an aqueous solution of 2%
Tween 80), the final solubility is 0.57 mg/ml.
Thus, dissolving the fused pyrrolocarbazole initially
in this particular particle-forming composition achieves a
better than 3.2-fold increase in fused pyrrolocarbazole
concentration in the final solution. Because initial
contact with an aqueous medium occurs in vitro or in vivo,
the non-aqueous particle-forming compositions are useful for
therapeutic administration of a fused pyrrolocarbazole to a
mammal.
The present invention also features a method of forming
a stable suspension of fused pyrrolocarbazole-containing
particles. The method typically includes: (a) dissolving an
fused pyrrolocarbazole in a non-aqueous liquid containing a
surfactant in an amount from about 20% to greater thaji 99%,
thereby forming an fused pyrrolocarbazole-containing,
particle-forming composition, and (b) contacting the
par tide-forming composition with an aqueous medium. The
particle-forming composition can be contacted with the
aqueous medium in vitro or in vivo. Preferably, the amount
of surfactant is at least 30%, and more preferably, it is at
least 40%, with 50% being particularly preferred. The
particle-forming composition optionally includes an organic
solvent, a lipid, an antioxidant, or a combination of those
components.
The present invention additionally features methods for
preparing the particle-forming compositions described
herein. In one embodiment, the first step involves heating
at least one organic solvent. Preferably the solvent is
heated at a temperature that either liquefies those solvents
that are solid or semi-solid at room temperature, or that
maximizes the solubilization of the active agent, but below
that which leads to degradation or instability of the
mixture. Some compositions may require addition of at least

one antioxidant to minimize degradation of the composition.
The second step involves mixing the active agent into the
heated solvent or heated solvent mixture. The active agent
includes the fused pyrrolocarbazoles disclosed herein.
Preferably the mixing is done at a high speed, with a high
shear mixing device or homogenizer to minimize the amount c
mixing time and to thoroughly disperse the active agent. •
Preferably the resultant mixture is homogenized mixture.
The third step involves addition' of the surfactant and any
additional excipients to the homogenized mixture.
Preferably the additional excipients are stirred into the
homogenized mixture, or mixed in at a low speed. The
resultant composition can be dispensed into gelatin or HPMC
capsules as a warm solution and allowed to solidify.
Alternatively, the solution can be slab cooled or spray
congealed and further processed by milling, granulating and
blending' and dry-powder filled into gelatin or HPMC
capsules.
In a further embodiment, the method for preparation of
the compositions of the present invention comprises heating
at least one organic solvent and.optionally one or more
antioxidants, followed by addition of a fused
pyrrolocarbazole and mixing with a high shear hodmogenizer,
followed by addition of one or more surfactants. In
particular, the present invention includes a method for
preparing a composition of comprising a fused
pyrrolocarbazole, at least one organic solvent, at least one
surfactant, and optionally at least one antioxidant wherein
the compassion is non-aqueous and particle-forming
comprising:
(a) heating the organic solvent and optionally
the antioxidant to about 50-90 °C to form a heated mixture;
(b) mixing the fused pyrrolocarbazole in the
heated mixture with a high shear homogenizer to form a
homogenized mixture; and

(c) mixing the surfactant to the homogenized
mixture.
Preferably, at least one antioxidant is heated with the
organic solvent. Preferably the temperature range is 50-70
°C, and more preferably is 60-70 °C.
The invention also features a method of treating a
neurological disease or disorder in a mammal, e.g., a human.
The method includes formulating a fused pyrrolocarbazole in
a non-aqueous, fused pyrrolocarbazole-containing, particle-
forming composition containing a surfactant in an amount of
at least 20% (w/w), and administering a therapeutically
effective amount of the particle-forming composition to the
mammal. In one embodiment of the method, the particle- ..
forming composition is contacted with an aqueous medium in
vitro, to form a stable suspension. A therapeutically
effective amount of he stable suspension is then
administered to the mammal. In an alternative embodiment, a
therapeutically effective amount of the particle-forming
composition is administered directly to the mammal, where, it
contacts an aqueous medium in vivo.
The invention also features a method of treating
cancer, including prostate cancer, in a mammal, e.g., a
human. The method includes formulating a fused
pyrrolocarbazole in a non-aqueous, fused pyrrolocarbazole-
containing, particle-forming composition containing a
surfactant in an amount of at least 20% (w/w) , and
administering a therapeutically effective amount of the
particle-forming composition to the mammal. In one
embodiment of the method, the particle-forming composition
is contacted with an aqueous medium in vitro, to form a
stable suspension. A therapeutically effective amount of
the stable suspension is then administered to the mammal.
In an alternative embodiment, a therapeutically effective
amount of the particle-forming composition is administered

directly to the mammal, where it contacts an aqueous medium
in vivo.
The invention is further illustrated by the following
examples. The examples are provided for illustration
purposes only, and they are not to be construed as limiting
the scope or content of the invention.
Examples
Formulations
Numerous particle-forming compositions according to the
invention have been formulated using various components in
• various amounts relative to one another. The formulations
for several particle-forming compositions are presented in
Table 2, below.






pyrrolocarbazole is compound IIa-4, IIa-12, or IIa-51. In
other embodiments, the polyethylene glycol has a molecular
weight from about 300 to 8000 Daltons, or preferably from
400-3350 Daltons, or more preferably from 400-1500 Daltons.
In other embodiments, the polyoxyl stearate is Myrj® 52.
In additional exemplary formulations, the fused
pyrrolocarbazole is compound IIa-12, and the polyethylene
glycol is PEG-400, PEG-600, PEG-1000, or PEG-1450 and the
surfactant is polyoxyl stearate is Myrj® 52. The ratio of.
polyethylene glycol:polyoxyl stearate ranges from 50:50 to
80:20, preferably the ratio is 50:50 or 80:20.
In other exemplary formulations, the organic solvent
can be a mixture of polyethylene glycols. In one
embodiment, the polyethylene glycols are selected from PEG-
400, PEG-600, PEG-1000 and PEG-1450. In other embodiments,
the mixture of polyethylene glycols can be PEG-400/PEG-1000,
PEG-400/PEG-1450, PEG-600/PEG-1000, PEG-600/PEG-1450. In
further embodiments, the surfactant is a polyoxyl stearate,
and. is preferably Myrj® 52. The ratio of a polyethylene
glycol mixture .-polyoxyl stearate ranges from 50:50 to 80:20,
preferably the ratio is 50:50 or 80:20. In certain
embodiments the composition comprises PEG-400:PEG-
1000 :polyoxyl stearate in a ratio of 25:25:50, or PEG-
400 : PEG-1450 :polyoxyl stearate in a ratio of 25:25:50, or
PEG-600:PEG-1000:polyoxyl stearate in a ratio of 25:25:50,
or PEG-600:PEG-1450:polyoxyl stearate in a ratio of
25:25:50. In other embodiments, the composition comprises
PEG-400: PEG-1000-.polyoxyl stearate in a ratio of 40:40:20,
or PEG-400:PEG-1450:polyoxyl stearate in a ratio of
40:40:20, or PEG-600:PEG-1000 :polyoxyl stearate in a ratio
of 40:40:20, or PEG-600:PEG-1450 :polyoxyl stearate in a
ratio of 40:40:20.

Bioavailability in Rats
Compound IIa-12 has displayed anticancer activity in a
rat tumor model of prostate cancer. A bioavailability study
involving Compound IIa-12 formulated in non-aqueous,
particle forming compositions was carried out using rats.
Pharmacokinetic parameters were measured following
intravenous (i.v.) administration and oral (p.o.)
administration to the rats. In addition, the absolute oral
bioavailability (F) of Compound IIa-12 was examined after
single administration of three different formulations.
Male Sprague-Dawley rats were given either a bolus i.v.
dose administered into the tail vein, or p.o. doses by
gavage in one of three formulations. The formulations were .
as described in Table 2A, below. Formulation (a) is not a
particle-forming composition.


Blood samples were collected at predetermined time
points. Plasma levels of Compound IIa-12 were determined by
HPLC. The mean values for the pharmacokinetic parameters
are summarized in Table 3, below. In Table 3, Cmax is peak
plasma concentration; Tmax is time to peak plasma
concentration; Tt/2 is apparent elimination half-life; AUCO-4
is area under the curve (serum cone. vs. time); and F% is
absolute bioavailability (total dose/AUC).



The oral bioavailability was lower, fox the .hydrophilic
formulation (a) than for the particle-forming compositions
(b) and (c). These data suggest that bioavailability in
rats may be improved significantly, i.e., more than 300%, b}
using the particle-forming composition of the invention.
Bioavailability in Dogs
Bioavailability of two liquid compound IIa-12
formulations was determined in fasted beagle dogs. Using a
crossover study design, two separate formulations,
formulations 16 and 18 from Table 1 were administered to a
group of six dogs. An intravenous dose was included as the
third formulation in the crossover design. Each dog
received a 10 mg dose of compound IIa-12. The plasma
concentrations, as determined by HPLC with fluorescence
detection, were normalized to a 1 mg/kg dose.
Pharmacokinetic parameters, including peak plasma
concentration (Craax) , time to peak plasma concentration
(Tmax) , apparent elimination half life (ti/2), area under the
curve (AUCO0) and the absolute bioavailability (F%) were
determined.
The plasma concentration time profile of compound TTa-
12 following a 1 mg/kg intravenous dose was fitted to a two
compartment open model with apparent volume of distribution
values of 0.96 L/kg (range 0.70-1.43 L/kg) and 1.35 L/kg
(range 0.69v-202 L/kg) for Vc and Vp, respectively. Plasma

concentrations declined with a terminal elimination half-
life of 1.16 hours (range 0.89-1.38 hours). The plasma
clearance of compound IIa-12 following a 1 mg/kg bolus
intravenous dose in dog was 0.79 L/hr«kg (range 0.40-1.08
L/hr.kg).
Compound IIa-12 was rapidly absorbed from both of the
liquid formulations with peak concentrations recorded within
two hours of dosing. Peak plasma concentrations averaged
269±175 ng/ml (range 94-469 ng/ml) and 219+143 n/ml (range
96-458 ng/ml) for the Tween/propylene carbonate/PG and
Tween/PG formulations, respectively. The absolute
bioavailability for the two formulations was very similar,
averaging 41.6±8.4 and 39.3±11.3 percent, respectively.
A separate bioavailability study involving Compound
IIa-12 formulated in non-aqueous, particle forming
compositions was carried out using dogs. Pharmacokinetic
parameters were measured following oral dosing of eight
separate formulations. Dosage was 50 mg of Compound IIa-12
per dog per day. Each formulation was evaluated in a group
of three dogs, using a parallel study design.
Beagle dogs weighing 7.9-14.2 kg were used in the
study. The dogs were fasted overnight prior to dosing, but
were permitted water ad libitum. Food was returned to the
animals at the completion of the study.
Each formulation (capsule or liquid) was administered
to a group of three dogs, in a dose of 50 mg. The
formulations were as described in Table 4, below.
Formulations C and D are not particle-forming compositions.




The reference formulation and Formulations A-C were
administered during the first week of the study.
Formulations D-G were administered following a one-week
wash-out period. The liquid formulations A, E, F, and G
.were thoroughly mixed prior to dosing. Each liquid dose
(2 ml) was placed in a syringe and squirted into the back of
the throat of each dog and followed by about 10 ml water. A
similar amount of water was administered to each dog
receiving a capsule formulation.
Sequential heparinized .blood samples were obtained from
the jugular vein of each dog prior to dosing and at 0.25,
:0.5, 1.0, 1,5, 2, 3, 4, 6, 9, and 15 hours after
administration of each formulation. The blood samples were
promptly chilled in an ice bath. Within two hours of
collection, the samples were centrifuged (2500 rpm for 10
min.), and the plasma was transferred to polypropylene tubes
and frozen at -20°C for later analysis.
Plasma levels of Compound IIa-12 were determined by
HPLC, as described below. The plasma concentration in each
sample was calculated by least squares linear regression
analysis (unweighted) of the peak area ratio
(parent/internal standard) of the spiked dog plasma

standards versus concentration. Plasma concentrations were
normalized to a 5 mg/kg dose.
The mean values (±SD, N=3) for the pharmacokinetic
parameters are summarized in Table 5, below. In Table 5,
Cmax is peak plasma concentration; Tmax is time to peak plasma
concentration; Tt/2 is apparent elimination half-life; AUC0-4
is area under the curve (plasma cone. vs. time); and
% Formulation H is relative bioavailability.



The area under the curve from 0 to t hours (last
measurable plasma concentration time point) after dosing was
calculated using the linear trapezoidal rule for the plasma- '
time profiles. The residual area extrapolated to infinity,
determined as the final measured plasma concentration (Ct)
divided by the terminal elimination rate constant, was added
to AUCO_t to produce the total area under the curve (AUCO-4) .
The bioavailability relative to that of formulation REF was
calculated by dividing the AUCO_4 of each formulation by that
obtained for formulation REF.
The plasma concentrations of Compound IIa-12 were
determined by reverse phase HPLC with fluorescence detection
following liquid-liguid extraction of the samples with ethyl
acetate-.hexane (1:1, by vol.) at neutral pH. Following
evaporation of the organic solvent, the samples were
reconstituted with sequential aliquots of methanol and 0.1%
trifluoroacetic acid in water (final ratio 1:1, by vol.).
The parent compound and internal standard (Compound IIa-4)
were separated from co-extracted plasma contaminants on a 25
cm x 4.6 cm 5 µm Prodigy column. The mobile phase consisted
of acetonitrile:methanol:0.01 tetramethylammonium
perchlorate in 0.1% aqueous trifluoroacetic acid (45:5:55).
Flow rate was 1.0 ml/min. Fluorescence detection (λem = 288
nm, Xex = 390 nm) was used for quantitation of the analytes.
This HPLC assay for indolocarbazoles was linear over the
concentration range of 0-5000 ng/ml. These data suggest
that bioavailability in dogs may be improved significantly,

i.e., by more than 300%, by using the particle-form
composition of the invention.
Bioavailability in Monkeys
A bioavailability study involving Compound IIa-51
ormulated in non-aqueous, particle forming compositions was
arried out using monkeys. Pharmacokinetic parameters were
...easured following intravenous (i.v.) administration and
oral {p.o.) administration. The test composition consisted
of four male cynomolgus monkeys. The i.v. dose was
administered as a bolus, at a dosage level of 3 mg/kg. The
oral doses were administered as soft gelatin capsules, at a
dosage level of 10 mg/kg (2 capsules/monkey/dose). The oral
formulations were as described in Table 6, below.


Blood samples were collected at predetermined time
oints. Plasma levels of Compound IIa-51 were determined by
everse phase HPLC. Pharmacokinetic parameters were
stimated by non-compartmental methods. The mean values for
he pharmacokinetic parameters are summarized in Table 7,
elow. In Table 7, Cmax is peak plasma concentration; Tmax is
ime to peak plasma concentration; Tt/2 is apparent
limination half-life; AUC is area under the curve; and P%
is absolute bioavailability.

Following an i.v. dose, the mean clearance value was
1.1 ±0.1 L/hr/kg, and the mean volume of distribution was
4.9 ±0.7 L/kg. The results demonstrate that after i.v.
administration. Compound IIa-51 had a large volume of
distribution, rapid clearance,, and moderate Tt/2 - Based on
oral dosing, formulation D had better bioavailability than
the other formulations examined in this experiment.
Compound IIa-51 was better absorbed from hydrophilic

particles than from lipophilic particles, indicating a
• formulation-dependent absorption.
As those skilled in the art will appreciate, numerous
modifications and variations of the present invention are
possible in light of the above teachings. It is therefore
understood that within the scope of the appended claims, the
invention may be practiced otherwise than as specifically
described herein, and the scope of the invention is intended
to encompass all such variations.

WE CLAIM;
1. A composition comprising a fused pyrrolocarbazole with the formula:

wherein R3 and R4 are selected from H, a straight chain, cyclic or
branched alkyl group having 1 to 8 carbon atoms, Cl, Br, CH2OH,
CH2SOCH2CH3, CH2SO2CH2CH3, NHCONHC6H5, CH2SCH2CH3,
CH2SC6H5, NHCO2CH3, CH2OC(=O)NHCH2CH3,N(CH3)2,CH=NNH,
CH2N(CH3)2, and CH2OCH2CH3; R7 is selected from H and alkyl; and
R15 and R16 are independently selected from H, alkyl, pH, CH2OH,
alkoxy, and CO2alkyl; or a stereoisomer or pharmaceutically
acceptable salt form thereof; at least 20% (w/w) of a poloxyl stearate;
and at least one polyethylene glycol.
2. The composition as claimed in claim 1 wherein the fused
pyrrolocarbazole is present at a concentration of 1 to 100 mg/mL.
3. The composition as claimed in claim 2 wherein the fusgd
pyrrolocarbazole is present at a concentration of 1 to 50 mg/mL.
4. The composition as claimed in claim 1 wherein the fused
pyrrolocarbazole has the formula:


5. The composition as claimed in claim 4 wherein the fused
pyrrolocarbazole has the formula:

6. The composition as claimed in claim 1 wherein the polyethylene
glycol has a molecular weight from 300 to 8000 Daltons.


7. The composition as claimed in claim 6 wherein the polyethylene
glycol has a molecular weight from 400 to 3350 Daltons.
8. The composition as claimed in claim 7 wherein the polyethylene
glycol has a molecular weight from 400 to 1500 Daltons.
9. The composition as claimed in claim 5 wherein the polyethylene
glycol is selected from PEG-400, PEG-600, PEG- 1000, and PEG-
1450.

10. The composition as claimed in claim 9 wherein the polyoxyl
stearate is polyoxyl 40 stearate.
11. The composition as claimed in claim 10 wherein the ratio of
polyethylene glycol: polyoxyl stearate ranges from 50:50 to 80:20.
12. The composition as claimed in claim 11 wherein the ratio of
polyethylene glycol: polyoxyl stearate is 50:50.
13. The composition as claimed in claim 11 wherein the ratio of
polyethylene glycol: polyoxyl stearate is 80:20.
14. The composition as claimed in claim 1 comprising a polyethylene
glycol mixture selected from PEG-400/PEG-1000, PEG-400/PEG-
1450, PEG-600/PEG-1000, and PEG-600/PEG-1450.
15. The composition as claimed in claim 14 wherein the ratio of the
polyethylene glycol mixture: polyoxyl stearate is from 50:50 to 80:20.

16. The composition as claimed in claim 15 wherein the ratio of the
polyethylene glycol mixture: polyoxyl stearate is 50:50.
17. The composition as claimed in claim 15 wherein the ratio of the
polyethylene glycol mixture: polyoxyl stearate is 80:20.
18. The composition as claimed in claim 16 wherein the composition
comprises PEG-400:PEG-1000: polyoxyl stearate in a ratio of
25:25:50.
19. The composition as claimed in claim 16 wherein the composition
comprises PEG-400:PEG-1450:polyoxyl stearate in a ratio of 25:25:50.
20. The composition as claimed in claim 17 wherein the composition
comprises PEG-400:PEG-1000:polyoxyl stearate in a ratio of 40:40:20.
21. The composition as claimed in claim 17 wherein the composition
comprises PEG-400:PEG-1450:polyoxyl stearate in a ratio of 40:40:20.


A composition comprising a fused pyrrolocarbazole with the formula:

wherein R3 and R4 are selected from H, a straight chain, cyclic or branched alkyl
group having 1 to 8 carbon atoms, CI, Br, CH2OH, CH2SOCH2CH3, CH2SO2CH2CH3
NHCONHC6H5, CH2SCH2CH3, CH2SC6H5, NHCO2CH3, CH2OC(=O)NHCH2CH3,
N(CH3)2, CH=NNH, CH2N(CH3)2 and CH2OCH2CH3; R7 is selected from H and
alkyl; and R15 and R16 are independently selected from H, alkyl, OH, CH2OH,
alkoxy, and CO2alkyl; or a stereoisomer or pharmaceutically acceptable salt form
thereof; at least 20% (w/w) of a poloxyl stearate; and at least one polyethylene
glycol.

Documents:

01583-kolnp-2006 abstract.pdf

01583-kolnp-2006 assignment.pdf

01583-kolnp-2006 claims.pdf

01583-kolnp-2006 correspondence others-1.1.pdf

01583-kolnp-2006 correspondence others.pdf

01583-kolnp-2006 description (complete).pdf

01583-kolnp-2006 form-1.pdf

01583-kolnp-2006 form-2.pdf

01583-kolnp-2006 form-3.pdf

01583-kolnp-2006 international search report.pdf

01583-kolnp-2006 international publication.pdf

01583-kolnp-2006 pct form.pdf

01583-kolnp-2006 priority document.pdf

01583-kolnp-2006- correspondence-1.2.pdf

01583-kolnp-2006-claims-1.1.pdf

01583-kolnp-2006-correspondence-1.3.pdf

01583-kolnp-2006-form-13.pdf

01583-kolnp-2006-form-18.pdf

1583-KOLNP-2006-ABSTRACT.pdf

1583-kolnp-2006-assignment 1.1.pdf

1583-KOLNP-2006-CANCELLED DOCUMENT.pdf

1583-KOLNP-2006-CLAIMS.pdf

1583-kolnp-2006-correspondence 1.1.pdf

1583-KOLNP-2006-CORRESPONDENCE.pdf

1583-KOLNP-2006-DESCRIPTION (COMPLETE).pdf

1583-kolnp-2006-examination report 1.1.pdf

1583-KOLNP-2006-FORM 1.pdf

1583-kolnp-2006-form 13 1.1.pdf

1583-kolnp-2006-form 18 1.1.pdf

1583-KOLNP-2006-FORM 2.pdf

1583-kolnp-2006-form 26 1.1.pdf

1583-kolnp-2006-form 3 1.1.pdf

1583-kolnp-2006-form 5 1.1.pdf

1583-kolnp-2006-granted-abstract.pdf

1583-kolnp-2006-granted-claims.pdf

1583-kolnp-2006-granted-description (complete).pdf

1583-kolnp-2006-granted-form 1.pdf

1583-kolnp-2006-granted-form 2.pdf

1583-kolnp-2006-granted-specification.pdf

1583-kolnp-2006-intenational publication 1.1.pdf

1583-kolnp-2006-international preliminary examination report 1.1.pdf

1583-kolnp-2006-international search report 1.1.pdf

1583-KOLNP-2006-OTHERS.pdf

1583-KOLNP-2006-PCT PRIORITY DOCUMENT NOTIFICATION.pdf

1583-kolnp-2006-reply to examination report 1.1.pdf

1583-KOLNP-2006-REPLY TO EXAMINATION REPORT.pdf

1583-kolnp-2006-translated copy of priority document 1.1.pdf


Patent Number 246980
Indian Patent Application Number 1583/KOLNP/2006
PG Journal Number 12/2011
Publication Date 25-Mar-2011
Grant Date 23-Mar-2011
Date of Filing 08-Jun-2006
Name of Patentee CEPHALON, INC.
Applicant Address 145 BRANDYWINE PARKWAY, WEST CHESTER, PA 19380
Inventors:
# Inventor's Name Inventor's Address
1 DAVE DICKASON 4739 VICBARB LANE, CINCINNATI, OHIO 45244
2 PIYUSH R. PATEL 716 SCOTT LANE, WALLINGFORD, PENNSYLVANIA 19806
3 BRADLEY T. MCINTYRE 3316 NORMA DRIVE, THORNDALE, PENNSYLVANIA 19372
PCT International Classification Number A61K 31/55
PCT International Application Number PCT/US2004/037928
PCT International Filing date 2004-11-12
PCT Conventions:
# PCT Application Number Date of Convention Priority Country
1 10/718,077 2003-11-20 U.S.A.