CCR5 RECEPTOR ANTAGONIST MOLECULES

MOLECULES ANTAGONISTES DU RECEPTEUR CCR5

English Abstract

Compounds and pharmaceutical compositions are provided for the treatment of
AIDS.

Claims

We Claim:

1. Compounds (including salts, prodrugs, etc.) as described in the above
disclosure.

2. Pharmaceutical compositions comprising a compound (including salts,
prodrugs,
etc.) as described in the above disclosure and a pharmaceutically acceptable
carrier.

3. The use of a compound (including salts, prodrugs, etc.) as described in the
above
disclosure for the treatement of AIDS.

53

Description

CA 02522632 2005-10-06
Title : CCR5 Receptor Antagonist Molecules
Background
HIV is a retrovirus which causes the disease commonly known as AIDS. There are
two
types of HIV: HIV-1 and HIV-2. HIV-1 is responsible for the vast majority of
AIDS in the
United States. HIV-2, seen more often in western Africa, and has a slower
course than
HIV-1.
Antiretroviral therapy has been effective in reducing morbidity and mortality
in HIV-1
infected patients. The emergence of resistance to existing drugs, combined
with the lack
of adherence due to complicated dosing regimens causing side effects, led to
an
increase in the number of patients who are currently failing therapy. The
elucidation of
the different mechanistic steps by which HIV enters the host cell revealed
many new
targets for pharmacological intervention. Inhibitors of these new targets most
likely will
not be cross resistant to protease as well as reverse transcriptase inhibitors
since they
will interfere at different steps in the life cycle of HIV. Three discrete
steps have been
recognized in the entry process of HIV. In the first step the attachment of
the HIV
envelop to the CD4 receptor on host cells that are susceptible to infection
takes place.
Attachment results in a conformational change in the HIV envelop that triggers
the
second step namely, interaction with a co receptor, CCRS or CXCR4. Finally,
fusion
takes place between gp4l; the viral envelop component, and the host cell
membrane
resulting in infection of the susceptible cell. Inhibitors of each of these
steps have been
identified and progressed into clinical trials. The most advanced has been the
T-20
peptide, a fusion inhibitor that has been approved by the FDA in early 2003
and
marketed as Fusion by Roche/Trimmers.
Another possible mechanism to combat this disease is through primary
prevention of the
infection through microbicides. Microbicides are compounds that can be applied
inside
the vagina or rectum to protect against sexually transmitted infections (STIs)
including
HIV. They can be formulated as gels, creams, films, or suppositories.
Microbicides may
or may not have spermicidal activity (contraceptive effect). At present, an
effective
microbicide is not available.
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CA 02522632 2005-10-06
The chemokine receptor, CCRS, is a member of the G protein-coupled receptor
superfamily. Chemokine receptors expressed on the surface of immune cells such
as
monocytes, macrophages, dendritic cells, and T cells are known to play a
critical role in
HIV infection and transmission. The prevalence of the CCR5432 homozygous
genotype,
amounting to about 1 % of the Caucasian population, and results in no major
impact on
health while being strongly protective against HIV infection made this target
very
attractive for pharmacological intervention. Furthermore, CCR5 knockouts in
mice have
a benign phenotype with a very subtle impact on immune function. Hence, an
antagonist
specific for CCR5 may cause few, if any, mechanism-based side effects. These
and
other studies established that HIV-1 virus invades macrophages and primary T-
cells by
binding to the cell surface protein CD4 and the chemokine receptor CCRS. Thus
small
molecule CCR5 receptor antagonists may inhibit HIV-1 viral invasion of CD4+
host
immune cells and provide a powerful new therapeutic avenue for HIV therapy
(Maeda K,
Nakata H, Ogata H, Koh Y, Miyakawa T, and Mitsuya H (2004) Current Opinion in
Pharmacology 4: 447-452)
The hallmark of inflammatory disease is the infiltration of inflammatory cells
into target
organs. CCR5 receptors have been reported to mediate cellular transfer in
inflammatory
diseases such as rheumatoid arthritis, arthritis, asthma, psoriasis, atopic
dermatitis, and
allergies. Thus inhibitors of CCR5 receptor are expected to be of therapeutic
benefit in
these diseases and in other disease conditions such as inflammatory bowel
syndrome,
multiple sclerosis, organ transplant rejection and graft versus (v.) host
disease.
The present invention relates to certain molecules as CCR5 receptor
antagonists,
pharmaceutical compositions containing the compounds, and methods of
treatments
using the compounds. The invention also relates to the use of a combination of
CCR5
receptor antagonists of this invention with one or more antiviral agents or
other agents in
the treatment of HIV-1. The invention further relates to the use of CCR5
receptor
antagonists of this invention, alone or in combination with other agents, in
the treatment
of rheumatoid arthritis, arthritis, atopic dermatitis, psoraiasis, asthma,
allergies, multiple
sclerosis, solid organ transplant rejection, and graft v. host disease. As
shall be
discussed herein below molecules of the present invention may, for example, be
derived
from plant cell culture(s).
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CA 02522632 2005-10-06
Summary of the Invention
Thus the present invention relates to molecules or compounds of general
formulae 17,
18, 19, 19a, 19b, 20, 21 and 22 as set forth below. The present invention
further relates
to the treatment of HIV comprising administering to a human in need of such
treatment
an effective dose of a CCR5 receptor antagonist as described herein (i.e.
compounds of
general formulae 17, 18, 19, 19a, 19b, 20, 21 and 22 and for example in
particular the
molecules or compounds as set forth in Table 1 below). In accordance with
another
aspect of the invention there is provided a pharmaceutical composition for
treatment of
HIV comprising a pharmaceutically acceptable carrier and (an effective amount
of) a
CCR5 receptor antagonist as described herein (i.e. compounds of general
formulae 17,
18, 19, 19a, 19b, 20, 21 and 22 and for example in particular the molecules or
compounds as set forth in Table 1 below). This invention also relates to the
use of
these compounds as microbicides i.e. in an appropriate pharmaceutical
composition.
In addition, another aspect of this invention relates to the use of the above
mentioned
compounds for the treatment of solid organ rejection, graft v. host disease,
arthritis,
rheumatoid arthritis, inflammatory bowel disease, atopic dermatitis,
psoriasis, asthma,
allergies, or multiple sclerosis, In other words to pharmaceutical composition
for
treatment of HIV comprising a pharmaceutically acceptable carrier and (an
effective
amount of) a CCR5 receptor antagonist as described herein for such
applications.
Detailed Description of the Invention
The compounds of the present invention may be used for treating HIV infection.
They
may preferably be administered in substantially pure form. By "substantially
pure" is
meant that a compound as described herein may either be synthesized or be
purified
from a naturally derived state such that the compound is associated with only
trace
quantities of undesired materials or compounds that occur with it as a result
of its
synthesis or derivation from a natural state (e.g., the undesired materials or
compounds
are present in amounts whereby no unacceptable effect on a patient may be
observed).
By "isolated" is meant that the compound is in a state which is free or
essentially free of
being associated with other undesired or unacceptable molecules or compounds.
If
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CA 02522632 2005-10-06
desired, synthesized, isolated or purified compounds may be present in a
formulation
along with a pharmaceutical carrier as described herein below.
Thus, for example, many of the compounds of the present invention may be
isolated or
extracted from natural sources such as, for example, Hernandia ovigera and
Oreocereus
bruennowii. Several suitable methods for extracting the compounds of the
present
invention are known in the art. A highly preferred extraction method typically
comprises
crushing of a desired plant material (e.g., plant cell culture), blending the
crushed
material with a polar, preferably aqueous, solvent, filtering the slurry of
crushed material
t0 and polar solvent and purifying the filtrate. Several purification methods
like one-step or
multi-step liquid-liquid extraction, solid phase extraction, chromatography
and the like
are known from the state of the art.
Compounds according to the invention may also be synthesized, for example
using
compounds as shown in table 1 below as starting intermediates. The compounds
according to the invention can also be semi-synthesized. Compounds of the
present
invention may for example be synthesized from intermediated compounds
extracted
from plant cell culture as described herein.
One of ordinary skill in the art will recognize that some of the compounds of
the present
invention may exist in different geometrical isomeric forms. In addition,
compounds of
the present invention may possess one or more asymmetric carbon atoms and may
thus
capable of existing in the form of optical isomers, as well as in the form of
racemic or
nonracemic mixtures thereof, and in the form of diastereomers and
diastereomeric
mixtures inter alia. All of these compounds, including cis isomers, traps
isomers,
diastereomic mixtures, racemates, nonracemic mixtures of enantiomers, and
substantially pure and pure enantiomers, are within the scope of the present
invention.
Substantially pure enantiomers for example are to be understood herein as
containing
no more than 5% w/w of the corresponding opposite enantiomer, preferably no
more
than 2%, most preferably no more than 1 %.
The optical isomers may be obtained by resolution of the racemic mixtures
according to
conventional processes, for example, by the formation of diastereoisomeric
salts using
an optically active acid or base or formation of covalent diastereomers.
Examples of
appropriate acids are tartaric, diacetyltartaric, dibenzoyltartaric,
ditoluoyltartaric and
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CA 02522632 2005-10-06
camphorsulfonic acid. Mixtures of diastereoisomers may be separated into their
individual diastereomers on the basis of their physical and/or chemical
differences by
methods known to those skilled in the art, for example, by chromatography or
fractional
crystallization. The optically active bases or acids may be then liberated
from the
separated diastereomeric salts. A different process for separation of optical
isomers
involves the use of chiral chromatography (e.g., chiral High Performance
Liquid
Chromatography (HPLC) columns), with or without conventional derivation,
optimally
chosen to maximize the separation of the enantiomers. Suitable chiral HPLC
columns
are manufactured by Diacel, e.g., Chiracel OD and Chiracel OJ among many
others, all
routinely selectable. Enzymatic separations, with or without derivitization,
may also
useful. The optically active compounds of the present invention may likewise
be
obtained by chiral syntheses utilizing optically active starting materials.
The present invention also relates to useful forms of the compounds as
disclosed herein,
such as pharmaceutically acceptable salts and prodrugs of all the compounds of
the
present invention. A prodrug is to be understood herein as a compound that the
body is
able to convert into active drug; stated in another way, a prodrug is a
pharmacological
substance (drug) which is administered in an inactive (or significantly less
active) form;
however, once administered, the prodrug is metabolised in the body (in vivo)
into an
active compound..
Pharmaceutically acceptable salts include those obtained by reacting the main
compound, functioning as a base, with an inorganic or organic acid to form a
salt, for
example, salts of hydrochloric acid, sulfuric acid, phosphoric acid, methane
sulfonic acid,
camphor sulfonic acid, oxalic acid, malefic acid, succinic acid and citric
acid.
Pharmaceutically acceptable salts also include those in which the main
compound
functions as an acid and is reacted with an appropriate base to form, e.g.,
sodium,
potassium, calcium, mangnesium, ammonium, and choline salts. Those skilled in
the art
will further recognize that additional salts of the claimed compounds may be
prepared by
reaction of the compounds with the appropriate inorganic or organic acid via
any of a
number of known methods. Alternatively, alkali and alkaline earth metal salts
are
prepared by reacting the compounds of the invention with the appropriate base
via a
variety of known methods.
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The following are further examples of acid salts that may be obtained by
reaction with
inorganic or organic acids: acetates, adipates, alginates, citrates,
aspartates, benzoates,
benzenesulfonates, bisulfates, butyrates, camphorates, digluconates,
cyclopentanepropionates, dodecylsulfates, ethanesulfonates, glucoheptanoates,
glycerophosphates, hemisulfates, heptanoates, hexanoates, fumarates,
hydrobromides,
hydroiodides, 2-hydroxy-ethanesulfonates, lactates, maleates,
methanesulfonates,
nicotinates, 2-naphthalenesulfonates, oxalates, palmoates, pectinates,
persulfates, 3-
phenylpropionates, picrates, pivalates, propionates, succinates, tartrates,
thiocyanates,
tosylates, mesylates and undecanoates.
Preferably, the salts formed may be pharmaceutically acceptable for
administration to
mammals. However, pharmaceutically unacceptable salts of the compounds may be
suitable as intermediates, for example, for isolating the compound as a salt
and then
converting the salt back to the free base compound by treatment with an
alkaline
reagent. The free base may then be, if desired, be converted to a
pharmaceutically
acceptable acid addition salt.
The compounds of the present invention may be used for treating HIV infection
alone or
with co- infections e.g., opportunistic infections such as viral, bacterial or
parasitic co-
infection. Thus another aspect of the invention relates to the administration
of the
compounds of the present invention along with an effective amount of an agent
to treat
the co-infection. Typical opportunistic infections ("OI's") and other
disorders commonly
present with HIV disease may be bacterial and mycobacterial infections such
as, for
example, Mycobacterium Avium Complex (MAC, MAI), Salmonellosis, Syphilis,
Neuroshyphilis, Turberculosis (TB) and Bacillary angiomatosis (cat scratch
disease);
fungal infections such as, for example, Aspergillosis, Candidiasis (thrush,
yeast
infection), Coccidioidomycosis, Cryptococcal Meningitis and Histoplasmosis;
malignancies such as, for example, Kaposi's Sarcoma, Lymphoma, Systemic Non-
Hodgkin's Lymphoma (NHL) and Primary CNS Lymphoma; protozoal infections such
as, for example, Cryptosporidiosis, Isosporiasis, Microsporidiosis,
Pneumocystis Carinii
Pneumonia (PCP) and Toxoplasmosis; viral infections such as, for example,
Cytomegalovirus (CMV), Hepatitis, Herpes Simplex (HSV, genital herpes), Herpes
Zoster (HZV, shingles), Human Papiloma Virus (HPV, genital warts, cervical
cancer),
Molluscum Contagiosum, Oral Hairy Leukoplakia (OHL) and Progressive Multifocal
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Leukoencephalopathy (PML); neurological conditions such as, for example, AIDS
Dementia Complex (ADC) and Peripheral Neuropathy. Other conditions and
complications of HIV infection include, for example, Apthous Ulcers,
malabsorption,
depression, diarrhea, thrombocytopenia, wasting syndrome, idiopathic
thrombocytopenic
purpura, Listeriosis, pelvic inflammatory disease, Burkitt's lymphoma and
immunoblastic
lymphoma. Treatment for these opportunistic infections and other disorders are
conventionally known. Suitable agents, which may be co-administered or used in
combination therapy with the compounds of the present invention, can be found,
for
example, at http://www.aegis.com/topics/oi/.
The activity of the compounds of the present invention may be determined by
any one of
a number of known assays available to one skilled in the art. The activity of
the
compounds according to the invention may also be determined by assays such as
those
described in the Examples.
The compounds (including salts, prodrugs, etc.) of the invention may be
administered
alone, but preferably as an active ingredient of a formulation. Thus, the
present
invention also includes pharmaceutical compositions containing a compound of
the
present invention and one or more pharmaceutically acceptable carriers,
excipients etc.
Numerous standard references are available that describe procedures for
preparing
various formulations suitable for administering the compounds according to the
invention. Examples of potential formulations and preparations are contained,
for
example, in the Handbook of Pharmaceutical Excipients, American Pharmaceutical
Association (current edition); Pharmaceutical Dosage Forms: Tablets
(Lieberman,
Lachman and Schwartz, editors) current edition, published by Marcel Dekker,
Inc., as
well as Remington's Pharmaceutical Sciences (Arthur Osol, editor), 1553-1593
(current
edition).
In view of their potentially high degree of efficacy in reducing HIV viral
load, the
compounds of the present invention may be administered to anyone requiring or
desiring
reduction in HIV viral load, including patients having HIV with co-infection.
Administration
may be accomplished according to patient needs, for example, orally, nasally,
parenterally (subcutaneously, intravenously, intramuscularly, intrasternally
and by
infusion), by inhalation, rectally, vaginally, topically, locally,
transdermally, and by ocular
administration.
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Various solid oral dosage forms can be used for administering compounds of the
invention including such solid forms as tablets, gelcaps, capsules, caplets,
granules,
lozenges and bulk powders. The compounds (including salts, prodrugs, etc.) of
the
present invention can be administered alone or combined with various
pharmaceutically
acceptable carriers, diluents (such as sucrose, mannitol, lactose, starches)
and
excipients known in the art, including but not limited to suspending agents,
solubilizers,
buffering agents, binders, disintegrants, preservatives, colorants,
flavorants, lubricants
and the like. Time release capsules, tablets and gels are also advantageous in
administering the compounds of the present invention.
Various liquid oral dosage forms may also be used for administering compounds
of the
invention, including aqueous and non-aqueous solutions, emulsions,
suspensions,
syrups, and elixirs. Such dosage forms may also contain suitable inert
diluents known in
the art such as water and suitable excipients known in the art such as
preservatives,
wetting agents, sweeteners, flavorants, as well as agents for emulsifying
and/or
suspending the compounds of the invention. The compounds of the present
invention
may be injected, for example, intravenously, in the form of an isotonic
sterile solution.
Other preparations are also possible.
Suppositories for rectal administration of the compounds of the present
invention may be
prepared by mixing the compound with a suitable excipient such as cocoa
butter,
salicylates and polyethylene glycols. Formulations for vaginal administration
can be in
the form of a pessary, tampon, cream, gel, paste, foam, or spray formula
containing, in
addition to the active ingredient, such suitable carriers as are known in the
art.
For topical administration the pharmaceutical composition may be in the form
of creams,
ointments, liniments, lotions, emulsions, suspensions, gels, solutions,
pastes, powders,
sprays, and drops suitable for administration to the skin, eye, ear or nose.
Topical
administration may also involve transdermal administration via means such as
transdermal patches.
Aerosol formulations suitable for administering via inhalation also may be
made. For
example, the compounds according to the invention can be administered by
inhalation in
the form of a powder (e.g., micronized) or in the form of atomized solutions
or
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CA 02522632 2005-10-06
suspensions. The aerosol formulation may be placed into a pressurized
acceptable
propellant.
The compounds (including salts, prodrugs, etc.) of the present invention may
be
administered as the sole active agent or in combination with one or more,
preferably one
to eight, anti-viral agents useful in anti-HIV-1 therapy e.g., protease
inhibitors, reverse
transcriptase inhibitors, fusion inhibitors ("FI"s) or other antiviral drugs
such as ribavirin.
Suitable antiviral agents which may be co-administered or used in combination
therapy
include, for example, suitable nucleoside and nucleotide reverse transcriptase
inhibitors
("NRTI" s) including, for example, zidovudine (AZT) available under the
RETROVIR
tradename from Glaxo-Welicome Inc., Research Triangle, N.C. 27709; didanosine
(ddl)
available under the VIDEX tradename from Bristol-Myers Squibb Co., Princeton,
N.J.
08543; zalcitabine (ddC) available under the HIVID tradename from Roche
Pharmaceuticals, Nutley, N.J. 07110; stavudine (d4T) available under the ZERIT
trademark from Bristol-Myers Squibb Co., Princeton, N.J. 08543; lamivudine
(3TC)
available under the EPIVIR tradename from Glaxo-Wellcome Research Triangle,
N.C.
27709; abacavir (1592U89) disclosed in W096/30025 and available under the
ZIAGEN
trademark from Glaxo-Wellcome Research Triangle, N.C. 27709; adefovir
dipivoxil
[bis(POM)-PMEA] available under the PREVON tradename from Gilead Sciences,
Foster City, Calif. 94404; lobucavir (BMS-180194), a nucleoside reverse
transcriptase
inhibitor disclosed in EP-0358154 and EP-0736533 and under development by
Bristol-
Myers Squibb, Princeton, N.J. 08543; BCH-10652, a reverse transcriptase
inhibitor (in
the form of a racemic mixture of BCH-10618 and BCH-10619) under development by
Biochem Pharma, Laval, Quebec H7 V 4A7, Canada; emitricitabine [(-)-FTC]
licensed
from Emory University under Emory Univ. U.S. Pat. No. 5,814,639 and under
development by Triangle Pharmaceuticals, Durham, N.C. 27707; beta-L-FD4 (also
called beta-L-D4C and named beta-L-2',3'-dicleoxy-5-fluoro-cytidene) licensed
by Yale
University to Vion Pharmaceuticals, New Haven Conn. 06511; DAPD, the purine
nucleoside, (-)-beta-D-2,6,-diamino-purine dioxolane disclosed in EP 0656778
and
licensed by Emory University and the University of Georgia to Triangle
Pharmaceuticals,
Durham, N.C. 27707; and lodenosine (FddA), 9-(2,3-dideoxy-2-fluoro-b-D-threo-
pentofuranosyl)adenine, an acid stable purine-based reverse transcriptase
inhibitor
discovered by the NIH and under development by U.S. Bioscience Inc., West
Conshohoken, Pa. 19428.
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Typical suitable non-nucleoside reverse transcriptase inhibitors ("NNRTI"s)
include
nevirapine (BI-RG-587) available under the VIRAMUNE tradename from Boehringer
Ingelheim, the manufacturer for Roxane Laboratories, Columbus, Ohio 43216;
delaviradine (BHAP, U-90152) available under the RESCRIPTOR tradename from
Pharmacia & Upjohn Co., Bridgewater N.J. 08807; efavirenz (DMP-266) a
benzoxazin-2-
one disclosed in W094/03440 and available under the SUSTIVA tradename from
DuPont Pharmaceutical Co., Wilmington, Del. 19880-0723; PNU-142721, a
furopyridine-
thio-pyrimide under development by Pharmacia and Upjohn, Bridgewater N.J.
08807;
AG-1549 (formerly Shionogi #S-1153); 5-(3,5-dichlorophenyl)-thio-4-isopropyl-1-
(4-
t0 pyridyl)methyl-1 H-imidazol-2-ylmethyl carbonate disclosed in WO 96/10019
and under
clinical development by Agouron Pharmaceuticals, Inc., LaJolla Calif. 92037-
1020; MKC-
442 (1-(ethoxy-methyl)-5-(1-methylethyl)-6-(phenylmethyl)-(2,4(iH,3H)-
pyrimidinedione)
discovered by Mitsubishi Chemical Co. and under development by Triangle
Pharmaceuticals, Durham, N.C. 27707; and (+)-calanolide A (NSC-675451 ) and B,
coumarin derivatives disclosed in NIH U.S. Pat. No. 5,489,697 licensed to Med
Chem
Research, which is co-developing (+) calanolide A with Vita-Invest as an
orally
administrable product.
Protease inhibitors ("PI") include compounds having a peptidomimetic
structure, high
molecular weight (7600 daltons) and substantial peptide character, e.g.
CRIXIVAN
(available from Merck) as well as nonpeptide protease inhibitors e.g.,
VIRACEPT
(available from Agouron).
Typical suitable Pls include, for example, saquinavir (Ro 31-8959) available
in hard gel
capsules under the INVIRASE tradename and as soft gel capsules under the
FORTOVASE tradename from Roche Pharmaceuticals, Nutley, N.J. 07110-1199;
ritonavir (ABT-538) available under the NORVIR tradename from Abbott
Laboratories,
Abbott Park, III. 60064; indinavir (MK-639) available under the CRIXIVAN
tradename
from Merck & Co., Inc., West Point, Pa. 19486-0004; nelfnavir (AG-1343)
available
under the VIRACEPT tradename from Agouron Pharmaceuticals, Inc., LaJolla
Calif.
92037-1020; amprenavir (141 W94), tradename AGENERASE, a non-peptide protease
inhibitor under development by Vertex Pharmaceuticals, Inc., Cambridge, Mass.
02139-
4211 and available from Glaxo-Wellcome, Research Triangle, N.C. under an
expanded
access program; lasinavir (BMS-234475) available from Bristol-Myers Squibb,
Princeton,
N.J. 08543 (originally discovered by Novartis, Basel, Switzerland (CGP-61755);
DMP-

CA 02522632 2005-10-06
450, a cyclic urea discovered by Dupont and under development by Triangle
Pharmaceuticals; BMS-2322623, an azapeptide under development by Bristol-Myers
Squibb, Princeton, N.J. 08543, as a 2nd-generation HIV-1 PI; ABT-378 under
development by Abbott, Abbott Park, III. 60064; and AG-1549 an orally active
imidazole
carbamate discovered by Shionogi (Shionogi #S-1153) and under development by
Agouron Pharmaceuticals, Inc., LaJolla Calif. 92037-1020.
Other antiviral agents include, for example, hydroxyurea, ribavirin, IL-2, IL-
12,
pentafuside and Yissum Project No.11607. Hydroyurea (Droxia), a ribonucleoside
triphosphate reductase inhibitor, the enzyme involved in the activation of T-
cells, was
discovered at the NCI and is under development by Bristol-Myers Squibb; in
preclinical
studies, it was shown to have a synergistic effect on the activity of
didanosine and has
been studied with stavudine. IL-2 is disclosed in Ajinomoto EP-0142268, Takeda
EP-
0176299, and Chiron U.S. Pat. Nos. RE 33,653, 4,530,787, 4,569,790, 4,604,377,
4,748,234, 4,752,585, and 4,949,314, and is available under the PROLEUKIN
(aldesleukin) tradename from Chiron Corp., Emeryville, Calif. 94608-2997 as a
lyophilized powder for IV infusion or sc administration upon reconstitution
and dilution
with water; a dose of about 1 to about 20 million IU/day, sc is preferred; a
dose of about
15 million IU/day, sc is more preferred. IL-12 is disclosed in W096/25171 and
is
available from Roche Pharmaceuticals, Nutley, N.J. 07110-1199 and American
Home
Products, Madison, N.J. 07940; a dose of about 0.5 microgram/kg/day to about
10
microgram/kg/day, sc is preferred. Pentafuside (DP-178, T-20) a 36-amino acid
synthetic
peptide, disclosed in U.S. Pat. No. 5,464,933 licensed from Duke University to
Trimeris
which is developing pentafuside in collaboration with Duke University;
pentafuside acts
by inhibiting fusion of HIV-1 to target membranes. Pentafuside (3-100 mg/day)
is given
as a continuous sc infusion or injection together with efavirenz and 2 PI's to
HIV-1
positive patients refractory to a triple combination therapy; use of 100
mg/day is
preferred. Yissum Project No. 11607, a synthetic protein based on the HIV-1
Vif protein,
is under preclinical development by Yissum Research Development Co., Jerusalem
91042, Israel. Ribavirin, 1-f3-D-ribofuranosyl-1H-1,2,4-triazole-3-
carboxamide, is
available from ICN Pharmaceuticals, Inc., Costa Mesa, Calif; its manufacture
and
formulation are described in U.S. Pat. No. 4,211,771.
The term "anti-HIV-1 therapy" as used herein means any anti-HIV-1 drug found
useful
for treating HIV-1 infections in human beings either alone, or as part of
multidrug
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combination therapies, especially the HAART (highly active antiretroviral
therapy) triple
and quadruple combination therapies; HAART - a combination of protease
inhibitors
taken with reverse transcriptase inhibitors. Typical suitable known anti-HIV-1
therapies
include, but are not limited to multidrug combination therapies such as (i) at
least three
anti-HIV-1 drugs selected from two NRTIs (non-nucleoside reverse transcriptase
inhibitor - an antiviral drug used against HIV; binds directly to reverse
transcriptase and
prevents RNA conversion to DNA), one PI, a second PI, and one NNRTI (an
antiviral
drug used against HIV; is incorporated into the DNA of the virus and stops the
building
process; results in incomplete DNA that cannot create a new virus); and (ii)
at least two
anti-HIV-1 drugs selected from NNRTIs and Pls. Typical suitable HAART-
multidrug
combination therapies include:
~ (a) Triple combination therapies such as two NRTIs and one PI; or (b) two
NRTIs
and one NNRTI; and (c) quadruple combination therapies such as two NRTIs,
one PI and a second PI or one NNRTI. In treatment of naive patients, it is
preferred to start anti-HIV-1 treatment with the triple combination therapy;
the use
of two NRTIs and one PI is prefered unless there is intolerance to Pls. Drug
compliance is essential. The CD4+ and HIV-1-RNA plasma levels should be
monitored every 3-6 months. Should viral load plateau, a fourth drug, e.g.,
one PI
or one NNRTI could be added. Typical therapy schemes are described below:
ANTI-HIV-1 MULTI DRUG COMBINATION THERAPY SCHEMES
A. Triple Combination Therapies
~ 1. Two NRTIs +one PI
~ 2. Two NRTIs +one NNRTI
B. Quadruple Combination Therapies
~ Two NRTIs+one PI+a second PI or one NNRTI
C. ALTERNATIVES:
~ Two NRTI
~ One NRTI +one PI
~ Two PI's~one NRTI or NNRTI
~ One PI +one NRTI +one NNRTI
12

CA 02522632 2005-10-06
~ One PI +one NRTI +one NNRTI + one FI
In such combinations, each active ingredient can be administered either in
accordance
with their usual dosage range or a dose below its usual dosage range.
The dosages of the compounds of the present invention depend upon a variety of
factors including the particular syndrome to be treated, the severity of the
symptoms, the
route of administration, the frequency of the dosage interval, the particular
compound
utilized, the efficacy, toxicology profile, pharmacokinetic profile of the
compound, and the
presence of any deleterious side-effects, among other considerations.
The compounds of the invention may typically be administered at
therapeutically
effective dosage levels and in a mammal an amount customary for HIV viral load
reduction such as those known compounds mentioned above. For example, the
compounds may be administered, in single or multiple doses, by oral
administration at a
dosage level of, for example, 0.01-100 mg/kg/day, such as 0.1-70 mg/kg/day,
and in
particular 0.5-10 mg/kg/day. Unit dosage forms may contain, for example, 0.1-
50 mg of
active compound. For intravenous administration, the compounds may be
administered,
in single or multiple dosages, at a dosage level of, for example, 0.001-50
mg/kg/day,
such as 0.001-10 mg/kg/day, and in particular 0.01-1 mg/kg/day. Unit dosage
forms
may contain, for example, 0.1-10 mg of active compound. A therapeutically
effective
amount may be an amount sufficient to lower HIV-1 RNA plasma levels.
In carrying out the procedures for the preparation of compounds of the present
invention
it is of course to be understood that reference to particular buffers, media,
reagents,
cells, culture conditions and the like are not intended to be limiting, but
are to be read so
as to include all related materials that one of ordinary skill in the art
would recognize as
being of interest or value in the particular context in which that discussion
is presented.
For example, it is often possible to substitute one buffer system or culture
medium for
another and still achieve similar, if not identical, results. Those of skill
in the art will have
sufficient knowledge of such systems and methodologies so as to be able,
without
undue experimentation, to make such substitutions as will optimally serve
their purposes
in using the methods and procedures disclosed herein.
13

CA 02522632 2005-10-06
The present invention will now be further described by way of the following
non-limiting
examples. In applying the disclosure of these examples, it should be kept
clearly in mind
that other and different embodiments of the methods disclosed according to the
present
invention will no doubt suggest themselves to those of skill in the relevant
art.
In the foregoing and in the following examples, all temperatures are set forth
uncorrected
in degrees Celsius; and, unless otherwise indicated, all parts and percentages
are by
weight.
It is to further be understood herein, that if a "class", "range", "group of
substances", etc.
is mentioned with respect to a particular characteristic (e.g., temperature,
concentration,
time and the like) of the present invention, the present invention relates to
and explicitly
incorporates herein each and every specific member and combination of sub-
classes,
sub-ranges or sub-groups therein whatsoever. Thus, any specified class, range
or
group is to be understood as a shorthand way of referring to each and every
member of
a class, range or group individually as well as each and every possible sub-
class, sub-
range or sub-group encompassed therein; and similarly with respect to any sub-
class,
sub-ranges or sub-groups therein. Thus, for example,
with respect to the number of carbon atoms, the mention of the range of 1 to 6
carbon atoms is to be understood herein as incorporating each and every
individual number of carbon atoms as well as sub-ranges such as, for example,
1
carbon atoms, 3 carbon atoms, 4 to 6 carbon atoms, etc.;
with respect to reaction or extraction time, a time of 1 minute or more is to
be
understood as specifically incorporating herein each and every individual
time, as
well as sub-range, above 1 minute, such as for example 1 minute, 3 to 15
minutes, 1 minute to 20 hours, 1 to 3 hours, 16 hours, 3 hours to 20 hours
etc.;
etc...
It is in particular to be understood herein that for any group or range, no
matter how
defined, a reference thereto is a shorthand way of mentioning and including
herein
each and every individual member described thereby as well as each and every
possible
class or sub-group or sub-class of members whether such class or sub-class is
defined
as positively including particular members, as excluding particular members or
a
14

CA 02522632 2005-10-06
combination thereof; for example an exclusionary definition for a formula may
read as
follows: "provided that when one of A and B is -X and the other is Y, - X may
not be Z ".
The entire disclosures of all applications, patents and publications, cited
above and
below, are hereby incorporated by reference.
EXAMPLE I
Cell line generation
General description
Biological material is obtained from commercial seed or plant sources, the
Jardin de
Botanique de Montreal or naturally collected. All culture protocols are based
on
methods traditionally used for the culture of plant cells. See for example
R.A. Dixon Ed:
Plant Cell Culture a practical approach.1995 IRL Press Limited, Oxford.
Thus generally speaking, biological materials (plant material, seeds, etc.)
may be
sterilized with 5% sodium hypochlorite containing 1 % surfactant (e.g. Triton
X-100).
Sterilized materials are placed on typical media solidified with agar (0.8%)
or phytagel
(0.4%). Medium may consist of the inorganic salts proposed by Gamborg et al.
(1968)
Nutrient requirements of suspension cultures of soybean root cells. Exp. Cell
Res. 50:
151-158 or Murashige and Skoog (1962) A revised medium for rapid growth and
bioassays with tobacco tissue cultures. Physiol. Plant. 15: 473-497, 3%
sucrose and
various combinations of plant growth regulators (including 2,4-diphenoxyacetic
acid,
naphthaleneacetic acid, benzyladenine, kinetin, etc.) to select for suitable
cell lines.
Cell lines (and medium) may be selected from these plates based solely on the
growth
of the resulting calli, generated from the sterilized material, and may be
subsequently
transferred (every two months) onto the selected medium. Once the cell lines
are
stabilized (6-8 transfers) they are initiated into liquid cultures using the
identical medium
as for the solid transfer without the solidifying agent. Once fully
established (3 months or
more) cell lines are then scaled up by serial transfer (every fourteen days)
into larger

CA 02522632 2005-10-06
Erlenmeyer flasks. Cell lines are prepared for biosynthesis of metabolites by
culturing
into 1 L flasks for use as inocula.
More particularly,
a) for Hernandia ovigera
1-lernandia ovigera seeds (Banana Tree Inc.) were washed (2 min) with 70%
EtOH,
surface sterilized with 5% sodium hypochlorite containing 1 % Triton X-100 (60
min) and
washed three times with sterile water. Sterile seeds were placed on wetted
filter paper
(Whatman #4) in Petri dishes, sealed with parafilm and placed at room
temperature until
germination. Seedlings' meristems were cut with a sterile scalpel and place on
medium
solidified with 0.8% phytagel (Sigma). The medium consisted of the inorganic
salts
proposed by Gamborg et al. (1968) [Nutrient requirements of suspension
cultures of
soybean root cells. Exp. Cell Res. 50: 151-158], 3% sucrose, 0.2 mg/L of
dichlorophenoxyacetic acid and 0.1 mg/L kinetin. Callus developed on this
medium
were subsequently transferred unto the identical medium every two months. Once
the
cell lines were stabilized (6-8 transfers) they were initiated into liquid
cultures using the
identical medium as above without the solidifying agent. These cell lines are
maintained
by culture every 2 weeks by transfer of material (30%) into fresh medium. Once
fully
established (3 months) cell lines are then scaled up by serial transfer (every
fourteen
days) into larger Erlenmeyer flasks at the identical 30% inocula to medium
ratio. Finally,
cell lines are prepared for biosynthesis of metabolites of interest by
culturing into 1 L
flasks, as above, for use as inocula.
b) for Oreocereus bruennowii
Oreocereus bruennowii plants were purchased (Rona Inc.). Leaf material was
washed
(2 min) with 70% EtOH, surface sterilized (30 min) with 5% sodium hypochlorite
containing 1 % Triton X-100 and washed repeatedly (3x) with sterile water.
Plant
material was cut with a sterile scalpel and placed on medium solidified with
0.8%
phytagel. Medium consisted The medium consisted of the inorganic salts
proposed by
Gamborg et al. (1968) [Nutrient requirements of suspension cultures of soybean
root
16

CA 02522632 2005-10-06
cells. Exp. Cell Res. 50: 151-158], 3% sucrose, 1.0 mg/L of
dichlorophenoxyacetic acid
and 0.1 mg/L kinetin. Established cell cultures were treated identically as
above.
Example II
Biosynthesis
Compound biosynthesis may be conducted in 4 L sterile glass bioreactors
equipped with
an impeller, dissolved oxygen probe, inoculating and sampling ports. These
bioreactors
may be connected to peristaltic pumps for feeding nutrients (inorganic salts,
glucose,
etc), two mass flow controllers supply and control the dissolved oxygen and
carbon
dioxide concentrations at 120% and 4%, respectively. Finally the gas exhaust
may be
connected to a carbon dioxide monitor. All instrumentation may be connected to
a real
time control system to perform control algorithms and data monitoring.
All components of the bioreactor may be either glass or stainless steel and
are steam
sterilized. Cultures were inoculated with a suitable volume of a plant cell
suspension (as
mentioned above) from 1 L flasks (30%) into defined medium to obtain levels of
the
inorganic salts proposed by Gamborg et al. (1968) [Nutrient requirements of
suspension
cultures of soybean root cells Exp. Cell Res. 50: 151-158], 3% glucose, and
0.2 mg/L of
dichlorophenoxyacetic acid and 0.1 mg/L kinetin. Sampling 0100 mL) is done
every 48
h for measurement of biomass concentrations (wet weight and dry weight) and
nutrient
level determinations (HPLC, Dionex). All off-line and on-line information is
collected by
an expert control system allowing quality control of the bioprocess. This
system also
adjusts rate and composition of nutrient feeds to optimize levels present at
inoculation.
Once optimal growth is achieved secondary metabolites are induced by
elicitation of
secondary metabolism. Cells may be elicited with chitin (0.9% v/v), the
addition of a
concentrated nutrient solution, and absorbent resin (100 g XAD-7, 100 g XAD-
16: Rhom
& Hass)). The concentrated nutrient solution consisted of inorganic salts
based on
Murashige and Skoog (1962) A revised medium for rapid growth and bioassays
with
tobacco tissue cultures. Physiol. Plant. 15: 473-497. Concentrations were
adjusted to
achieve nutrient levels typical of inoculation, however, glucose levels were
adjusted to
5%. All solutions during the production phase were and plant growth regulator
and
phosphate free.
17

CA 02522632 2005-10-06
After 10 days of production, the culture is harvested by filtration to
separate the solid
phase (cells and resin), which contains the compounds of interest, from the
medium.
This material is used for the isolation of active compounds.
Example III
Biochemical Assay
A High-Throughput screen for inhibitors of ['251]MIP-1a binding to Chinese
hamster ovary
(CHO) cell membranes expressing the chemokine CCR5 receptor were performed as
described by Strizki et al. (Proc. Natl. Acad. Sci USA 98: 12717-127723).
Briefly, 1 p,g
CHO cell membranes over expressing the CCR5 receptor was incubated with 10 p,g
of
plant cell fractions in 100 p1 reaction mixture containing 50 mM HEPES pH 7.4,
5 mM
MgCl2, 1 mM CaCl2, 0.2% Bovine serum albumin, 2% Dimethylsulfoxide, and about
50
pM ['251]MIP-1a (PerkiElmer). Reactions were carried out for 90 min at room
temperature and were terminated by rapid filtration over UnifiIterR-96 GF/C
filter plates
(PerkinElmer) pre-soaked in 0.3% Polyethyleneimine and washed four times with
1 ml
cold (4 °C) buffer containing 10 mM HEPES pH 7.4, 500 mM NaCI, and 0.1
% Bovine
serum albumin. Fifty pL MicroscintT"" 0 (PerkinElmer) was added to dried
filter plate
wells and radioactivity retained on filters was counted in TopCount NXT HTS
Microplate
Scintillation and Luminescence counter (PerkinElmer). Non-specific binding was
determined in the presence of 1 p,M SCH-351125. About 110,000 fractions
derived from
plant cell cultures present in the library were screened and several fractions
were
obtained that show greater than 50% inhibition of ['251]MIP-1a binding to the
CCR5
receptor. These fractions were further purified and the active compounds were
identified.
Finally the structures of these compounds were elucidated.
Example IV
Chemistry
General Protocol for Isolation of Active compounds from Plant cell cultures
Compounds 1-8 (see table 1 below) were isolated from the extract of Hernandia
ovigera
plant cell cultures. The active fractions identified by HTS were further
purified using
preparative HPLC (C-18 Luna, ACE and/or Synergy columns) with
water/acetonitrile
18

CA 02522632 2005-10-06
gradients. Active fractions identified from Oreocereus bruennowii were treated
in the
same way as Hernandia ovigera to isolate compounds 10-14 (see table 1 below).
Compound 9 (Totarol) is a commercially available plant natural product. It was
first
isolated from the bark of Podocarpus nagi (Podocarpaceae) as an antimicrobial
agent
(Kubo I, Muroi H, and Himejima M (1992) J. Nat. Prod. 55, 1436-40). Up until
the
present invention, no CCR5 activity has been described for compound 9.
Thus more particularly, the solid materials harvested from biosynthesis were
all treated
in an identical manner. The solid material was extracted five times using
acidified
methanol (0.1 % v/v acetic acid)). The resulting methanolic extracts are
combined,
concentrated in vacuo and dried onto C4 stationary phase material (Macherey &
Nagel
Polygoprep RP4 300-50 or alike). This C4 material is packed into 50 mm
injection
columns and then the metabolites are separated using a SEPBOXT"", a two
dimension
automated preparative chromatography, fractionation and collection system
(SEPIAtec
GmbH, Berlin, Germany). Standard reverse phase separation methods are used at
every level.
Level one separation is achieved using C4 (Merck LiChrospher 100 RP4e, 100A,
l5,um
or Daisogel SP-120-15-CA-P) as stationary phase (250 X 50 mm column) and a
methanol:formate buffer (0.02 M ammonium formate pH 4) gradient from 0 to 100%
methanol (17 min) followed by 100% methanol (10 min), at a flow rate of 109
mUmin.
Eluting compounds are adsorbed onto 18 C4 trap columns (Macherey & Nagel
Polygoprep RP4 300-50) strictly based on time fractionation.
In the second level of separation, each trap column is eluted individually and
compounds
are separated based on polarity using various stationary phases (250 X 25 mm
columns)
and mobile phase gradients (30 mUmin) consisting of acetonitrile (CH3CN) and
0.02 M
ammonium formate (pH 4). Table 2 summarizes the type of stationary phase and
gradients used for separating the compounds from each of the 18 trap columns.
At this
level, automated fractionation is based on UV or ELSD (evaporative light
scattering
detector) signals or time. All fractions are then freed from buffer using
solid phase
extraction (Merck LiChrolut EN, 40,um) prior to collection.
3s
19

CA 02522632 2005-10-06
Table 1. Compounds of the present invention.
CCR5 MIP-1 a Binding HIV
Compound Formula
No. AVA-No Structure Weight % Inhibition Ki (~.M) IC50 (~,M)
@ 10 ~,g/m I
0
( Id) 1168 ~ ~ 360 96 0.97
-o -
0
0
2 1170 ~ \ °" 390 28
(new)
0
0
of
3 1178 \ I \ 374 41
(new)
0
_o
i ..>,°,,,
°
(o d) 1175 ~ ~ 360 25
0
OH
OH
new 1176 / 302 38
( )

CA 02522632 2005-10-06
I
I
6 1023 \ 286 77 10.8
(old)
o d 1177 \ ~ ~ 346 44
( )
., _o
.:
HO
HO /
8 1180 \ 362 20
(new) ~oH
~o
OH
9 COMM
(old) ERCIAL ~ 286 98
w
1185 ~ ~ 270 83
(new)
0
11
(new) 1186 ~ / ~ 300 80
°
21

CA 02522632 2005-10-06
\ a~
12 ~° \ ° /
1187 I I 286 81
(new) /
Ho
O
y o o a,
13 1188 "° ~ ~ I , ~' 432 44
(new) ~ ~ I - a"
0
14 N w w
(new) 1189 ~ I ~ o ~ 313 28
OH CH3
HO
\ \CH3
O OH
15 COMM / 332 76
(old) ERCIA
H3C ~CH3
OH CH3
HO
O \ \CHs
16 COMM / 330 62
(old) ERCtA 'o
H3C ~CH3
The chemical names of compounds shown in Table 1 are as follows:
Compound No. Name
22

CA 02522632 2005-10-06
1 7~i,1 O~i-epoxy-7-hydroxy-12-methoxy-8,12-icetexadiene-
11,14-dione
2 19,20-epoxy-8-methyl-2-(1-methylethyl)-1,4-dihydroxy-3,19-
dimethoxy-4b,5,6,7,8,8a,9,10-octahydrophenanthren-10-one
3 19,20-epoxy-8-methyl-2-(1-methylethyl)-1,4-dihydroxy-3-
methoxy-4b,5,6,7,8,8a,9,10-octahydrophenanthren-10,19-
dione
4 10,14-dihydroxy-11,12-methylene ether-8,11,13-icetexatrien-
7-one
5 4b,8,8-trimethyl-1-(1-methylethyl)-4b,5,6,7,8,8a,9,10-
t 5 octahydrophenanthren-2,5-diol
6 4b,8,8-trimethyl-2-(1-methylethyl)-4b,5,6,7,8,8a,9,10-
octahydrophenanthren-3-of
7 4b,8,8-trimethyl-2-(1-methylethyl)-1,4-dihydroxy-3-methoxy-
4b,5,6,7,8,8a,9,10-octahydrophenanthren-10-one
8 8,8-dimethyl-2-(1-methylethyl)-4b-hydroxymethyl-1,4-
hydroxy-3-methoxy-4b,5,6,7,8,8a,9,10-
octahydrophenanthren-10-one
9 4b,8,8-trimethyl-1-(1-methylethyl)-4b,5,6,7,8,8a,9,10-
octahydrophenanthren-2-of
10 6,7-dihydroxy-2-(4-hydroxyphenyl)-4H-chromen-4-one
11 6,7,8-trihydroxy-2-(4-methoxyphenyl)-4H-chromen-4-one
12 6,7,8-trihydroxy-2-(4-hydroxyphenyl)-4H-chromen-4-one
23

CA 02522632 2005-10-06
13 6,7,8-trihydroxy-2-(4-glucopyranosylphenyl)-4H-chromen-4-
one
14 (2E)-N[2-(4-hydroxy-2-methoxyphenyl)ethyl]-3-(4-
hydroxyphenyl)acrylyamide
5,6-dihydroxy-1,1-dimethyl-7-propan-2-yl-2,3,4,9,10,1 Oa-
hexahydrophenanthrene-4a-carboxylic acid
10 16 5,6-dihydroxy-1,1-dimethyl-7-(1-methylethyl)-1,3,4,9,10,1 Oa-
hexahydro-4a-(epoxymethano)phenanthren-12-one
15 Table 2. General separation methodology. In all cases solvents were CH3CN
and 0.02 M
ammonium formate buffer.
Trap Stationary Mobile phase compositionMobile phase composition


# Phase METHOD A METHOD B


1 NA NA


k
C8
M


;
erc


2 LiChrospher NA NA
60


3 RP-Select NA NA
B,


12,um


4 NA NA


5 NA NA


6 NA NA


7 YMC ODS-A NA NA


g Cl8e, 120A, NA NA


1 1
um


, NA NA


10 0 min 24% CH3CN 0 min 24% CH3CN


50 min 41 % CH3CN 80 min 41 % CH3CN


60 min 100% CH3CN 90 min 100% CH3CN


11 0 min 28% CH3CN 0 min 28% CH3CN


50 min 51% CH3CN 80 min 51% CH3CN


60 min 100% CH3CN 90 min 100% CH3CN


12 NA NA


13 Merck NA NA


24

CA 02522632 2005-10-06
14 LiChrospher 0 min 35% CH3CN 0 min 35% CH3CN


RP18, l2,um 50 min 70% CH3CN 80 min 70% CH3CN


60 min 100% CH3CN. 90 min 100% CH3CN


15 0 min 50% CH3CN 0 min 50% CH3CN


50 min 98% CH3CN 80 min 98% CH3CN


60 min 100% CH3CN 90 min 100% CH3CN


16 NA NA


17 NA NA


18 NA NA


NA = not applicable
In the following, the references to Compound nos. [1] to [16] are to be
understood as
respectively being a reference to a so designated compound numbered as set
forth in
Table 1 above.
Compound no. [1]:
Purification
Initial fractionation (SEPBOXT"") of Hernandia ovigera extract as described
led to two
sets of fractions with CCR5 binding inhibitory activity. The first active
fraction eluted with
100% methanol at 20 minutes (Trap column #14) in level one separation and was
eluted
between 61 and 67 minutes in level two separation using method B (Table 2).
The
second active fraction also eluted with 100% methanol at 21 minutes (Trap
column #15)
in level one separation and subsequently eluted between 30 and 36 minutes in
level two
separation using method B (Table 2 above).
Further separations by reverse phase C18 semi-preparative chromatography (150
X
21.1 mm column packed with Phenomenex Luna C18(2), 100A, l0,um) using an
acetonitrile:water gradient at a flow rate of 10 mUmin (Gradient A) resulted
in CCR5
binding inhibitory fractions that eluted at 16 minutes. Active fractions with
similar LC-MS
profiles were pooled and subjected to another reverse phase C18 semi-
preparative
HPLC purification (150 X 21,1 mm column packed with Phenomenex Luna C18(2),

CA 02522632 2005-10-06
100A, 10 ,um) at 10 mUmin utilizing an acetonitrile:aqueous trifluoroacetic
acid buffer
(0.01% TFA v/v): (Gradient B) Compound no. [1] eluted at 31 minutes.
GRADIENT GRADIENT
A B


Time (min)% CH3CN % H20 Time (min) % CH3CN % Aq TFA


0 10 90 0 10 90


3 74 26 5 60 40


30 76 24 45 62 38


31 100 0 46 100 0


35 100 0 53 100 0


36 10 90 54 10 90


41 10 90 60 10 90


Structure determination:
NMR data for compound no. [1]
'H NMR (CDCI3, 500 MHz) 8 0.86 (3H, s, CH3-19), 0.95 (3H, s, CH3-18), 1.15 (1
H, m, H-
3a), 1.21 (6H, d, J=7.0 Hz, CH3-16 and 17), 1.59 (1 H, m, H-3~), 1.60 (2H, m,
H-1 ), 1.71
(1 H, t, J-- 8.2 Hz, H-5), 1.73 (1 H, m, H-2a), 2.01 (1 H, m, H-2~), 2.04 (1
H, dd, J=12.8, 8.0
Hz, H-6(3), 2.25 (1 H, d, J= 19.5 HZ, H-20a), 2.30 (1 H, dd, J--13.0, 8.5 Hz,
H-6a), 2.55
(1 H, d, J--19.5 Hz, H-20~i), 3.23 (1 H, sep, J--7.0, H-15), 3.98 (3H, s,
OCH3).
'3C NMR (CDCI3, 500 MHz) S 15.87, 20.64, 20.68, 24.56, 27.32, 29.98, 30.54,
31.37,
32.25, 39.23, 45.96, 51.66, 61.43, 79.60, 101.13, 137.31, 139.16, 142.32,
156.34,
183.81, 189.36.
The NMR data and the molecular weight suggested the structure of compound no.
[1]
this was identical to that described (Uchiyama, N. et al., J. Nat. Prod. 2003,
66, 128-
131 ). This compound no. 1 was previously known as showing trympanocidal
activity
Compound no. [2]:
Purification
An extract of Hernandia ovigera prepared as for Compound no. [1] was dried
under in
vacuo. The resulting material was suspended in water and extracted twice with
ethyl
acetate. The combined organic fraction was subjected to manual low pressure
C18
26

CA 02522632 2005-10-06
reverse phase flash chromatography (SiliCycle RP-C18, 60A, 40-63,um) using a
gradient-like sequence of discrete mobile phase volumes composed of
acetonitrile and
water (FLASH 1 ). The fraction eluted at 100% CH3CN was subjected to reverse
phase
C18 semi-preparative HPLC chromatography purification (150 X 2l,imm column
packed
with Phenomenex Luna C18(2), 100A, l0,um) utilizing an acetonitrile:aqueous
trifluoroacetic acid buffer (0.01 % TFA v/v) at 15 mUmin (Gradient C).
Compound no.
[2] eluted at 55 minutes.
FLASH 1 GRADIENT
C


CH3CN Vol (mL) Time % CH3CN % TFA


5 525 0 10 90


20 675 5 50 50


30 600 87 18 82


35 250 88 0 100


45 200 94 0 100


55 350 95 90 10


100 1000 100 90 10



Structure determination:
NMR data for compound no. [2]
'H NMR (CDCI3, 500 MHz) 8 0.97 (3H, s, CH3-18), 1.31-1.41 (2H, m, H-ia, H3a),
1.37 (3H, d, J--4.5 Hz, CH3-16), 1.39 (3H, d, J=4.5 Hz, CH3-17), 1.62 (3H, m,
H-
3~3 and H-2), 1.87 (1 H, dd, J= 15.2, 2.5 Hz, H-5), 2.67 (1 H, dd, J=16.5, 3.5
Hz, H-
6a), 3.29 (3H, s, OCH3), 3.30 (2H, m, H-6~ and H-15), 3.42 (1 H, dd, J=9.5,
3.5
Hz, H-1 a), 3.78 (3H, s, OCH3), 3.93 (1 H, dd, J= 11.5 Hz, H-20a), 4.28 (1 H,
s, H-
19), 4.36 (1 H, d, J-- 11.5 Hz, H-20~), 5.69 (1 H, s, OH), 13.04 (1 H, s, OH).
'3C NMR (CDC13, 500 MHz) S 20.51, 20.54, 21.93, 23.50, 26.29, 35.00, 36.59,
37.60, 39.03, 39.71, 44.54, 55.05, 59.84, 62.33, 105.60, 113.37, 127.20,
128.50,
140.07, 152.22, 157.93, 205.65.
27

CA 02522632 2005-10-06
This NMR was similar to that previously decribed in Uchiyama, N. et al., J.
Nat. Prod.,
2003, 66, 128-131. The presence of the ester linkage is consitent with the
above data
as confimed by comparison with chemical shifts of compounds isolated from
Salvia
(Esquivel et al. Phytochem. 1997 46: 531-534). The structure of compound no.
[2] was
confirmed by 2D NMR (Figure 1 to 3).
Compound no. [3]:
Purification
An extract of Hernandia ovigera cell culture was prepared and subjected to low
pressure
reverse phase chromatography as described for compound 2 (FLASH 1 ). Material
eluted with 100 CH3CN was purified (8 runs) over polar end-capped C18 reverse
phase
preparative HPLC (250 X 25 mm column packed with Phenomenex Synergi
Fusion)using acetonitrile:water at mUmin. (GRADIENT D): Fractions eluted at
about
64:36 acetonitrile:water were pooled and further purified through reverse
phase C18
semi-preparative HPLC (150 X 21,1mm column packed with Phenomenex Luna C18(2),
100A, l0,um) using a methanol:water gradient at 10 mUmin: (GRADIENT E):
Compound no. [3] eluted at 53 minutes.
GRADIENT GRADIENT E
D


Time % CH3CN % H20 Time % MeOH % H20


0 10 90 0 10 90


15 60 40 3 10 90


180 90 10 15 65 35


195 100 0 65 70 30


70 100 0


75 100 0


78 10 90


80 10 90


Structure determination
NMR data for compound no. [3]
28

CA 02522632 2005-10-06
'H NMR (CDC13, 500 MHz) 8 1.30 (3H, s, CH3-18), 1.38 (3H, d, J=3.0 Hz, CH3-
17), 1.39
(3H, d, J--3.0 Hz, CH3-16), 1.47 (1 H, m, H-1 a), 1.64 (2H, m, H-3), 1.93 (2H,
m, H-2),
2.20 (1 H, dd, J--13.2, 3.0 Hz, H-5), 2.52 (1 H, t, H-6a), 2.84 (1 H, dd, J=
16.9, 3.2 Hz H-
6~), 3.31 (1 H, sep, J=3.0 Hz, H-15), 3.60 (1 H, d, J= 13.5 Hz, H-1 ~), 3.81
(3H, s, OCH3),
4.57 (1 H, dd, J= 12.0 Hz, H-20a), 5.01 (1 H, d, J= 12.0 Hz, H-20(3), 5.79 (1
H, s, OH),
12.95 (1 H, s, OH).
'3C NMR (CDCI3, 500 MHz) 8 20.43, 21.27, 23.27, 26.40, 34.93, 36.91, 40.42,
42.97,
46.54, 62.54, 72.65, 112.29, 126.31, 128.44, 140.28, 152.84, 158.75, 175.43,
202.22.
The chemical shift were similar to compound no. [2], however, the differing
molecular
weights and chemicals shift suggested the presence of a lactone. This is very
similar to
compounds isolated from salvia (Esquivel et al. Phytochem. 1997 46: 531-534).
The
structure of compound no. [3], was confirmed by 2D NMR (Figure 4 to 6).
Compound no. [4]:
Purification
Initial purification of Compound 3 also afforded fractions eluting at about
76:24
acetonitrile: water. These were pooled and further purified through reverse
phase C18
semi-preparative HPLC (150 X 21,1mm column packed with Phenomenex Luna C18(2),
100A, l0,um) using a methanol:water gradient (GRADIENT F) at 10 mUmin.
Compound
no. [4] eluted at 51 minutes.
GRADIENT
F


Time % MeOH % H20


0 10 90


3 10 90


15 75 25


65 85 15


70 100 0


75 100 0


29

CA 02522632 2005-10-06
78 10 90


80 10 90


Structure determination
NMR data for compound no. [4]
'H NMR (CDCI3, 500 MHz) 8 0.86 (3H, s, CH3-19), 0.99 (3H, s, CH3-18), 1.13 (1
H, m, H-
3a), 1.27 (3H, d, J=7.5 Hz, CH3-17), 1.30 (3H, d, J=6.5 Hz, CH3-16), 1.43 (2H,
m, H-3~
and H-5), 1.53-1.57 (2H, m, H-1 a and H-2[i), 1.73 (1 H, d, ,~ 13.5 Hz, H-1
~), 1.87 (1 H,
m, H-2(3), 2.62 (1 H, d, H-6a), 2.74 (1 H, d, J= 14.0 HZ, H-20a), 3.00 (1 H,
dd, J=18.0, 9.0
Hz, H-6(3), 3.02 (1 H, d, J--14.0 Hz, H-20a), 3.40 (1 H, sep, J--7.0, H-15),
5.92 (1 H, s,
OCH20), 5.94 (1 H, s, OCH20), 13.42 (1 H, s, OH).
'3C NMR (CDC13, 500 MHz) S 18.91, 20.83, 20.96, 21.65, 24.60, 32.27, 34.24,
38.77,
40.28, 40.81, 41.60, 49.80, 49.87, 73.89, 101.03, 112.73, 114.99, 117.34,
139.49,
150.87, 160.85, 208.84.
These NMR spectra suggested an icetaxane. This compound no. [4], was identifed
as
cyclocoulterone previosly isolated by Uchiyama, N. et al., J. Nat. Prod.,
2003, 66, 128-
131.
Compound no. [5]:
Purification
Initial purification of Compound 3 also afforded fractions eluting at about
70:30
acetonitrile: water. These were pooled and further purified through reverse
phase C18
semi-preparative HPLC (150 X 21,1mm column packed with Phenomenex Luna C18(2),
100A, l0,um) using a methanol:water gradient (GRADIENT G) at 10 mUmin:
Compound
no. [5] eluted at 33 minutes.

CA 02522632 2005-10-06
GRADIENT
G


Time % MeOH % H20


0 70 30


3 70 30 5


75 25


50 85 15


70 100 0


80 100 0


m
Structure determination
NMR data for compound no. [5]
'H NMR (CDCI3, 500 MHz) S 0.92 (6H, s, CH3-18 and CH3-19), 1.23 (3H, s, CH3-
20),
r S 1.34 (3H, d, J-- 5.0 Hz, CH3-17), 1.35 (3H, d, J= 5.0 Hz, CH3-16), 2.80-
2.93 (2H, m, H-
7a and H-7[3), 3.30 (1 H, sep, J=7.0 Hz, H-15), 4.64 (1 H, s, OH), 6.51 (1 H,
d, J= 9.0 Hz,
H-11 ), 7.97 (1 H, d, J= 9.0 Hz, H-14).
'3C NMR (CDCI3, 500 MHz) 8 18.30, 19.45, 20.59, 21.45, 27.48, 28.31, 30.70,
32.90,
33.42, 39.78, 44.02, 49.00, 78.29, 114.35, 115.51, 125.96, 131.01, 143.10,
152.64.
Spectral data suggested as abietane diterpene. Which was identified as a new
compound of structure of compound no. [5]; for comparison see old compounds
described in Ying, B-P. et al., Phytochemistry, 1991, 30: 1951 and to old
compound [9].
Compound no. [6]:
Purification
Initial purification of Compound no. . [3] also afforded fractions eluting at
about 84:16
acetonitrile: water These were pooled and further purified through reverse
phase C18
semi-preparative HPLC (150 X 21,1mm column packed with Phenomenex Luna C18(2),
100A, l0,um) using a methanol:water gradient (GRADIENT H) at 10 mUmin.
Compound no. [6] eluted at 29 minutes.
31

CA 02522632 2005-10-06
GRADIENT
H


Time % MeOH % H20


0 85 15


3 85 15


65 95 5 -'


75 100 0


80 100 0


Structure determination
NMR data for compound no. [6]
'H NMR (CDCI3, 500 MHz) 8 0.93 (3H, s, CH3-19), 0.95 (3H, s, CH3-18), 1.18
(3H, s,
CH3-20), 1.24 (3H, d, J= 7.0 Hz, CH3-17), 1.26 (3H, d, J-- 7.0 Hz, CH3-16),
2.78- 2.90
(2H, m, H-7a and H-7~i), 3.13 (1 H, sep, J--7.0 Hz, H-15), 6.64 (1 H, s, H-11
), 6.85 (1 H, s,
H-14).
'3C NMR (CDC13, 500 MHz) 8 19.48, 19.57, 21.88, 22.83, 23.02, 25.07, 27.02,
30.02,
33.58, 33.69, 37.74, 39.09, 41.90, 50.58, 111.22, 126.85, 127.52, 131.65,
148.90,
150.90.
The spectral data was consistent with ferruginol. This old compound no. [6]
has
previously been shown to possess potent antimicrobial activity.
Compound no. [7]:
Purification
From the purification of Compound no. [6], Compound no. [7] eluted at 27
minutes.
Structure determination
NMR data for compound no. [7]
'H NMR (CDCI3, 500 MHz) 8 0.94 (3H, s, CH3-19), 0.96 (3H, s, CH3-18), 1.36
(3H, s,
CH3-20), 1.37 (3H, d, J= 6.5 Hz, CH3-17), 1.39 (3H, d, J= 6.5 Hz, CH3-16),
2.64 (2H, m,
H-6a and H-6~i), 3.32 (2H, m, H-15 and H-1 Vii), 3.78 (3H, s, OCH3), 5.70 (1
H, s, OH),
13.37 (1 H, s, OH).
32

CA 02522632 2005-10-06
'3C NMR (CDC13, 500 MHz) 8 18.12, 19.28, 20.56, 20.61, 21.80, 26.28, 33.37,
33.64,
36.11, 36.66, 40.56, 41.37, 49.87, 62.29, 112.73, 126.34, 136.11, 138.84,
152.42,
158.49, 206.42.
This NMR was identical to that described in Frontana et al., Phytochemistry,
1994, 36:
739.
Compound no. [8]
Purification
Initial purification of Compound 3 also afforded fractions eluting at about
72:28
acetonitrile: water. These fractions were pooled and further purified through
polar end-
capped C18 reverse phase preparative HPLC (250 X 25 mm column packed with
Phenomenex Synergi Fusion using a methanol:water gradient (GRADIENT I):
Compound no. [8] eluted at 53 minutes.
GRADIENT
I


Time % MeOH % H20


0 10 90


80 20


110 100 0


140 100 0


Structure determination
NMR data for compound no. [8]
'H NMR (CDCI3, 500 MHz) S 0.94 (3H, s, CH3-19), 0.98 (3H, s, CH3-18), 1.35
(3H, d, J-
6.5 Hz, CH3-17), 1.37 (3H, d, J= 6.5 Hz, CH3-16), 2.61 (1 H, dd, J=14.0, 3.0
Hz, H-6a),
2.79 (1 H, t, J=15.0 Hz, H-6~), 3.38 (2H, m, H-15 and H-1 (i), 3.82 (3H, s,
OCH3), 4.04
(1 H, d, J=11.5 Hz, H-20a), 4.34 (1 H, d, J=11.5 Hz, H-20~i), 13.51 (1 H, s,
OH).
33

CA 02522632 2005-10-06
1~C NMR (CDC13, 500 MHz) S 19.14, 20.62, 20.73, 22.35, 25.84, 32.46, 33.46,
33.68,
35.75, 41.10, 45.83, 50.05, 61.96, 65.26, 113.05, 127.87, 132.35, 140.47,
154.10,
159.19, 205.77.
The spectral data points to a structure as compound no. [8] ; comfirmed by 2D
NMR
spectroscopy (Figure 7 to 9).
Compound no. [9] (Totarol), no. [15] (carnosic acid), and no. [16] (carnasol)
Compound nos. [9], [15], and [16] are commercially available plant natural
products.
Totarol was first isolated from the bark of Podocarpus nagi (Podocarpaceae) as
an
antimicrobial agent (Kubo I, Muroi H, and Himejima M (1992) J. Nat. Prod. 55,
1436-40).
Carsosic acid is an anti-oxidant. (Munne-Bosch a tal. 1999 Plant Physiol 121:
1047-
1052). Carnosol also shows anti-oxidant activity (Bozan et al., Chemistry of
Natural
Compounds, 2002, 38, 198-200) and has anti-proliferative properties (Costa-
Lotufo,et
al., Pharmazie 2004 59 :78-79).
Up until the present invention no CCR5 antagonist activity has been described
for
compound [9], [15] or [16].
Compound nos. [10 tol4]
Active fractions identified from Oreocereus bruennowii are treated in the same
way as
Hernandia ovigera above so as to isolate compound nos. [10 to 14].
Compound no. [10]:
Upon general preparative fractionation (SEPBOXT"") of Oreocereus bruennowii
methanol
plant cell culture extract, active fractions containing a mixture of compound
nos. [10],
[11], and [14] eluted at about 92% methanol after 17 minutes (Trap column #11)
in level
one separation and eluted subsequently between 35 and 45 minutes in level two
separation using the appropriate 60-minute long method (Table "Z"). These
fractions
were pooled and subjected to C12 reverse phase preparative HPLC purification
(250 X
25 mm column packed with Phenomenex Synergi Max RP) using a methanol:aqueous
trifluoroacetic custom method (GRADIENT J) at 10 mUmin to afford compound nos.
34

CA 02522632 2005-10-06
[10], [11] and [14]. Compound no. [10] eluted as a broad peak between 38 and
42
minutes, Compound no. [11] eluted between 44 and 50 minutes, and Compound no.
[14] eluted between 26 and 29 minutes
GRADIENT
J


Time % MeOH % Aq
TFA


0 10 90


20 50 50


105 50 50


120 100 0


Structure determination
NMR data for compound no. [10]
' H NMR (CDCI3, 500 MHz) 8 6.60 (1 H, s, H-3), 6.71 (1 H, s, H-5), 7.55 (2H,
d, J= 7.5 Hz,
H2' and H6'), 7.56 (1 H, s, H-8), 7.97 (2H, d, ,~ 7.5 Hz, H3' and H5').
isC NMR (CDCI3, 500 MHz) 8 93.86, 104.18, 104.67, 126.19, 129.03, 129.55,
131.55,
131.74, 150.97, 153.72, 164.38, 183.02.
The spectral data pointed to the simple flavanoid of structure as for compound
no. [10].
This was confirmed by 2D NMR spectroscopy.
Compound [11]
Purification
Compound no. [11] was isolated as describe for Compound no. [10].
Structure determination
NMR data for compound no. [11]
'H NMR (CDCI3, 500 MHz) 8 3.89 (3H, s, OCH3), 6.58 (1 H, s, H-3), 6.61 (1 H,
s, H-5),
7.07 (2H, d, J-- 8.5 Hz, H2' and H6'), 7.92 (2H, d, ,~ 8.5 Hz, H3' and H5')
The spectral data pointed to a simple flavanoid with a methoxy substitution
and a

CA 02522632 2005-10-06
hydroxyl of structure as for compound no. [11].
Compound no. [12]:
Upon general preparative fractionation (SEPBOXT"") of Oreocereus bruennowii
methanol
plant cell culture extract, the active fraction containing compound nos. [12]
and [13]
eluted at 86% methanol after 16 minutes (Trap column #10) in level one
separation and
eluted subsequently between 16 and 18 minutes in level two separation using
method A
(Table "Z"). This fraction was further purified through reverse phase C18 semi-

preparative HPLC (150 X 21,1mm column packed with Phenomenex Luna C18(2),
100A, l0,um) using a methanol:aqueous trifluoroacetic gradient (GRADIENT K) at
10
ml/min. Compound no. [12] eluted at 26 minutes and Compound no. [13] eluted at
23
minutes.
GRADIENT
K


Time % MeOH % Aq. TFA


0 10 90


3 10 90


10 45 55


50 55 45


51 100 0


60 100 0


Structure determination
NMR data for compound no. [12]
'H NMR (CDCI3, 500 MHz) b 6.58 (1 H, s, H-3), 6.59 (1 H, s, H-5), 6.92 (2H, d,
J= 8.5 Hz,
H2' and H6'), 7.84 (2H, d, ,~ 8.5 Hz, H3' and H5')
The spectral data for compound no. [12] was almost identical to that of
compound no.
[11] except the lack of the methoxy group.
Compound no. [13]
Compound no. [13] was isolated as described for Compound no. [12].
36

CA 02522632 2005-10-06
Structure determination
NMR data for compound [13]
'H NMR (CDCI3, 500 MHz) b 3.44-3.99 (6H, m, H-2', 3', 4', 5' and 6'), 5.08 (1
H, d, J-- 7.5
Hz, H-1'), 6.79 (1 H, s, H-3), 7.06 (1 H, s, H-5), 7.56 (2H, d, J-- 7.5 Hz,
H2' and H6'), 8.03
(2H, d, J-- 7.5 Hz, H3' and H5')
The'H-NMR signals at 3.44-3.99 suggest the presence of a glycoside. The
remaining
chemical shifts indicated that it was a 4' glycosyl of compound no. [10].
Compound 14:
Purification
Compound no. [14] was isolated as described for Compound no. [10].
Structure determination
NMR data for compound no. [14]
'H NMR (CDC13, 500 MHz) 8 2.74 (2H, t, J= 6.7 Hz, H-T), 3.48 (2H, t, J= 6.7
Hz, H-8'),
3.81 (3H, s, OCH3), 6.58 (1 H, d, u'= 16.5 Hz, H-7), 6.67 (1 H, d, J-- 7.5 H-
5'), 6.72 (1 H, s,
H-3'), 6.84 (1 H, d, J=8.0 Hz, H-6'), 7.37 (2H, d, J= 7.0 Hz, H2' and H6'),
7.52 1 H, d, J=
15.5 Hz, H-8), 7.54 (2H, d, J-- 7.0 Hz, H3' and H5').
'3C NMR (CDC13, 500 MHz) b 34.78, 41.28, 48.69, 55.28, 111.74, 115.63, 119.78,
120.69, 127.64, 128.77, 129.63, 132.23, 135.12, 140.47, 146.34, 146.49,
167.43.
The spectral data for compound no. [14] was consistent with an amide with two
hydroxyls and a methoxy. 13C and 2D NMR confirmed this structure.
General Structure
37

CA 02522632 2005-10-06
The present invention further provides compounds of the generic structures
which are
described below.
From the above examples, it is expected that Compounds of the generic
structures
which follow, should exhibit CCR5 activity, namely Icetaxanes [17, 20],
abietataxanes
[18, 19, 19a, 19b], flavanoids [21] and cinnamoyl amides [22].
The compounds of the following generic structures may comprise groups such as
hydroxyl, alkoxy, alkyl, halo, haloalkyl, alkylcarbonyloxy, etc., as well as
(aliphatic,
cyclic, aromatic or basic) amino acid residues. In accordance with the present
invention,
an alkyl group or moiety may be straight or branched and may itself be
unsubstitiuted or
substituted by a substituent group. Thus, for example a compound of a generic
formula
which follows may comprise an aminoalkylcarbonyloxy group. The alkyl moiety of
an
aminoalkylcarbonyloxy group may be straight or branched (e.g. be an isopropyl
group)
and may itself be unsubstituted or substituted; if substituted an alkyl moiety
may, for
example, be substituted by one or more amino groups. Thus an
aminoalkylcarbonyloxy
group may for example be amino-(C1_3alkyl)-COO-, and in particular an amino
acid
residue of formula N(R4)(R5)-(C1-C3-alkyl)carbonyl wherein R4 and R5 may for
example be independently selected from the group consisting of -H, C,-C3-
alkyl, C1-C$-
alkylcarbonyl, etc.; more particularly an alpha amino acid residue of formula
R-CH(NH2)COO-
wherein R may be a substituent group of any (known) essential or alpha amino
acid and
may have the example values as set forth below:
38

CA 02522632 2005-10-06
COMMON a-AMINO ACIDS, R-CH(NH2)C02H
ISOELECTRIC


NAME (ABBREVIATION) R POINT (pI)


glycine (gly) H- 5.97


alanine (ala ) 6.02
CH3-


valine (val) (CH3)2CH- 5.97


leucine (leu) (CH3)QCHCHZ- 5.98


isoleucine (ile, ileu)CH3CH2CH(CH3)- 6.02


H


prolinea (pro) N 6.10
COzH


H
%


N


hydroxyproline~ (hypro)Cp2H 5.78


HO


phenylalanine (phe) CsHsCH2- 5.88


CH2-



tryptophan (try)
5.88


N
H


methionine (met) CH3SCHzCHz- 5.75


aspartic acid (asp) H02CCH2- 2.87


asparagine (asp(NH2) HzlVC(O)CHZ 5.41
or asn)


glutamic acid (glu) HOZCCHZCH2- 3:22


glutamine (glu(NHZ) HzNC(O)CH2CHz 5.65
or gln)


lysine (lys) HZNCHzCH2CH2CH2- 9.74


HN


\


arginine (arg) C-NH-CH2CH2CH2- 10.76


H2N


N


histidine (his) C~ 7.58
~


N
CH2-
H


serine (ser) HOCH2 5.68


threonine (thr) CH3CH(OH)- 5.08


' /
\


tyrosine (tyr) HO 5.65
CH2-


cysteine (cySH) HSCHZ- 5.02


cystine (cyS-Scy) -CHZS-SCH2- 5.06


The present invention further includes esters compounds which may operate as
prodrugs, hydrolyzing under normal physiological conditions to provide
therapeutically
active compounds.
39

CA 02522632 2005-10-06
Thus the present invention relates to the following compounds, namely:
a compound of formula 17
E
17
where R1 and Rya together represent O or alternatively Rya is H and R1 is
selected from
the group consisting of -OH, alkoxy such as for example alkoxy having from 1
to 3
carbon atoms ((i.e. C~_3alkyl-O-, e.g. CH3-O-), halogen (e.g. F, CI, or Br,)
haloalkyl
such as for example haloalkyl having from 1 to 3 carbon atoms i.e. halo-
C1_3alkyl, (e.g.
fluro-C,_3alkyl, e.g. -CF3), alkylcarbonyloxy such as for example
alkylcarbonyloxy having
from 1 to 3 carbon atoms C1_3alkyl-COO-, (e.g. Acetyl0-);
where R2 is selected from the group consisting of -OH, alkoxy such as for
example
alkoxy having from 1 to 3 carbon atoms ((i.e. C,_3alkyl-O-, e.g. CH3-O-),
halogen (e.g.
F, CI, or Br,) haloalkyl such as for example haloalkyl having from 1 to 3
carbon atoms
i.e. halo-C,_3alkyl, (e.g. fluro-C1_3alkyl, e.g. -CF3), alkylcarbonyloxy such
as for example
C,_3alkyl-COO-, (e.g. Acetyl0-);
where R3 and R3a together represent O or alternatively R3a is H and R3 is
selected from
the group consisting of -OH, alkoxy such as for example alkoxy having from 1
to 3
carbon atoms ((i.e. Ci_3alkyl-O-, e.g. CH3-O-), halogen (e.g. F, CI, or Br,)
haloalkyl
R. Ria r,

CA 02522632 2005-10-06
such as for example halo-C~_3alkyl, (e.g. fluro-C,_3alkyl, e.g. -CF3),
alkylcarbonyloxy
such as for example C,_3alkyl-COO-, (e.g. -OAcetyl);
where R~, is selected from the group consisting of -OH, alkoxy such as for
example
example alkoxy having from 1 to 3 carbon atoms ((i.e. C,_3alk-O-,( e.g. OCH3),
halogen
(e.g. F, CI, or Br,) haloalkyl such as for example halo-C,_3alkyl, (e.g. fluro-
Ci_3alkyl, e.g.
-CF3), alkylcarbonyloxy such as for example C,_3alkyl-COO-, (e.g. Acetyl0-);
a compound of formula 18
Rs CH3
R,
0
I -R$
~~CHg R8a
Rsa 1
Rs
18
H3
where R5 is selected from the group consisting of -OH, alkoxy such as for
example
alkoxy having from 1 to 3 carbon atoms ((i.e. C,_3alkyl-O-, e.g. CH3-O-),
halogen (e.g.
F, CI, or Br,) haloalkyl such as for example haloalkyl having from 1 to 3
carbon atoms
i.e. halo-Ci_3alkyl, (e.g. fluro-C,_3alkyl, e.g. -CF3), alkylcarbonyloxy such
as for example -
C,_3alkyl-COO-, (e.g. Acetyl0-);
where R6 is selected from the group consisting of -OH, alkoxy such as for
example
alkoxy having from 1 to 3 carbon atoms ((i.e. C,_3alkyl-O-, e.g. CH3-O-),
halogen (e.g.
F, CI, or Br,) haloalkyl such as for example haloalkyl having from 1 to 3
carbon atoms
41

CA 02522632 2005-10-06
i.e. halo-C1_3alkyl, (e.g. fluro-Ci_3alkyl, e.g. -CF3), alkylcarbonyloxy such
as for example
C1_3alkyl-COO-, (e.g. Acetyl0-);
where R, is selected from the group consisting of -OH, alkoxy such as for
example
alkoxy having from 1 to 3 carbon atoms ((i.e. Ci_3alkyl-O-, e.g. CH3-O-),
halogen (e.g.
F, CI, or Br,) haloalkyl such as for example haloalkyl having from 1 to 3
carbon atoms
i.e. halo-Ci_3alkyl, (e.g. fluro-C1_3alkyl, e.g. -CF3), alkylcarbonyloxy such
as for example -
C1_3alkyl-COO-, (e.g. Acetyl0-);
where R$ and Rsa together represent O or alternatively Rsa is H and R$ is
selected from
the group consisting of -OH, alkoxy such as for example alkoxy having from 1
to 3
carbon atoms ((i.e. C1_3alkyl-O-, e.g. CH3-O-), halogen (e.g. F, CI, or Br,)
haloalkyl
such as for example halo-C1_3alkyl, (e.g. fluro-C1_3alkyl, e.g. -CF3),
alkylcarbonyloxy
such as for example Ci_3alkyl-COO-, (e.g. -OAcetyl);
where R9 and R9a together represent O or alternatively R9a is H and R9 is
selected from
the group consisting of -OH, alkoxy such as for example alkoxy having from 1
to 3
carbon atoms ((i.e. C1_3alkyl-O-, e.g. CH3-O-), halogen (e.g. F, CI, or Br,)
haloalkyl
such as for example halo-Ci_3alkyl, (e.g. fluro-C1_3alkyl, e.g. -CF3),
alkylcarbonyloxy
such as for example C1_3alkyl-COO-, (e.g. -OAcetyl).
compounds of formula 19, 19a and 19b
R,~ R1s
R14 R12 ~ R14
R10
R11
R15 ~~ \R15
I R1s
i Rl6a
H~C CHs
9
19 19a
42

CA 02522632 2005-10-06
R13
R12 ~ R14
R11
w
R15
Ris
CH3 Risa
19b
5 where Rio is selected from the group consisting of H, -OH, alkoxy such as
for example
alkoxy having from 1 to 3 carbon atoms ((i.e. C1_3alkyl-O-, e.g. CH3-O-),
halogen (e.g.
F, CI, or Br,) haloalkyl such as for example haloalkyl having from 1 to 3
carbon atoms
i.e. halo-Cl.3alkyl, (e.g. fluro-Cl.3alkyl, e.g. -CF3), alkylcarbonyloxy such
as for example
Ci_3alkyl-COO-, (e.g. Acetyl0-);
where R11 is selected from the group consisting of H, -OH, alkoxy such as for
example
alkoxy having from 1 to 3 carbon atoms ((i.e. C1_3alkyl-O-, e.g. CH3-O-),
halogen (e.g.
F, CI, or Br,) alkyl such as for example alkyl having from 1 to 3 carbon atoms
i.e.
C1_3alkyl, hydroxyalkyl such as for example hydroxyalkyl having from 1 to 3
carbon
atoms i.e. HO-C1_3alkyl, (e.g. hydroxyl-CH2-, 2-hydroxy-ethyl, 1-hydroxy-
ethyl, etc.),
haloaikyl such as for example haloalkyl having from 1 to 3 carbon atoms i.e.
halo-C1_
3alkyl, (e.g. fluro-C1_3alkyl, e.g. -CF3), alkylcarbonyloxy such as for
example Ci_3alkyl-
COO-, (e.g. Acetyl0-), carboxyl and carboxylalkyl such as for example
carboxylalkyl
having from 1 to 3 carbon atoms in the alkyl moitie i.e. HCOO-Ci_3alkyl;
where R12 is selected from the group consisting of H, -OH, alkoxy such as for
example
alkoxy having from 1 to 3 carbon atoms ((i.e. C1_3alkyl-O-, e.g. CH3-O-),
halogen (e.g.
F, CI, or Br,) alkyl such as for example alkyl having from 1 to 3 carbon atoms
i.e.
C1_3alkyl, hydroxyalkyl such as for example hydroxyalkyl having from 1 to 3
carbon
atoms i.e. HO-C1_3alkyl, (e.g. hydroxyl-CH2-, 2-hydroxy-ethyl, 1-hydroxy-
ethyl, etc.),
haloalkyl such as for example haloalkyl having from 1 to 3 carbon atoms i.e.
halo-Ci_
3alkyl, (e.g. fluro-C1_3alkyl, e.g. -CF3), alkylcarbonyloxy such as for
example Cl.3alkyl-
COO-, (e.g. Acetyl0-);
43

CA 02522632 2005-10-06
where R,3 is selected from the group consisting of H, -OH, alkoxy such as for
example
alkoxy having from 1 to 3 carbon atoms ((i.e. C1_3alkyl-O-, e.g. CH3-O-),
halogen (e.g.
F, CI, or Br,) alkyl such as for example alkyl having from 1 to 3 carbon atoms
i.e.
C,_3alkyl, hydroxyalkyl such as for example hydroxyalkyl having from 1 to 3
carbon
atoms i.e. HO-C,_3alkyl, (e.g. hydroxyl-CH2-, 2-hydroxy-ethyl, 1-hydroxy-
ethyl, etc.),
haloalkyl such as for example haloalkyl having from 1 to 3 carbon atoms i.e.
halo-C,_
3alkyl, (e.g. fluro-Ci_3alkyl, e.g. -CF3), alkylcarbonyloxy such as for
example C1_3alkyl-
COO-, (e.g. Acetyl0-);
where R~4 is selected from the group consisting of H, -OH, alkoxy such as for
example
alkoxy having from 1 to 3 carbon atoms ((i.e. C1_3alkyl-O-, e.g. CH3-O-),
halogen (e.g.
F, CI, or Br,) alkyl such as for example alkyl having from 1 to 3 carbon atoms
i.e.
C,.3alkyl, hydroxyalkyl such as for example hydroxyalkyl having from 1 to 3
carbon
atoms i.e. HO-C,_3alkyl, (e.g. hydroxyl-CH2-, 2-hydroxy-ethyl, 1-hydroxy-
ethyl, etc.),
haloalkyl such as for example haloalkyl having from 1 to 3 carbon atoms i.e.
halo-
C1_3alkyl, (e.g. fluro-C~_3alkyl, e.g. -CF3), alkylcarbonyloxy such as for
example C,_
3alkyl-COO-, (e.g. Acetyl0-);
where R,5 is selected from the group consisting of H, -OH, alkoxy such as for
example
alkoxy having from 1 to 3 carbon atoms ((i.e. C1_3alkyl-O-, e.g. CH3-O-),
halogen (e.g.
F, CI, or Br,) alkyl such as for example alkyl having from 1 to 3 carbon atoms
i.e.
C~_3alkyl, hydroxyalkyl such as for example hydroxyalkyl having from 1 to 3
carbon
atoms i.e. HO-C,_3alkyl, (e.g. hydroxyl-CH2-, 2-hydroxy-ethyl, 1-hydroxy-
ethyl, etc.),
haloalkyl such as for example haloalkyl having from 1 to 3 carbon atoms i.e.
halo-C,_
3alkyl, (e.g. fluro-Ci_3alkyl, e.g. -CF3), alkylcarbonyloxy such as for
example C1_3alkyl-
COO-, (e.g. Acetyl0-);
where R,6 and R,sa together represent O or R,6 and R,sa each represents H, or
alternatively R,sa is H and R16 is selected from the group consisting of -OH,
alkoxy such
as for example alkoxy having from 1 to 3 carbon atoms ((i.e. C~_3alkyl-O-,
e.g. CH3-O-),
halogen (e.g. F, CI, or Br,) haloalkyl such as for example halo-C,_3alkyl,
(e.g. fluro-C,_
3alkyl, e.g. -CF3), alkylcarbonyloxy such as for example C,_3alkyl-COO-, (e.g.
-OAcetyl),
or alternatively one of R,6 and R,sa represents H and the other together with
R11 defines
44

CA 02522632 2005-10-06
a bridging group of formula -COO- extending between the carbon atoms to which
they
are respective attached;
10
a compound of formula 20
RiR R...
R2o
~zza
20
where R" is selected from the group consisting of H, -OH, alkoxy such as for
example
alkoxy having from 1 to 3 carbon atoms ((i.e. C,_3alkyl-O-, e.g. CH3-O-),
halogen (e.g.
F, CI, or Br,) alkyl such as for example alkyl having from 1 to 3 carbon atoms
i.e.
C~_3alkyl, hydroxyalkyl such as for example hydroxyalkyl having from 1 to 3
carbon
atoms i.e. HO-C~_3alkyl, (e.g. hydroxyl-CH2-, 2-hydroxy-ethyl, 1-hydroxy-
ethyl, etc.),
haloalkyl such as for example haloalkyl having from 1 to 3 carbon atoms i.e.
halo-C1_
3alkyl, (e.g. fluro-C,_3alkyl, e.g. -CF3), alkylcarbonyloxy such as for
example C,_3alkyl-
COO-, (e.g. Acetyl0-);
where R,$ and Ri9 are the same or different and each is selected from the
group
consisting of H, -OH, alkoxy such as for example alkoxy having from 1 to 3
carbon
atoms ((i.e. C1_3alkyl-O-, e.g. CH3-O-), halogen (e.g. F, CI, or Br,) alkyl
such as for
example alkyl having from 1 to 3 carbon atoms i.e. C,_3alkyl, hydroxyalkyl
such as

CA 02522632 2005-10-06
for example hydroxyalkyl having from 1 to 3 carbon atoms i.e. HO-C1_3alkyl,
(e.g.
hydroxyl-CH2-, 2-hydroxy-ethyl, 1-hydroxy-ethyl, etc.), haloalkyl such as for
example
haloalkyl having from 1 to 3 carbon atoms i.e. halo-C,_3alkyl, (e.g. fluro-
C,_3alkyl, e.g. -
CF3), alkylcarbonyloxy such as for example C1_3alkyl-COO-, (e.g. Acetyl0-); or
alternatively R,$ and R~9 together define a bridging group of formula -OCH20-
so as to
form, with the carbon atom to which they are respective attached, a 5 member
heterocyclic ring;
where R2o is selected from the group consisting of H, -OH, alkoxy such as for
example
alkoxy having from 1 to 3 carbon atoms ((i.e. C1_3alkyl-O-, e.g. CH3-O-),
halogen (e.g.
F, CI, or Br,) alkyl such as for example alkyl having from 1 to 3 carbon atoms
i.e.
C,.3alkyl, hydroxyalkyl such as for example hydroxyalkyl having from 1 to 3
carbon
atoms i.e. HO-C,_3alkyl, (e.g. hydroxyl-CH2-, 2-hydroxy-ethyl, 1-hydroxy-
ethyl, etc.),
haloalkyl such as for example haloalkyl having from 1 to 3 carbon atoms i.e.
halo-Ci_
3alkyl, (e.g. fluro-C~_3alkyl, e.g. -CF3), alkylcarbonyloxy such as for
example C1_3alkyl-
COO-, (e.g. Acetyl0-);
where R2, is selected from the group consisting of H, -OH, alkoxy such as for
example
alkoxy having from 1 to 3 carbon atoms ((i.e. C~_3alkyl-O-, e.g. CH3-O-),
halogen (e.g.
F, CI, or Br,) alkyl such as for example alkyl having from 1 to 3 carbon atoms
i.e.
C,_3alkyl, hydroxyalkyl such as for example hydroxyalkyl having from 1 to 3
carbon
atoms i.e. HO-C,_3alkyl, (e.g. hydroxyl-CH2-, 2-hydroxy-ethyl, 1-hydroxy-
ethyl, etc.),
haloalkyl such as for example haloalkyl having from 1 to 3 carbon atoms i.e.
halo-C,_
3alkyl, (e.g. fluro-C1_3alkyl, e.g. -CF3), alkylcarbonyloxy such as for
example C,_3alkyl-
COO-, (e.g. Acetyl0-);
where R22 and R22a together represent O or R22 and R22a each represents H, or
alternatively R,sa is H and R,s is selected from the group consisting of -OH,
alkoxy such
as for example alkoxy having from 1 to 3 carbon atoms ((i.e. C~_3alkyl-O-,
e.g. CH3-O-),
halogen (e.g. F, CI, or Br,) haloalkyl such as for example halo-C1_3alkyl,
(e.g. fluro-C~_
3alkyl, e.g. -CF3), alkylcarbonyloxy such as for example C,_3alkyl-COO-, (e.g.
-OAcetyl).
46

CA 02522632 2005-10-06
a compound of formula 21
R26
O
21
where R23 is selected from the group consisting of -OH, alkoxy such as for
example
alkoxy having from 1 to 3 carbon atoms ((i.e. C1_3alkyl-O-, e.g. CH3-O-),
halogen (e.g.
F, CI, or Br,) alkyl such as for example alkyl having from 1 to 3 carbon atoms
i.e.
C,_3alkyl, hydroxyalkyl such as for example hydroxyalkyl having from 1 to 3
carbon
atoms i.e. HO-C1_3alkyl, (e.g. hydroxyl-CH2-, 2-hydroxy-ethyl, 1-hydroxy-
ethyl, etc.),
haloalkyl such as for example haloalkyl having from 1 to 3 carbon atoms i.e.
halo-C,_
3alkyl, (e.g. fluro-Ci_3alkyl, e.g. -CF3), alkylcarbonyloxy such as for
example C1_3alkyl-
COO-, (e.g. Acetyl0-);
where R24 is selected from the group consisting of -OH, alkoxy such as for
example
alkoxy having from 1 to 3 carbon atoms ((i.e. C,-salkyl-O-, e.g. CH3-O-),
halogen (e.g.
F, CI, or Br,) alkyl such as for example alkyl having from 1 to 3 carbon atoms
i.e.
C1_3alkyl, hydroxyalkyl such as for example hydroxyalkyl having from 1 to 3
carbon
atoms i.e. HO-C,_3alkyl, (e.g. hydroxyl-CH2-, 2-hydroxy-ethyl, 1-hydroxy-
ethyl, etc.),
haloalkyl such as for example haloalkyl having from 1 to 3 carbon atoms i.e.
halo-C~_
3alkyl, (e.g. fluro-C1_3alkyl, e.g. -CF3), alkylcarbonyloxy such as for
example C,_~alkyl-
COO-, (e.g. Acetyl0-);
where R25 is selected from the group consisting of H, -OH, alkoxy such as for
example
alkoxy having from 1 to 3 carbon atoms ((i.e. C,_3alkyl-O-, e.g. CH3-O-),
halogen (e.g.
47

CA 02522632 2005-10-06
F, CI, or Br,) alkyl such as for example alkyl having from 1 to 3 carbon atoms
i.e.
C,_3alkyl, hydroxyalkyl such as for example hydroxyalkyl having from 1 to 3
carbon
atoms i.e. HO-C,.3alkyl, (e.g. hydroxyl-CH2-, 2-hydroxy-ethyl, 1-hydroxy-
ethyl, etc.),
haloalkyl such as for example haloalkyl having from 1 to 3 carbon atoms i.e.
halo-C~_
alkyl, (e.g. fluro-C,_3alkyl, e.g. -CF3), alkylcarbonyloxy such as for example
C~_3alkyl-
COO-, (e.g. Acetyl0-);
where R26 is selected from the group consisting of -OH, alkoxy such as for
example
alkoxy having from 1 to 3 carbon atoms ((i.e. C1_3alkyl-O-, e.g. CH3-O-),
halogen (e.g.
F, CI, or Br,) alkyl such as for example alkyl having from 1 to 3 carbon atoms
i.e.
Ci_3alkyl, hydroxyalkyl such as for example hydroxyalkyl having from 1 to 3
carbon
atoms i.e. HO-Cl.3alkyl, (e.g. hydroxyl-CH2-, 2-hydroxy-ethyl, 1-hydroxy-
ethyl, etc.),
haloalkyl such as for example haloalkyl having from 1 to 3 carbon atoms i.e.
halo-C,_
3alkyl, (e.g. fluro-Ci_3alkyl, e.g. -CF3), alkylcarbonyloxy such as for
example C1_3alkyl-
COO-, (e.g. Acetyl0-); an oxygen linked carbohydrate residue, namely a
carbohydrate
residue linked via an oxygen of an hydroxyl group of the carbohydrate to the
phenyl
group (e.g. a monosaccharide residue, a disaccharide residue, a trisaccharide
residue or
other glycoside residue which yields a sugar substance on hydrolysis, e.g. a O-
linked
residue of glucose, fructose, lactose, mannose, galactose, etc. which on
hydrolysis
yields glucose, fructose, lactose, etc..)
30
a compound of formula 22
48

CA 02522632 2005-10-06
\ Rso
NH
\ ~/ ~/ ~/ ~ R3i
O
R2, ~ _ R29
R2s
22
where R2, is selected from the group consisting of -OH, alkoxy such as for
example
alkoxy having from 1 to 3 carbon atoms ((i.e. Cl.3alkyl-O-, e.g. CH3-O-),
halogen (e.g.
F, CI, or Br,) alkyl such as for example alkyl having from 1 to 3 carbon atoms
i.e.
C~_3alkyl, hydroxyalkyl such as for example hydroxyalkyl having from 1 to 3
carbon
atoms i.e. HO-C,_3alkyl, (e.g. hydroxyl-CH2-, 2-hydroxy-ethyl, 1-hydroxy-
ethyl, etc.),
haloalkyl such as for example haloalkyl having from 1 to 3 carbon atoms i.e.
halo-C~_
3alkyl, (e.g. fluro-C,_3alkyl, e.g. -CF3), alkylcarbonyloxy such as for
example C~_3alkyl-
COO-, (e.g. Acetyl0-);
where R28 is selected from the group consisting of H, -OH, alkoxy such as for
example
alkoxy having from 1 to 3 carbon atoms ((i.e. Ci_3alkyl-O-, e.g. CH3-O-),
halogen (e.g.
F, CI, or Br,) alkyl such as for example alkyl having from 1 to 3 carbon atoms
i.e.
C1_3alkyl, hydroxyalkyl such as for example hydroxyalkyl having from 1 to 3
carbon
atoms i.e. HO-C,_3alkyl, (e.g. hydroxyl-CH2-, 2-hydroxy-ethyl, 1-hydroxy-
ethyl, etc.),
haloalkyl such as for example haloalkyl having from 1 to 3 carbon atoms i.e.
halo-C,_
3alkyl, (e.g. fluro-C,_3alkyl, e.g. -CF3), alkylcarbonyloxy such as for
example C1_3alkyl-
COO-, (e.g. Acetyl0-);
where R29 is selected from the group consisting of -OH, alkoxy such as for
example
alkoxy having from 1 to 3 carbon atoms ((i.e. C1_3alkyl-O-, e.g. CH3-O-),
halogen (e.g.
F, CI, or Br,) alkyl such as for example alkyl having from 1 to 3 carbon atoms
i.e.
C~_3alkyl, hydroxyalkyl such as for example hydroxyalkyl having from 1 to 3
carbon
atoms i.e. HO-C1_3alkyl, (e.g. hydroxyl-CH2-, 2-hydroxy-ethyl, 1-hydroxy-
ethyl, etc.),
haloalkyl such as for example haloalkyl having from 1 to 3 carbon atoms i.e.
halo-C,_
49

CA 02522632 2005-10-06
3alkyl, (e.g. fluro-C,_3alkyl, e.g. -CF3), alkylcarbonyloxy such as for
example C1_3alkyl-
COO-, (e.g. Acetyl0-);
where R3o is selected from the group consisting of -OH, alkoxy such as for
example
alkoxy having from 1 to 3 carbon atoms ((i.e. C,.3alkyl-O-, e.g. CH3-O-),
halogen (e.g.
F, CI, or Br,) alkyl such as for example alkyl having from 1 to 3 carbon atoms
i.e.
C1_3alkyl, hydroxyalkyl such as for example hydroxyalkyl having from 1 to 3
carbon
atoms i.e. HO-C,_3alkyl, (e.g. hydroxyl-CH2-, 2-hydroxy-ethyl, 1-hydroxy-
ethyl, etc.),
haloalkyl such as for example haloalkyl having from 1 to 3 carbon atoms i.e.
halo-C,_
3alkyl, (e.g. fluro-C,_3alkyl, e.g. -CF3), alkylcarbonyloxy such as for
example Ci_3alkyl-
COO-, (e.g. Acetyl0-);
where R31 is selected from the group consisting of H, -OH, alkoxy such as for
example
alkoxy having from 1 to 3 carbon atoms ((i.e. C1_3alkyl-O-, e.g. CH3-O-),
halogen (e.g.
F, CI, or Br,) alkyl such as for example alkyl having from 1 to 3 carbon atoms
i.e.
C,_3alkyl, hydroxyalkyl such as for example hydroxyalkyl having from 1 to 3
carbon
atoms i.e. HO-C,_3alkyl, (e.g. hydroxyl-CH2-, 2-hydroxy-ethyl, 1-hydroxy-
ethyl, etc.),
haloalkyl such as for example haloalkyl having from 1 to 3 carbon atoms i.e.
halo-C1_
3alkyl, (e.g. fluro-C,_3alkyl, e.g. -CF3), alkylcarbonyloxy such as for
example C,_3alkyl-
COO-, (e.g. Acetyl0-);
Compounds of the above generic formulae may, for example, be obtained starting
from
the compounds set forth in table 1 above, i.e. the compounds of table 1 may be
used as
intermediate compounds for the preparation of other compound of the generic
structures
by appropriate oxidation of alcohol to carbonyl, reduction of carbonyl to
alcohol,
alkylation of , esterification etc..
Thus the synthesis of compounds derived from those in Table 1 may be achieved
in a
variety of ways. Compounds in Table 1 may be used directly for synthesis on
reactive
sites of each of the molecules. For example hydroxyls may undergo a wide
variety of
well-known reactions involving oxidation, reduction, substitution as well as
many reaction
types to afford a variety of compounds. Similar compound available from other
sources
may act as starting materials for derivatives.
50

CA 02522632 2005-10-06
Pharmaceutical compositions of the present invention may, as mentioned,
comprise any
of the compounds (including salts, prodrugs, etc.) of the present invention,
along with
any pharmaceutically acceptable carrier, adjuvant or vehicle.
The terms "pharmaceutically acceptable carrier", "pharmaceutically acceptable
adjuvant" and "physiologically acceptable vehicle" refer to a non-toxic
carrier or adjuvant
that may be administered to a patient, together with a compound of this
invention, and
which does not destroy the pharmacological activity thereof.
Pharmaceutically acceptable carriers, adjuvants and vehicles that may be used
in the
pharmaceutical compositions of this invention include, but are not limited to
ion
exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as
human serum
albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium
sorbate,
partial glyceride mixtures of saturated vegetable fatty acids, water, salts or
electrolytes,
such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen
phosphate, sodium chloride, zinc salts, colloidal silica, magnesium
trisilicate, polyvinyl
pyrrolidone, cellulose-based substances, polyethyleneglycol, sodium
carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-
block
polymers, polyethylene glycol and wool fat.
The pharmaceutical compositions of this invention may as mentioned be
administered
orally, parenterally by inhalation spray, topically, rectally, nasally,
buccally, vaginally or
via an implanted reservoir. It is therefore understood herein that oral
administration or
administration by injection are encompassed by the present invention. For
example,
compounds of the present invention, may, for example, be orally administered
in an
aqueous solution. The pharmaceutical compositions of this invention may
contain any
conventional non-toxic pharmaceutically acceptable carriers, adjuvants or
vehicles. The
term "parenteral" as used herein includes subcutaneous, intracutaneous,
intravenous,
intramuscular, intra-articular, intrasynovial, intrasternal, intrathecal,
intralesional and
intracranial injection or infusion techniques.
The pharmaceutical compositions may be in the form of a sterile injectable
preparation,
for example, as a sterile injectable aqueous or oleaginous suspension. This
suspension
may be formulated according to techniques known in the art using suitable
dispersing or
wetting agents (such as, for example, Tween 80) and suspending agents. The
sterile
51

CA 02522632 2005-10-06
injectable preparation may also be a sterile injectable solution or suspension
in a non-
toxic parenterally acceptable diluent or solvent. Among the acceptable
vehicles and
solvents that may be employed are amino acid, water, Ringer's solution and
isotonic
sodium chloride solution.
The pharmaceutical compositions of this invention may, for example, be orally
administered in any orally acceptable dosage form including, but not limited
to, capsules,
tablets, and aqueous suspension and solutions. In the case of tablets for oral
use,
carriers that are commonly used include lactose and corn starch. Lubricating
agents,
such as magnesium stearate, are also typically added. For oral administration
in a
capsule form, useful diluents include lactose and dried cornstarch. When
aqueous
suspensions are administered orally, the active ingredient is combined with
emulsifying
and suspending agents. If desired, certain sweetening and/or flavoring and/or
coloring
agents may be added.
52

Representative Drawing