WO 2006/096325 PCT/US2006/006125
OXEPANE ISOMER OF 42-O-(2-HYDROXY)ETHYL-RAPAMYCIN
BACKGROUND OF THE INVENTION
This invention relates to a novel oxepane isomer of 42-O-(2-hydroxy)ethyl-
rapamycin, a process for its preparation, and its use in treating, preventing or
inhibiting transplant rejection, graft vs. host disease, autoimmune diseases,
inflammatory diseases, adult T cell leukemia/lymphoma, solid tumors, fungal
infections, and hyperproliferative vascular disorders, among others.
The structure and synthesis of 40-O-(2-hydroxy)ethyl rapamycin, also known
as SDZ-RAD or RAD-666, is described in U.S. Patent No. 5,665,772 (Cottens, et al.)
and International Patent Publication No. WO 94/09010. When prepared according to
U.S. Patent No. 5,665,772, SDZ-RAD exists as a mixture containing about 95 wt% of
Isomer B and 30 wt% of Isomer C (the oxepane isomer).
40-O-(2-hydroxy)ethyl rapamycin is now known as 42-O-(2-hydroxy)ethyl
rapamycin, due to a change in numbering convention. SDZ-RAD is an analog of
rapamycin, which is a macrocyclic triene antibiotic produced naturally by
Streptomyces hygroscopicus.
Rapamycin has been found useful in an array of applications based on its
antitumoral and immunosuppressive effects. Uses include preventing or treating
systemic lupus erythematosis, pulmonary inflammation, insulin dependent diabetes
mellitus, smooth muscle cell proliferation and intimal thickening following vascular
surgery, adult T-cell leukemia/lymphoma, and ocular inflammation. Rapamycin and
rapamycin derivatives, including SDZ-RAD, continue to be studied for treatment of
these and other conditions.
SUMMARY OF THE INVENTION
The invention provides a purified oxepane isomer of 42-O-(2-hydroxy)ethyl
rapamycin (SDZ-RAD), known as SDZ-RAD Isomer C.
In another aspect, the invention provides a chemical process for preparing
purified SDZ-RAD Isomer C from SDZ-RAD.
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WO 2006/096325 PCT/US2006/006125
In another aspect, the invention provides for pharmaceutical compositions
containing purified SDZ-RAD Isomer C, for use in treating, inhibiting, and preventing
transplant rejection, graft vs. host disease, autoimmune diseases, inflammatory
diseases, adult T cell leukemia/lymphoma, solid tumors, fungal infections, and
hyperproliferative vascular disorders, among other diseases and disorders, in a
mammal in need thereof.
In another aspect, the invention provides for use of purified SDZ-RAD Isomer
C in preparing a medicament useful for treating, inhibiting, and preventing transplant
rejection, graft vs. host disease, autoimmune diseases, inflammatory diseases, adult T
cell leukemia/lymphoma, solid tumors, fungal infections, and hyperproliferative
vascular disorders, among other diseases and disorders, in a mammal in need thereof.
In yet another aspect, the invention provides a pharmaceutical kit or pack
containing a course of treatment for transplant rejection, graft vs. host disease,
autoimmune diseases, inflammatory diseases, adult T cell leukemia/lymphoma, solid
tumors, fungal infections, and hyperproliferative vascular disorders, among other
diseases and disorders, for an individual mammal, comprising a container having
purified SDZ-RAD Isomer C in unit dosage form.
DETAILED DESCRIPTION OF THE INVENTION
The invention provides a purified oxepane isomer of 42-O-(2-hydroxy)ethyl
rapamycin ("SDZ-RAD"), having the following structure:

which may also be described by the formula: 1,17-dihydroxy-l l-{2-[4-(2-hydroxy-
ethoxy)-3-methoxy-cyclohexyl]-lmethyl-ethyl}-18,29-dimethoxy-14,16,20,22,28,34-
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WO 2006/096325 PCT/US2006/006125
hexamethyl-10,36-dioxa-3-aza-tricyclo[29.4.1.0~3,8]hexatriaconta-15,23,25,27-
tetraen-2,9,13,19,35-pentaone.
As used here, the term "SDZ-RAD Isomer C" means a compound having the
preceding formula.
As used herein, the term "purified SDZ-RAD Isomer C" means a compound
having the preceding formula (i.e., SDZ-RAD Isomer C) and having a purity of
greater than 90 wt%, or greater than 95 wt%, 98 wt% or 99 wt%.
3
Also provided by the invention is a chemical process for preparing purified
SDZ-RAD Isomer C from SDZ-RAD. SDZ-RAD Isomer C exists in equilibrium with
SDZ-RAD Isomer B. Under conditions described herein, the equilibrium may be
driven from the greatly favored Isomer B state to the Isomer C state, as summarized in
Scheme I below. The preparation of SDZ-RAD (42-O-(2-hydroxy)ethyl rapamycin)
is described (under its previous name of 40-O-(2-hydroxy)ethyl rapamycin) in U.S.
Patent No. 5,665,772 (Cottens, et al). The conversion of SDZ-RAD Isomer B to
SDZ-RAD Isomer C through an intermediate may be accomplished in a mixture of
aqueous buffer and organic solvent in the range of about pH 4 to pH 10. In another
embodiment, the aqueous buffer has a pH of from about 4 to 10,5 to 9, 6 to 9, 7 to 9,
or 7.5 to 8.5. In a further embodiment, the aqueous buffer has a pH of about 8.5.


WO 2006/096325 PCT/US2OO6/006125
In one embodiment, the aqueous buffer is triethylammonium acetate (TEAA).
In other embodiments, any suitable aqueous buffer may be readily selected by one of
skill in the art, including, but not limited to, phosphate buffered saline, and water with
sodium citrate buffer. In one embodiment, the organic solvent is a polar aprotic
solvent, i.e., a solvent that has a molecular dipole but whose hydrogen atoms are not
bonded to oxygen or nitrogen, or a dipolar aprotic solvent, i.e., a solvent that has a
zero molecular dipole and whose hydrogen atoms are not bonded to oxygen or
nitrogen, but has charges on individual atoms. In one embodiment, the organic
solvent is acetonitrile (H-CO-N(CH3)2) or 1,4-dioxane (commonly referred to as
dioxane). In another embodiment, the organic solvent is 1,4-dioxane. In other
embodiments, the organic solvent is acetonitrile, dimethylsulfoxide (DMSO; CH3-SO-
CH3), dimethylformamide (HCON(CH3)2), an aldehyde or a ketone. In still other
embodiments, combinations of the above solvents are contemplated. In one
embodiment, the aqueous buffer and organic solvent are provided at a ratio of about
1:1.5 to about 1:1, by volume. However, other suitable buffer:solvent ratios will be
readily apparent to one of skill in the art. Similarly, other suitable aqueous buffers
and organic solvents useful in the process of the invention will be readily apparent to
those of skill in the art in view of the specification.
The conversion reaction from SDZ-RAD Isomer B to SDZ-RAD Isomer C
may be performed at room temperature, i.e., about 22°C to about 28°C. Alternatively,
the conversion may be performed at lower or higher temperatures, as needed.
Typically, conversion is allowed to proceed for about four (4) hours (or longer, as
needed or desired) and is stopped by extraction of SDZ-RAD isomer C with a suitable
organic solvent.
In one embodiment, the organic solvent used in the extraction is a polar
aprotic solvent. In one embodiment, the organic solvent used for extraction is
methylene chloride (CH2CI2). Other suitable solvents, or combinations thereof, will
be readily apparent to one of skill in the art in view of the specification.
Isolation of SDZ-RAD Isomer C may be accomplished using preparative
chromatography techniques, such as are well known to those of skill in the art. See
generally, Preparative Chromatography, by R.P.W. Scott, Chrom-Ed Book Series,
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available online, and Fundamentals of Preparative and Nonlinear Chromatography,
by Guiochon, G., et al., 1st Academic Press (1994).
SDZ-RAD Isomer C is useful as an immunosuppressive and anti-
inflammatory agent in treating, preventing or inhibiting, or in preparing medicaments
useful in the treatment, prevention, or inhibition of: transplant rejection, such as
kidney, heart, liver, lung, bone marrow, pancreas (islet cells), cornea, small bowel,
skin allografts and heart valve xenografts; graft vs. host disease; autoimmune
diseases, such as lupus, rheumatoid arthritis, diabetes mellitus, myasthenia gravis and
multiple sclerosis; inflammatory diseases, such as psoriasis, dermatitis, eczema,
sebormea, inflammatory bowel disease, pulmonary inflammation (including asthma,
chronic obstructive pulmonary disease, emphysema, acute respiratory distress
syndrome, bronchitis, and the like), and eye uveitis.
SDZ-RAD Isomer C also has antiturnor, antifungal, and antiproliferative
activities. Accordingly, SDZ-RAD Isomer C is useful in treating, preventing or
inhibiting, or in preparing medicaments useful in the treatment, prevention, or
inhibition of: solid tumors, including fibroids (uterine leiomyoma), sarcomas and
carcinomas, such as astrocytomas, prostate cancer, breast cancer, colon cancer, small
cell lung cancer, and ovarian cancer; anemia; adult T cell leukemia/lymphoma; mantle
cell lymphoma; fungal infections; and hyperproliferative vascular diseases such as
restenosis, graft vascular atherosclerosis, cardiovascular disease, cerebral vascular
disease, and peripheral vascular disease, such as coronary artery disease,
cereberovascular disease, arteriosclerosis, atherosclerosis, nonatheromatous
arteriosclerosis, vascular wall damage from cellular events leading toward immune-
mediated vascular damage, smooth muscle cell proliferation and intimal thickening
following vascular injury, and stroke or multiinfarct dementia. In one embodiment,
SDZ-RAD Isomer C is used to treat, prevent or inhibit restenosis following an
angioplasty procedure. When used for this purpose, SDZ-Isomer C may be
administered prior to, during, or subsequent to the procedure, or any combination of •
the above.
When adrninistered for the treatment or inhibition or prevention of any of the
above diseases or conditions, SDZ-RAD Isomer C may be administered to a mammal
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orally, parenterally, intravenously, intraperitoneally, intramuscularly, intranasally,
intrabronchially, transdermally, topically, intravaginally, or rectally.
Dosage requirements vary according to the route of administration. In one
embodiment, daily dosage of SDZ-RAD Isomer C is from approximately 0.1 mg/kg -
100 mg/kg, 0.001 mg/kg - 25 mg/kg, or approximately 0.01 mg/kg - 5 mg/kg. In other
embodiments, the dosage will vary depending on the route of administration, the
severity of symptoms and the subject being treated (i.e., patient history). Treatment
will generally be initiated with small dosages, i.e., dosages less than the optimum
dose. Thereafter, the dosage is increased until the optimum effect under the
circumstances is reached. Precise dosages may be determined by the administering
physician based on general experience and experience with the individual subject or
patient to be treated.
Also provided by the invention are pharmaceutical compositions of SDZ-RAD
Isomer C according to the invention and one or more pharmaceutically acceptable
carriers or excipients. The pharmaceutical carriers or excipients may be solid or
liquid. A solid carrier may include one or more substances which may also act as
flavoring agents, lubricants, solubilizers, suspending agents, chelating agents,
surfactants, fillers, glidants, compression aids, binders, tablet-disintegrating agents, or
encapsulating material. In powders, the carrier is a finely divided solid which is in
admixture with the finely divided active ingredient, i.e., SDZ-RAD Isomer C. In
tablets, the SDZ-RAD Isomer C is mixed with a carrier having the necessary
compression properties in suitable proportions and compacted in the shape and size
desired. In one embodiment, the powders and tablets contain about 99% or more of
the active ingredient. Suitable solid carriers include, for example, calcium phosphate,
magnesium stearate, talc, sugars, lactose, dextrin, starch, gelatin, cellulose, methyl
cellulose, sodium carboxymethyl cellulose, polyvinylpyrrolidine, low melting point
waxes and ion exchange resins.
Liquid carriers are used in preparing solutions, suspensions, emulsions,
syrups, elixers and pressurized compositions. The active ingredient may be dissolved
or suspended in a pharmaceutically acceptable liquid carrier such as water, an organic
solvent, a mixture of both, or pharmaceutically acceptable oils or fats. The liquid
carrier may contain other suitable pharmaceutical additives such as solubilizers,
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emulsifiers, buffers, preservatives, sweeteners, flavoring agents, suspending agents,
chelating agents, surfactants (e.g., polysorbate 20 or polysorbate 80), thickening
agents, colors, viscosity regulators, antioxidants, stabilizers or osmo-regulators.
Suitable liquid carriers for oral and parenteral administration include water
(containing additives as above, e.g., cellulose derivatives, such as sodium
carboxymethyl cellulose solution), alcohols (including monohydric alcohols and
polyhydric alcohols, e.g., glycols) and their derivatives, lecithins, and oils (e.g.,
fractionated coconut oil and arachis oil). For parenteral administration, the carrier
may also be an oily ester such as ethyl oleate and isopropyl myristate. Sterile liquid
carriers are useful in sterile liquid form compositions for parenteral administration.
The liquid carrier for pressurized compositions may be halogenated hydrocarbon or
other pharmaceutically acceptable propellant.
Oral formulations of SDZ-RAD Isomer C may include any conventionally
used oral forms, including tablets, capsules, buccal forms, troches, lozenges and oral
liquids, suspensions or solutions. Capsules may contain mixtures of SDZ-RAD
Isomer C with inert fillers and/or diluents such as the pharmaceutically acceptable
starches (e.g., corn, potato or tapioca starch), sugars, artificial sweetening agents,
powdered celluloses, such as crystalline and microcrystalline celluloses, flours,
gelatins, gums, etc.
Useful tablet formulations may be made by conventional compression, wet
granulation or dry granulation methods, and utilize pharmaceutically acceptable
diluents, binding agents, lubricants, disintegrants, surface modifying agents (including
surfactants), suspending or stabilizing agents, including but not limited to, magnesium
stearate, stearic acid, talc, sodium lauryl sulfate, rnicrocrystalline cellulose,
carboxymethylcellulose calcium, polyvinylpyrrolidone, gelatin, alginic acid, acacia
gum, xanthan gum, sodium citrate, complex silicates, calcium carbonate, glycine,
dextrin, sucrose, sorbitol, dicalcium phosphate, calcium sulfate, lactose, kaolin,
mannitol, sodium chloride, dry starches and powdered sugar. Surface modifying
agents useful in tablet formulations include nonionic and anionic surface modifying
agents. Representative examples of surface modifying agents include, but are not
limited to, poloxamer 188, benzalkonium chloride, calcium stearate, cetostearyl
alcohol, cetomacrogol emulsifying wax, sorbitan esters, colloidal silicon dioxide,
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phosphates, sodium dodecylsulfate, magnesium aluminum silicate, and
triethanolamine. Oral formulations herein may utilize standard delay or time release
formulations to alter the absorption of SDZ-RAD Isomer C. The oral formulation
may also include administering the active ingredient in water or fruit juice, containing
appropriate solubilizers or emulsifiers as needed.
The pharmaceutical forms of SDZ-RAD Isomer C suitable for injectable use
include sterile aqueous solutions or dispersions and sterile powders for the
extemporaneous preparation of sterile injectable solutions or dispersions. In all cases,
the form must be sterile and must be fluid to the extent that easy syringability exists.
It must be stable under the conditions of manufacture and storage and must be
preserved against the contaminating action of microorganisms such as bacteria and
fungi. The carrier may be a solvent or dispersion medium containing, for example,
water, ethanol, polyol (e.g., glycerol, propylene glycol and liquid polyethylene
glycol), suitable mixtures thereof, and vegetable oils.
The parenteral formulations useful in this invention may be used to produce a
dosage form that is suitable for administration by either direct injection or by addition
to sterile infusion fluids for intravenous infusion.
In another embodiment, SDZ-RAD Isomer C may be prepared for direct
administration to the airways in the form of an aerosol.
Transdermal administration may be accomplished through the use of a
transdermal patch containing the active compound and a carrier that is inert to the
active compound, is non-toxic to the skin, and allows delivery of the agent for
systemic absorption into the blood stream via the skin. Transdermal administration is
understood to include all administration across the surface of the body and the inner
linings of bodily passages including epithelial and mucosal tissues. Such
administrations may be carried out using SDZ-RAD Isomer C, in lotions, creams,
foams, patches, suspensions, solutions, and suppositories (rectal and vaginal).
The carrier may take any number of forms such as creams and ointments,
pastes, gels, and occlusive devices. The creams and ointments may be viscous liquid
or semisolid emulsions of either the oil-in-water or water-in-oil type. Pastes
comprised of absorptive powders dispersed in petroleum or hydrophilic petroleum
containing the active ingredient may also be suitable. A variety of occlusive devices
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may be used to release the active ingredient into the blood stream such as a semi-
permeable membrane covering a reservoir containing the active ingredient with or
without a carrier, or a matrix containing the active ingredient. Other occlusive
devices known in the literature are contemplated by the invention.
Suppository formulations may be made from materials known in formulation
art, including cocoa butter, with or without the addition of waxes to alter the
suppository's melting point, and glycerin. Water soluble suppository bases, such as
polyethylene glycols of various molecular weights, may also be used.
SDZ-RAD Isomer C may be formulated for any suitable delivery route and
vehicle and assembled in the form of a pharmaceutical pack or kit of parts.
Pharmaceutical packs or kits according to the invention are useful in the treatment,
inhibition, or prevention of any of the diseases or disorders listed herein, or in the
preparation of medicaments useful therefor. In one embodiment, the invention
includes a product containing SDZ-RAD Isomer C according to the invention for use
in treating a mammal, hi another embodiment, the invention includes a
pharmaceutical pack containing a course of treatment of a neoplasm for one
individual mammal, wherein the pack contains units of SDZ-RAD Isomer C in unit
dosage form. In still other embodiments, the above packs/kits include other
components, e.g., instructions for dilution, mixing and/or administration of SDZ-RAD
Isomer C, containers, syringes, needles, etc. Other such pack/kit components will be
readily apparent to one of skill in the art.
The reagents used in the preparation of SDZ-RAD Isomer C may be either
commercially obtained or may be prepared by standard procedures described in the
literature. The following examples are illustrative of the present invention, but are not
a limitation thereof.
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EXAMPLE 1
A. Materials and Instruments
SDZ-RAD was obtained from Chemical Development Wyeth-Ayerst
Research, Pearl River, NY. All solvents were HPLC grade and all other chemicals
were analytical reagent or equivalent. The preparative HPLC consisted of two
Dynamax™ solvent delivery systems (Model SD-1) and one Dynamax™ absorbance
detector (Model UV-1) from Rainin Instrument Company, Inc. (Woburn, MA). An
automatic speed-vac concentrator (Savant, Model AS 160) was obtained from Savant
Instruments, Inc. (Holbrook, NY) and a BUCHI rotary evaporation system (RE 260
and R 124) was obtained from Buchi (Flawil, Switzerland). 1H NMR and 13C data
were acquired on 600 MHz Varian Unity Plus spectrometers with a probe temperature
of 25°C. Mass spectra were obtained from API 365 mass spectrophotometer from PE
Sciex. All samples were prepared and run at ambient temperature.
B. Preparation of Compound
SDZ-RAD (40 mg; 0.042 mmol) was dissolved in 50 mL 40% 0.1 M TEAA
buffer pH 8.5 and 60% dioxane. The solution was stirred at room temperature
(approximately 22°C to 28°C) for 4 hours. The conversion was stopped by 2 x 50 mL
CH2CI2 extraction. The organic layer was reduced by rotary evaporation system to
dryness. The isolation of compound was performed by BDS-Hypersil-C18 column
(250 x 20 mm) using the mobile phase consisting of 60% dioxane, 40% water with
0.01M TEAA buffer pH 3.9. The flow rate was 12 mL/min. The fraction of isomer C
(28.9 min) was collected and extracted with CH2CI2 using a separatory funnel. The
organic layer was combined and washed with 2x50 mL water and then the organic
layer was dried with anhydrous Na2SO4. The organic solvent was reduced by rotary
evaporation to about 1 mL. The product was transferred into a vial and precipitated
by adding n-hexane. White powder was obtained by using N2 to blow away the
solvent and the vial was put into speed-vac to dry overnight. The purity of the isomer
C for each purification step was analyzed by HPLC. ESI mass spectrometry indicates
molecular ion [M+NR4]+ m/e 975.8, which is same as the reference sample of SDZ-
RAD.
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The NMR sample was prepared in acetone-d6. Table: Proton and Carbon
resonance assignment of SDZ-RAD isomer-C (NB # L20156-xxx) in Aceton-d6 at 25
°C,50OMHz(13C:100MHz)

c# δ13Cmajor δ1Hmajor δ13Cminor δ 1H minor 1HCorreltn HMBC
1 140.24 5.44 139.96 5.83 2,36
2 131.87 6.23 130.94 6.26 3,1
3 133.61 6.27 134.52 6.37 4,2
4 127.90* 6.49 128.13* 6.48 3,5
5 128.14 6.16 130.38 6.25 4,45
6 139.63 137.02
7 84.16 3.77 84.60 3.88 8 5,6,8,45,52
8 44.00 2.14;1.34 41.88 1.86;1.72 7,9 6,7
9 74.60 3.79 73.30 3.48 8,10 15
10 33.62 2.05eq1.52ax 36.01 1.78eq1.71ax 9,11
11 35.54 1.82eq1.24ax 35.28 1.75eq1.23ax 10,12
12 44.08* 3.24 44.15* 3.17 11 10,46
13 210.92 210.43
15 98.77 99.07
15(OH) 6.07 5.86 12,13,15,16
16 168.84 168.52
18 44.15 4.60eq2.93ax 39.87 4.16eq3.02ax 19 3.02/168.52
19 26.04 1.571.43 25.54 1.741.44 18,20
20 21.78* 1.661.51 21.69* 1.791.45 19,21
21 27.09 2.281.52 28.47 2.281.87 20,22
22 52.54 5.07 56.58 5.58 21
23 170.66 171.35
25 75.95 5.25 75.59 5.27 26,37 23,26,28,37,38,51
26 41.46 2.802.66 42.84 2.832.74 25
27 208.50 208.60
28 46.78 3.40 46.89 3.47 29,47 47
29 126.08 5.31 127.55 5.52 28,48 28,47
30 137.83 138.27
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PCT/US2006/006125

c# δ13Cmajor δ1Hmajor δ13Cminor δ lH minor lHCorreltn HMBC
31 77.63* 4.21 77.69* 4.22 29,32,48
31 (OH) 4.05 4.05 30,31,32
32 85.96 4.16 87.57 3.82 31,32,34
33 211.79 212.16
34 42.67 2.56 40.39 2.93 35,49
35 40.70 1.57(proR)1.16(proS) 41.99 1.55(proR)1.25(proS) 34,36
36 37.2 2.29 34.87 2.37 1,35,50
37 34.30 1.86 34.43 1.94 38,51
38 40.10 1.361.08 39.16 1.261.13 37,39
39 33.80* 1.36 33.96* 1.40 38,40,44
40 37.46 2.040.73 37.07 2.060.69 39,41 39,44
41 84.14* 3.12 84.27* 3.02 40,42
42 83.80* 3.11 83.75 3.11 41,43
43 31.24* 2.011.21 31.19* 2.011.21 42,44
44 32.38 1.680.93 32.03 1.720.91 39,43
45 10.74 1.72 10.99 1.67 5
46 17.17* 1.13 17.14* 1.14 12 11,12,13
47 15.96 0.95 17.02 0.09 28
48 14.56 1.83 13.42 1.75 29
49 14.32 0.93 14.94 1.02 34
50 22.04 1.03 21.43 1.06 36
51 15.63 0.87 14.56 0.93 37 37
52 56.51 3.12 56.25 3.18
53 58.04 3.27 58.35 3.26
54 57.69 3.27 58.35 3.26
55 72.37* 3.58* 72.40* 3.65*
56 62.58 3.58 62.58 3.58
EXAMPLE 2
Antifungal activity for the compound of this invention was established by
evaluation against several strains of fungi. Briefly, the following procedure was used
to evaluate such activity. A 96 well microtiter plate was filled (50 jaL/well) with
RPMI1640. The compounds to be evaluated were placed in appropriate wells and
serial diluted in successive wells to provide several dilutions. The concentrations
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ranged from 64 to 0.06 mg/mL. An adjusted inoculum of fungi (50 mL) was added to
each well and plates were incubated at 35°C for 24-48 hours. The minimum inhibitory
concentration (MIC) is the lowest concentration of compound that completely
inhibited growth of organism in the wells. The following table shows the results
obtained in this standard pharmacological test procedure. Where the same fungi name
is listed more than once, it indicates that more than one strain was evaluated. The
compound of this invention is listed in the table's heading as "Compound".
TABLE 1. Antifungal Activity (MIC in mg/mL)

Yeast/ Fungi ID Compound Nystatin Amphotericin B
Candida albicans 94-1 0.25 2 0.125
Candida albicans 1063 ≤0.06 1 ≤0.06
Candida albicans 1117 0.125 1 ≤0.06
Candida albicans ATCC 90028 0.125 1 0.125
Candida parapsilosis 94-9 0.125 1 0.125
Candida parapsilosis 94-8 0.125 2 ≤0.06
Candida parapsilosis ATCC 90018 0.25 2 ≤0.06
Candida pseudotropiadis ATCC 28838 5 0.06 1 ≤0.06
Candida tropicalis 94-14 0.125 2 0.125
Candida tropicalis 94-13 0.125 1 0.125
Candida krussii 94-2 0.5 1 0.125
Candida lusitmiae 94-3 ≤0.06 1 0.125
Candida zeylanoided 94-15 ≤0.06 1 0.25
Candida rugosa 94-10 0.25 1 0.25
Aspergillus fumigaius ATCC 26933 32 2 0.25
Aspergillus niger ATCC 16404 8 1 0.25
Aspergillus flavusr S506 8 2 0.50
The results obtained in this standard pharmacological test procedure
demonstrate the effectiveness of SDZ-RAD Isomer C.
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All patents, patent applications, articles, publications, and other documents
referenced herein are incorporated by reference. One of skill in the art will recognize
that minor modifications to the conditions and techniques in the embodiments
described herein may be varied without departing from the present invention as
defined by the following claims and are encompassed by the invention.
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CLAIMS:
1. An SDZ-RAD Isomer C compound of the structure

having a purity of greater than 90 wt%.
2. The SDZ-RAD Isomer C according to claim 1, wherein the purity of SDZ-
RAD Isomer C is greater than 95 wt%.
3. The SDZ-RAD Isomer C according to claim 1, wherein the purity of SDZ-
RAD Isomer C is greater than 99 wt%.
15
4. SDZ-RAD Isomer C compound of the structure


.WO 2006/096325 PCT/US2006/006125
(a) dissolving 42-O-(2-hydroxy)ethyl-raparnycin in a solution
containing an organic solvent and an aqueous buffer, said aqueous buffer having apH
of 4 to 10; and
(b) extracting the SDZ-RAD Isomer C into an organic solvent.

5. The SDZ-RAD Isomer C prepared according to claim 4, wherein the organic
solvent in step (a) is a polar aprotic or dipolar aprotic solvent.
6. The SDZ-RAD Isomer C prepared according to claim 5, wherein the organic
solvent in step (a) is acetonitrile or 1,4-dioxane.
7. The SDZ-RAD Isomer C prepared according to claim 6, wherein the organic
solvent is 1,4-dioxane.
8. The SDZ-RAD Isomer C prepared according to any one of claims 4 to 7,
wherein the aqueous buffer has a pH of about 7 to 9.
9. The SDZ-RAD Isomer C prepared according to claim 8, wherein the aqueous
buffer has apH of about 7.5 to 8.5.
10. The SDZ-RAD Isomer C prepared according to claim 9, wherein the aqueous
buffer has a pH of about 8.5.
11. The SDZ-RAD Isomer C prepared according to any one of claims 4 to 10,
wherein the organic solvent in step (b) is a polar aprotic solvent.
12. The SDZ-RAD Isomer C prepared according to claim 11, wherein the organic
solvent in step (b) is CH2CI2.
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for use as a medicament
18
17. SDZ-RAD Isomer C compound of the structure


(54) Title: OXEPANE ISOMBR OF 42-O-(2-HYDROXY)ETHYL-RAPAMYCIN
(57) Abstract: The invention provides a purified oxepane isomer of 42-0-(2-hydroxy)ethyl-rapamycin (SDZ-RAD Isomer C), a
chemical process for its preparation, as well as pharmaceutical compositions and packs containing SDZ-RAD Isomer C and methods
for its use as an immunosuppressive, anti-inflammatory, antifungal, antiproliferative and antitumor agent.
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The invention provides a purified oxepane isomer of 42-O-(2-hydroxy)ethyl-rapamycin (SDZ-RAD Isomer C), a
chemical process for its preparation, as well as pharmaceutical compositions and packs containing SDZ-RAD Isomer C and methods
for its use as an immunosuppressive, anti-inflammatory, antifungal, antiproliferative and antitumor agent.