This invention relates to the use of combinations of rapamycin
42-ester with 3-hydroxy-2-(hydroxymethyl)-2-methylpropionic acid (CCI-779) or 42-O-(2-hydroxy)ethyl
rapamycin and interferon-&agr;, for the preparation of medicaments for use in
antineoplastic combination chemotherapy
BACKGROUND OF THE INVENTION
Rapamycin is a macrocyclic triene antibiotic produced by
Streptomyces hygroscopicus, which was found to have antifungal activity,
particularly against Candida albicans, both in vitro and
in vivo [C. Vezina et al., J. Antibiot. 28, 721 (1975); S.N.
Sehgal et al., J. Antibiot. 28, 727 (1975); H. A. Baker
et al., J. Antibiot. 31, 539 (1978); U.S. Patent 3,929,992; and U.S.
Patent 3,993,749]. Additionally, rapamycin alone (U.S. Patent 4,885,171) or in combination
with picibanil (U.S. Patent 4,401,653) has been shown to have antitumor activity.
The immunosuppressive effects of rapamycin have been disclosed
in FASEB 3, 3411 (1989). Cyclosporin A and FK-506, other macrocyclic molecules,
also have been shown to be effective as immunosuppressive agents, therefore useful
in preventing transplant rejection [FASEB 3, 3411 (1989); FASEB
3, 5256 (1989); R. Y. Calne et al., Lancet 1183 (1978); and U.S. Patent
5,100,899]. R. Martel et al. [Can. J. Physiol. Pharmacol. 55,
48 (1977)] disclosed that rapamycin is effective in the experimental allergic
encephalomyelifis model, a model for multiple sclerosis; in the adjuvant arthritis
model, a model for rheumatoid arthritis; and effectively inhibited the formation
of IgE-like antibodies.
Rapamycin is also useful in preventing or treating systemic
lupus erythematosus [U.S. Patent 5,078,999], pulmonary inflammation [U.S. Patent
5,080,899], insulin dependent diabetes mellitus [U.S. Patent 5,321,009], skin disorders,
such as psoriasis [U.S. Patent 5,286,730], bowel disorders [U.S. Patent 5,286,731],
smooth muscle cell proliferation and intimal thickening following vascular injury
[U.S. Patents 5,288,711 and 5,516,781], adult T-cell leukemia/lymphoma [European
Patent Application 525,960 A1], ocular inflammation [U.S. Patent 5,387,589], malignant
carcinomas [U.S. Patent 5,206,018], cardiac inflammatory disease [U.S. Patent 5,496,832],
and anemia [U.S. Patent 5,561,138].
Rapamycin 42-ester with 3-hydroxy-2-(hydroxymethyl)-2-methylpropionic
acid (CCI-779) is an ester of rapamycin which has demonstrated significant inhibitory
effects on tumor growth in both in vitro and in vivo models. The preparation and
use of hydroxyesters of rapamycin, including CCI-779, are disclosed in U.S. Patents
5,362,718 and 6,277,983.
CCI-779 may delay the time to progression of tumors or
time to tumor recurrence which is more typical of cytostatic rather than cytotoxic
agents. CCI-779 is considered to have a mechanism of action that is similar to that
of sirolimus. CCI-779 binds to and forms a complex with the cytoplasmic protein
FKBP, which inhibits an enzyme, mTOR (mammalian target of rapamycin, also known
as FKBP12-rapamycin associated protein [FRAP]). Inhibition of mTOR's kinase activity
inhibits a variety of signal transduction pathways, including cytokine-stimulated
cell proliferation, translation of mRNAs for several key proteins that regulate
the G1 phase of the cell cycle, and IL-2-induced transcription, leading to inhibition
of progression of the cell cycle from G1 to S. The mechanism of action of CCI-779
that results in the G1-S phase block is novel for an anticancer drug.
In vitro, CCI-779 has been shown to inhibit the growth of a number of histologically
diverse tumor cells. Central nervous system (CNS) cancer, leukemia (T-cell), breast
cancer, prostate cancer, and melanoma lines were among the most sensitive to CCI-779.
The compound arrested cells in the G1 phase of the cell cycle.
In vivo studies in nude mice have demonstrated that CCI-779 has activity
against human tumor xenografts of diverse histological types. Gliomas were particularly
sensitive to CCI-779 and the compound was active in an orthotopic glioma model in
nude mice. Growth factor (platelet-derived)-induced stimulation of a human glioblastoma
cell line in vitro was markedly suppressed by CCI-779. The growth of several human
pancreatic tumors in nude mice as well as one of two breast cancer lines studied
in vivo also was inhibited by CCI-779.
Interferon-alfa is part of a family of naturally occurring
proteins, as well as a product produced by recombinant DNA techniques (including
interferon alfa-2a and interferon alfa-2b), that has been shown to have antiviral
and antitumor properties. While interferon-alfa produces immunomodulatory effects
and has antiangiogenic properties, its exact mechanism of action, at least in renal
cancer, is unknown. Indications include the treatment of patients with hairy cell
leukemia, chronic myelogenous leukemia, follicular lymphoma, cutaneous T cell lymphoma,
AIDS-related Kaposi's sarcoma, malignant melanoma, renal cancer, colorectal cancer,
other cancers (eg, cervical cancer, ovarian cancer), liver cirrhosis/liver cancer,
and the treatment of viral infections, including chronic hepatitis B, chronic hepatitis
C, and condylomata acuminata.
DESCRIPTION OF THE INVENTION
This invention provides the use of CCI-779.and interferon-alfa
in preparing a medicament for treating a neoplasm in a mammal in need thereof. In
particular, such a combination is useful in the treatment of renal cancer, soft
tissue cancer, breast cancer, neuroendocrine tumor of the lung, cervical cancer,
uterine cancer, head and neck cancer, glioma, non-small lung cell cancer, prostate
cancer, pancreatic cancer, lymphoma, melanoma, small cell lung cancer, ovarian cancer,
colon cancer, esophageal cancer, gastric cancer, leukemia, colorectal cancer, Kaposi's
sarcoma, liver cancer, and unknown primary cancer.
This invention also provides use of 42-O-(2-hydroxy)ethyl
rapamycin and interferon-alfa in preparing a medicament for treating a neoplasm
in a mammal in need thereof. The preparation of 42-O-(2-hydroxy)ethyl rapamycin
is described in U.S. Patent 5,665,772. The combination of CCI-779 or 42-O-(2-hydroxy)ethyl
rapamycin and interferon &agr; may also be used for preparing a medicament for
treating an estrogen receptor positive carcinoma in a mammal in need thereof such
as an estrogen receptor positive breast or ovarian cancer.
In the use aspect of this invention either the CCI-779
or the interferon &agr;, or both may be provided in subtherapeutically effective
amounts. Similarly either the 42-O-(2-hydroxy)ethyl rapamycin or the interferon
&agr; or both, may be provided in subtherapeutically effective amounts.
As used in accordance with this invention, the term "treatment"
means treating a mammal having a neoplastic disease by providing said mammal an
effective amount of a combination of CCI-779 and interferon-alfa with the purpose
of inhibiting growth of the neoplasm in such mammal, eradication of the neoplasm,
or palliation of the mammal.
As used in accordance with this invention, the term "providing,"
with respect to providing CCI-779 or 42-O-(2-hydroxy)ethyl rapamycin and interferon-alfa
combination (including simultaneous, separate or sequential administration of the
components of the combination), means directly administering CCI-779 along with
The preparation of CCI-779 is described in U.S. Patent
5,362,718. A regiospecific synthesis of CCI-779 is described in US Patent 6,277,983.
Interferon-alfa is commercially available as Roferon-A (interferon alfa-2a) and
Intron A (interferon alfa-2b).
The combinations of the invention may be in the form of
a kit of parts. The invention therefore includes a product containing (a) CCI-779
or 42-O-(2-hydroxy)ethyl rapamycin and (b) interferon (IFN) &agr; as a combined
preparation for simultaneous, separate or sequential use in treating a neoplasm
in a mammal in need thereof. The invention also includes a pharmaceutical pack containing
a course of treatment of a neoplasm for one individual mammal, wherein the pack
contains (a) units of CCI-779 or 42-O-(2-hydroxy)ethyl rapamycin in unit dosage
form and (b) units of IFN&agr; in unit dosage form.
The results examples illustrate the ability of an illustrative
combination of the invention, CCI-779 and interferon alfa, to treat a representative
carcinoma, renal cancer. The combination of the invention is useful in treating
soft tissue cancer, breast cancer, neuroendocrine tumor of the lung, cervical cancer,
uterine cancer, head and neck cancer, glioma, non-small lung cell cancer, prostate
cancer, pancreatic cancer, lymphoma, melanoma, small cell lung cancer, ovarian cancer,
colon cancer, esophageal cancer, gastric cancer, leukemia, colorectal cancer, Kaposi's
sarcoma, liver cancer, and unknown primary cancer.
As typical with chemotherapy, dosage regimens are closely
monitored by the treating physician, based on numerous factors including the severity
of the disease, response to the disease, any treatment related toxicities, age,
and health of the patient. Based on the results obtained with CCI-779, it is projected
that initial i.v. infusion dosages will be between about 0.1 and 100 mg/m2
when administered on a daily dosage regimen, and between about 1 and 1000 mg/m2
when administered on a weekly dosage regimen. Other dosage regimens and variations
are foreseeable, and will be determined through physician guidance. It is preferred
that CCI-779 is administered by i.v. infusion or orally, preferably in the form
of tablets or capsules. Other routes of administration are also feasible, such as
via implants, parenterally (besides i.v., such as intraperitoneal and subcutaneous
injections), rectally, intranasally, vaginally, and transdermally.
For interferon-alfa, it is projected that initial dosages
will be between about 100,000 and 20 million IU daily or between 500,000 and 75
million IU three times weekly. Other dosage regimens and variations are foreseeable,
and will be determined through physician guidance. It is preferred that interferon-alfa
is administered subcutaneously. Other routes of administration are also feasible,
such as intravenously or intramuscularly.
Dosage regimens are expected to vary according to the route
of administration. For example, dosages for oral administration are often up to
five to tenfold greater than for i.v. administration. The use of concomitant chemotherapeutic
agents often allows for dosage reduction of each particular agent, thereby increasing
the safety margin of the particular agents.
Oral formulations containing the active compounds of this
invention may comprise any conventionally used oral forms, including tablets, capsules,
buccal forms, troches, lozenges and oral liquids, suspensions or solutions. Capsules
may contain mixtures of the active compound(s) 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. 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 magnesium stearate, stearic acid, talc, sodium lauryl sulfate, microcrystalline
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, talc, dry starches and powdered sugar. Preferred
surface modifying agents include nonionic and anionic surface modifying agents.
Representative examples of surface modifying agents include poloxamer 188, benzalkonium
chloride, calcium stearate, cetostearyl alcohol, cetomacrogol emulsifying wax, sorbitan
esters, colloidal silicon dioxide, phosphates, sodium dodecylsulfate, magnesium
aluminum silicate, and triethanolamine. Oral formulations herein may utilize standard
delay or time release formulations to alter the absorption of the active compound(s).
The oral formulation may also consist of administering the active ingredient in
water or a fruit juice, containing appropriate solubilizers or emulsifiers as needed.
Particularly suitable oral formulations for rapamycin 42-ester
with 3-hydroxy-2-(hydroxymethyl)-2-methylpropionic acid are disclosed in USSN 60/411,264
and PCT/US03/29228. Such an oral formulation contains a granulation prepared using
a wet granulation process. The granulation contains CCI-779, a water soluble polymer,
a pH modifying agent, a surfactant, and an antioxidant. In one embodiment, the formulation
contains from 0.1 to 30%, from 0.5 to 25%, from 1 to 20%, from 5 to 15%, or from
7 to 12% (wt/wt) CCI-779, from 0.5 to 50%, from 1 to 40%, from 5 to 35%, from 10
to 25%, or from 15 to 20% (wt/wt) water soluble polymer, from 0.5 to 10%, 1 to 8%,
or 3 to 5% (wt/wt) surfactant, and from 0.001% to 1%, 0.01% to 1%, or 0.1% to 0.5%
The oral formulation may also contain suitable chelating
agents, fillers, binders, surfactants, and the like to facilitate the granulation
and tableting process. It is preferred that the wet granulation be performed with
a hydroalcoholic solvent system comprising water and an alcohol, with ethanol being
the preferred alcoholic component.
Typical water soluble polymers include polyvinylpyrrolidone
(PVP), hydroxypropylmethylcellulose (HPMC), polyethylene glycol (PEG), and cyclodextrin
or mixtures thereof. It is preferred that the water-soluble polymer is PVP, and
having a molecular weight of between 2.5 and 60 kilodaltons. Any given oral formulation
useful in the invention may contain multiple ingredients of each class of component.
For example, an oral formulation containing an antioxidant may contain one or more
antioxidants as the antioxidant component.
Acceptable pH modifying agents include citric acid, sodium
citrate, dilute HCl, and other mild acids or bases capable of buffering a solution
containing CCI-779 to a pH in the range of about 4 to about 6. Acceptable antioxidants
include, citric acid, d,l-&agr;-tocopherol, BHA, BHT, monothioglycerol, ascorbic
acid, and propyl gallate. It is expected that the antioxidants of the oral formulations
used in this invention will be used in concentrations ranging from 0.001% to 3%
wt/wt. Chelating agents, and other materials capable of binding metal ions, such
as ethylene diamine tetra acetic acid (EDTA) and its salts are capable of enhancing
the stability of CCI-779. Surfactants may include polysorbate 80, sodium lauryl
sulfate, sodium dodecyl sulfate, salts of bile acids (taurocholate, glycocholate,
cholate, deoxycholate) that may be combined with lecithin. Alternatively, ethoxylated
vegetable oils, such as Cremophor EL, vitamin E tocopherol propylene glycol succinate
(Vitamin E TGPS), polyoxyethylene-polyoxypropylene block copolymers, and poloxamers.
Binders, fillers, and disintegrants such as sucrose, lactose, microcrystalline cellulose,
croscarmellose sodium, magnesium stearate, gum acacia, cholesterol, tragacanth,
stearic acid, gelatin, casein, lecithin (phosphatides), carboxymethylcellulose calcium,
carboxymethylcellulose sodium, methylcellulose, hydroxyethylcellulose, hydroxypropylcellulose,
hydroxypropylmethycellulose phthalate, noncrystalline cellulose, cetostearyl alcohol,
cetyl alcohol, cetyl esters wax, dextrates, dextrin, lactose, dextrose, glyceryl
monooleate, glyceryl monostearate, glyceryl palmitostearate, polyoxyethylene alkyl
ethers, polyethylene glycols, polyoxyethylene castor oil derivatives, polyoxyethylene
stearates, and polyvinyl alcohol may also be incorporated into the oral formulation.
The oral formulation useful in the method of the invention
can be prepared by preparing an alcoholic solution comprising CCI-779 and an antioxidant,
and an aqueous solution comprising a water-soluble polymer, a surfactant, and a
pH modifier, in sufficient quantity to adjust the pH of the aqueous solution to
4 to 6. Suitable alcohols include methanol, ethanol, isopropanol, where ethanol
is the preferred alcohol. The solutions were mixed and added to a mixer containing
intragranular excipients. Alternatively, the alcoholic and aqueous solutions can
be added separately without mixing with each other. Such intragranular excipients
comprise binders and fillers to promote dissolution enhancement. Typical intragranular
excipients may include microcrystalline cellulose, lactose, and croscarmellose sodium.
The solid intragranular excipients are granulated with the solutions in the mixer
until a uniform granulation is achieved. The mixer can be a blender with intensifying
bar, a low shear granulator or a high shear granulator. The granulation is dried
in a fluid bed dryer at approximately 50°C, and milled using a suitable milling
device, such as a Fitz mill. The wet granulation and drying can be done in a fluid
bed granulator/dryer. The wet granulation can be dried using a tray drying oven.
If desired, the dried granulation can be further blended with extragranular fillers
and binders, such as microcrystalline cellulose, croscarmellose sodium, and magnesium
stearate in a blender, such as a V-blender, before compression into tablets.
Alternatively, some of the water-soluble polymer can be
contained in the intragranular excipients, and the aqueous and alcoholic solutions
added to the mixer containing the intragranular excipients stepwise. For example,
the order of addition to the mixer may be one half of the aqueous solution, followed
by the entire alcoholic solution, and then the remainder of the aqueous solution.
Other sequences of addition are possible and permissible in these solid oral formulations.
In some cases it may be desirable to administer the compounds
directly to the airways in the form of an aerosol.
The compounds may also be administered parenterally or
intraperitoneally. Solutions or suspensions of these active compounds as a free
base or pharmacologically acceptable salt can be prepared in water suitably mixed
with a surfactant such as hydroxy-propylcellulose. Dispersions can also be prepared
in glycerol, liquid polyethylene glycols and mixtures thereof in oils. Under ordinary
conditions of storage and use, these preparations contain a preservative to prevent
the growth of microorganisms.
The pharmaceutical forms 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 can 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
Particularly suitable injectable formulations for rapamycin
42-ester with 3-hydroxy-2-(hydroxymethyl)-2-methylpropionic acid are disclosed in
US Patent Application No. 10/626,943 and PCT/US03/223276. In this embodiment, the
injectable formulation useful in the invention provides a CCI-779 cosolvent concentrate
containing an parenterally acceptable solvent and an antioxidant as described above
and a parenteral formulation containing CCI-779, composed of CCI-779, an parenterally
acceptable cosolvent, an antioxidant, a diluent solvent, and a surfactant. Any given
formulation useful in this invention may contain multiple ingredients of each class
of component. For example, a parenterally acceptable solvent can include a non-alcoholic
solvent, an alcoholic solvent, or mixtures thereof. Examples of suitable non-alcoholic
solvents include, e.g., dimethylacetamide, dimethylsulfoxide or acetonitrile,
or mixtures thereof. "An alcoholic solvent," may contain one or more alcohols as
the alcoholic solvent component of the formulation. Examples of solvents useful
in the formulations invention include ethanol, propylene glycol, polyethylene glycol
300, polyethylene glycol 400, polyethylene glycol 600, polyethylene glycol 1000,
or mixtures thereof. These cosolvents are particularly desirable because degradation
via oxidation and lactone cleavage occurs to a lower extent for these cosolvents.
Further, ethanol and propylene glycol can be combined to produce a less flammable
product, but larger amounts of ethanol in the mixture generally result in better
chemical stability. A concentration of 30 to 100%v/v of ethanol in the mixture is
In this embodiment, the stability of CCI-779 in parenterally
acceptable alcoholic cosolvents is enhanced by addition of an antioxidant to the
formulation. Acceptable antioxidants include citric acid, d.l-&agr;-tocopherol,
BHA, BHT, monothioglycerol, ascorbic acid, propyl gallate, and mixtures thereof.
Generally, the parenteral formulations useful in this embodiment of the invention
will contain an antioxidant component(s) in a concentration ranging from 0.001%
to 1% w/v, or 0.01% to 0.5% w/v, of the cosolvent concentrate, although lower or
higher concentrations may be desired. Of the antioxidants, d,l-&agr;-tocopherol
is particularly desirable and is used at a concentration of 0.01 to 0.1% w/v with
a preferred concentration of 0.075% w/v of the cosolvent concentrate.
In certain embodiments, the antioxidant component of the
formulation of the invention also exhibits chelating activity. Examples of such
chelating agents include, e.g., citric acid, acetic acid, and ascorbic acid (which
may function as both a classic antioxidant and a chelating agent in the present
formulations). Other chelating agents include such materials as are capable of binding
metal ions in solution, such as ethylene diamine tetra acetic acid (EDTA), its salts,
or amino acids such as glycine are capable of enhancing the stability of CCI-779.
In some embodiments, components with chelating activity are included in the formulations
of the invention as the sole "antioxidant component". Typically, such metal-binding
components, when acting as chelating agents are used in the lower end of the range
of concentrations for the antioxidant component provided herein. In one example,
citric acid enhanced the stability of CCI-779 when used at a concentration of less
than 0.01% w/v. Higher concentrations are less stable solutions and thus, less desirable
for products to be subject to long-term storage in liquid form. Additionally, such
chelating agents may be used in combination with other antioxidants as part of the
antioxidant component of the invention. For example, an acceptable formulation may
contain both citric acid and d,l-&agr;-tocopherol. Optimal concentrations for
the selected antioxidant(s) can be readily determined by one of skill in the art,
based upon the information provided herein.
Advantageously, in certain embodiments of the parenteral
formulations useful in the invention, precipitation of CCI-779 upon dilution with
aqueous infusion solutions or blood is prevented through the use of a surfactant
contained in the diluent solution. The most important component of the diluent is
a parenterally acceptable surfactant. One particularly desirable surfactant is polysorbate
20 or polysorbate 80. However, one of skill in the art may readily select other
suitable surfactants from among salts of bile acids (taurocholate, glycocholate,
cholate, deoxycholate, etc.) which are optionally combined with lecithin. Alternatively,
ethoxylated vegetable oils, such as a pegylated castor oil [e.g., such as PEG-35
castor oil which is sold, e.g., under the name Cremophor EL, BASF], vitamin E tocopherol
propylene glycol succinate (Vitamin E TGPS), and polyoxyethylene-polyoxypropylene
block copolymers can be used in the diluent as a surfactant, as well as other members
of the polysorbate family such as polysorbate 20 or 60 Other components of the diluent
may include water, ethanol, polyethylene glycol 300, polyethylene 400, polyethylene
600, polyethylene 1000, or blends containing one or more of these polyethylene glycols,
propylene glycol and other parenterally acceptable cosolvents or agents to adjust
solution osmolarity such as sodium chloride, lactose, mannitol or other parenterally
acceptable sugars, polyols and electrolytes. It is expected that the surfactant
will comprise 2 to 100% w/v of the diluent solution, 5 to 80% w/v, 10 to 75% w/v,
15 to 60 % w/v, and preferably, at least 5% w/v, or at least 10% w/v, of the diluent
A parenteral formulation useful in the invention can be
prepared as a single solution, or preferably can be prepared as a cosolvent concentrate
containing CCI-779, an alcoholic solvent, and an antioxidant, which is subsequently
combined with a diluent that contains a diluent solvent and suitable surfactant.
Prior to use, the cosolvent concentrate is mixed with a diluent comprising a diluent
solvent, and a surfactant. When CCI-779 is prepared as a cosolvent concentrate according
to this invention, the concentrate can contain concentrations of CCI-779 from 0.05
mg/mL, from 2.5 mg/mL, from 5 mg/mL, from 10 mg/mL or from 25 mg/mL up to approximately
50 mg/ml. The concentrate can be mixed with the diluent up to approximately 1 part
concentrate to 1 part diluent, to give parenteral formulations having concentrations
of CCI-779 from 1mg/mL, from 5 mg/mL, from 10 mg/mL, from 20 mg/mL, up to approximately
25 mg/ml. For example the concentration of CCI-779 in the parenteral formulation
may be from about 2.5 to 10 mg/mL. This invention also covers the use of formulations
having lesser concentrations of CCI-779 in the cosolvent concentrate, and formulations
in which one part of the concentrate is mixed with greater than 1 part of the diluent,
e.g., concentrate: diluent in a ratio of about 1:1.5, 1:2, 1 :3, 1:4 ,1:5, or 1:9
v/v and so on, to CCI-779 parenteral formulations having a CCI-779 concentration
down to the lowest levels of detection.
Typically the antioxidant may comprise from about 0.0005
to 0.5% w/v of the formulation. The surfactant may for example comprise from about
0.5% to about 10% w/v of the formulation. The alcoholic solvent may for example
comprise from about 10% to about 90% w/v of the formulation.
The parenteral formulations useful in this invention can
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.
For the purposes of this disclosure, transdermal administrations
are understood to include all administrations 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 the present compounds, or pharmaceutically
acceptable salts thereof, in lotions, creams, foams, patches, suspensions, solutions,
and suppositories (rectal and vaginal).
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. 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 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 are known in the literature.
Suppository formulations may be made from traditional materials,
including cocoa butter, with or without the addition of waxes to after the suppository's
melting point, and glycerin. Water soluble suppository bases, such as polyethylene
glycols of various molecular weights, may also be used.
The following examples are illustrative of the present
Example 1 - CCI-779 and Interferon-&agr; Combination effective
against tumor cells
CCI-779 was evaluated in combination with Interferon-alpha
(IFN-&agr;) in the HTB-44 human mouse xenograft standard pharmacological test
procedure of renal cancer. The human renal cell line HTB-44 (also referred to in
the scientific literature as A498) is derived from a patient with a clear cell carcinoma
that had lost expression of the von Hippel-Lindau (VHL) gene. These types of tumors
are representative of the large majority (≈80%) of sporadic renal cell carcinomas.
CCI-779 was evaluated as a single agent on a weekly schedule
against large (>500mg) HTB-44 tumors in nude mouse xenografts. Dosing was from
10 mg/kg to 75 mg/kg intravenously once a week beginning on day 0 when tumors had
reached a size of about 500 mm3. All doses tested were similarly effective
(30-35 % inhibition of tumor growth) and a dose of 25 mg/kg was chosen to combine
Similarly, a dose response of IFN-&agr; as a single agent
was performed. Doses of 1 million units or 0.5 million units 3 times per week were
similarly effective, suggesting that this was the plateau range for maximally effective
treatment with IFN-&agr; (data not shown). Therefore, a dose of 1 million units
3 times per week intraperitoneally was chosen to combine with CCI-779 at 25 mg/kg
iv once per week. Groups of 10 mice were treated with CCI-779 alone, IFN-&agr;
alone, or the combination (Table 1). IFN-&agr; was given on days 1, 3, and 5 and
CCI-779 on day 6 of each week for 4 weeks. Dosing began after tumors had reached
a size of about 600 mg.
Table 1. Effect of combination therapy with CCI-779 and Interferon-&agr;
on HTB-44 human renal tumor growth in nude mice
Tumor mass (mg)
CCI-779 (25 mg/kg)
Interferon &agr; (1 x 106 units)
CCI-779 + IFN &agr;
Number in parenthesis = % of
Treatment with either CCI-779 alone or IFN-&agr; alone
resulted in retardation of tumor growth without tumor regression. When combined
with IFN-&agr;, CCI-779 induced a 36% regression in the size of HTB-44 renal cell
tumors growing in nude mice. As single agents, neither compound induced tumor regression
although both showed cytostatic activity. A three fold higher (75mg/kg) dose of
CCI-779 than that used in the combination study also did not induce tumor regression.
Higher doses of IFN-&agr; are also unlikely to induce regression since there was
no difference in single agent IFN at 0.5 or 1.0 million units suggesting that the
1.0 million units used in the combination study was in the plateau range of maximal
activity. Taken together these data show that CCl-779 and IFN-&agr; are synergistic
in this test procedure in that they were able to achieve an effect (tumor regression)
not attainable with single agent treatment.
Example 2 - CCI-779 and Interferon-&agr; combination active
In xenograft models of RCC such as shown in Example 1,
the combination resulted in tumor regression while each agent resulted only in tumor
growth inhibition. Thus, the combination of CCI-779 and IFN was evaluated in RCC
patients (pts) in a phase 1 study.
In an open-label, ascending-dose, single-arm study, CCI-779
was given IV once weekly, with IFN given subcutaneously 3 times weekly. IFN was
given alone the 1 st treatment week. The starting dose levels were 6 million units
(MU) IFN and 5 mg CCI-779. CCI-779 dose-escalation steps were 10 mg, 15 mg, and
25 mg. Once the maximum dose of CCI-779 is determined, the dose level of IFN can
be escalated to 9 MU (at a reduced CCI-779 dose, if necessary). Dose escalation
was based on a safety evaluation of pts (≥6/cohort) after 4 weeks of treatment.
In preliminary results, the number of pts at each dose
level was 5 mg: 7 pts, 10 mg: 6, 15 mg: 5, 25 mg: 2. Median age was 55 yrs (range,
40-72 yrs), ECOG performance status 0: 45%, 1: 55%. [ECOG performance status refers
to criteria established by the Eastern Cooperative Oncology Group (ECOG) and published,
e.g., Oken, M.M., et al., Toxicity And Response Criteria Of The Eastern Cooperative
Oncology Group. Am J Clin Oncol 5:649-655, 1982]. Prior treatment
with IL-2: 55%. Of 20 pts, 15 have been on study for 7.6+ mos. CCI-779-related adverse
events (AEs) with an overall frequency of ≥20% (n=18) included mucositis
(44%), nausea (39%), asthenia (39%), anemia (33%), anorexia (33%), hyperlipidemia
(28%), diarrhea (28%), leukopenia (22%), chills (22%), fever (22%), allergic reaction
(22%), taste perversion (22%). Gr 3-4 CCI-779-related AEs in ≥2 pts were
hyperlipidemia (4), leukopenia (3), hyperglycemia (2). AE-related dose reductions
or delays occurred in 7 pts. No drug-related deaths occurred. Partial responses
were reported for 2 pts, 5 pts had stable disease, 5 had progressive disease, the
remainder were too early to evaluate.
Up to 40 patients are being evaluated at the maximum tolerated
dose (MTD). In summary, 71 pts with advanced RCC were enrolled; 27 continue treatment.
Patients (73% men, 27% women) had ECOG performance status of 0: 53% and 1: 46% and
median age of 59 yrs (range, 35-80); 45% had prior IL-2 treatment. In dose escalation,
patients received 6 MU IFN&agr; with CCI-779 at 5 mg (7 pts), 10 mg (6), 15 mg
(6), 20 mg (6), or 25 mg (7); 6 pts also received 9 MU IFN&agr; with 15 mg CCI-779.
Based on dose-limiting toxicities, 15 mg CCI-779, 6 MU IFN&agr; was selected as
the MTD. To date, 33 additional patients have been accrued at the MTD. Grade 3-4
CCI-779-related adverse events with overall frequency ≥ 5% (n=53) were leukopenia
(25%), hyperlipidemia (15%), asthenia (13%), AST increase (8%), mucositis (6%),
anemia (6%), thrombocytopenia (6%), and rash (6%). Four patients were removed from
study due to CCI-779-related toxicity. Approximately 50% of MTD patients have required
CCI-779 dose reductions in subsequent cycles. Median time on treatment for all cohorts
was 7 mo, 36 have continued for ≥ 6 mo of whom 9 have continued for >
12 mo. Preliminary tumor responses (RECIST) in 55 pts were confirmed partial response,
7 pts (13%); stable disease, 39 (71%, 19 pts ≥ 6 mo); and progressive disease,
Combination therapy of CCI-779 and IFN has been generally
well tolerated in pts with advanced RCC and antitumor activity was observed.
Example 3 - Tablets each containing 2.5 mg of interferon
&agr; and also tablets each containing a dose of CCI-779 as mentioned in Example
1 are packaged in a container to provide a course of treatment for a patient.