EP0828508A2 - Compositions of interleukin and pyrimidine nucleosides - Google Patents
Compositions of interleukin and pyrimidine nucleosidesInfo
- Publication number
- EP0828508A2 EP0828508A2 EP96917379A EP96917379A EP0828508A2 EP 0828508 A2 EP0828508 A2 EP 0828508A2 EP 96917379 A EP96917379 A EP 96917379A EP 96917379 A EP96917379 A EP 96917379A EP 0828508 A2 EP0828508 A2 EP 0828508A2
- Authority
- EP
- European Patent Office
- Prior art keywords
- deoxy
- cytidine
- fluoro
- fluorocytidine
- pyrimidine nucleoside
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/19—Cytokines; Lymphokines; Interferons
- A61K38/20—Interleukins [IL]
- A61K38/208—IL-12
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
Definitions
- This invention is concerned with a novel pharmaceutical composition. More particularly, this invention is concerned with a synergistic antitumor pharmaceutical composition comprising an effective amount of interleukin- 12 (IL-12) and a pyrimidine nucleoside, as well as a hydrate or solvate thereof, that is converted into fluorouracil or its derivative, and pharma ⁇ ceutically acceptable carrier, a synergistic antitumor pharmaceutical composition for the treatment of various cancer and a method of treating various cancers.
- IL-12 interleukin- 12
- a pyrimidine nucleoside as well as a hydrate or solvate thereof, that is converted into fluorouracil or its derivative
- pharma ⁇ ceutically acceptable carrier a synergistic antitumor pharmaceutical composition for the treatment of various cancer and a method of treating various cancers.
- doxifluridine 5'-Deoxy-5-fluorouridine
- doxifluridine a pyrimidine nucleoside
- Doxifluridine is converted into the active drug 5-FU by pyrimidine nucleoside phosphorylases (PyNPase) in vivo, both thymidine and uridine phosphorylases. Therefore, PyNPase is essential for the efficacy of doxifluridine. In fact, tumors with very low levels of this enzyme were refractory to doxifluridine, and PyNPase gene transfection made the tumors more susceptible to this drug.
- IL-12 up-regulates PyNPase activity in tumor tissues and consequently enhances the antitumor activity of doxifluridine.
- IL-12 also enhanced the activity of 5'-deoxy-5-fluoro-N4-(n- pentylcarbonyl)cytidine (capecitabine), which generates doxifluridine and is then converted to 5-FU by PyNPase.
- IL-12 enhanced the anti ⁇ tumor activity of 5-FU to a lesser extent than the anti-tumor activity of doxifluridine.
- cytokines such as IL-la, TNF-a and IFN-g up-regulate PyNPase activity in tumor cell cultures and consequently enhance the susceptibility to doxifluridine (cf. Eda et al. Cancer Chemother Pharmacol. (1993) 32:333-339, and Jpn. J. Cancer Res. 84, 341- 347, March 1993).
- cytokines when given parenterally are distributed to various normal tissues through the circulation and cause systemic side effects, such as flu-like syndrome, leukopenia, hypotension, etc.
- these cytokines distributed to normal tissues as well as tumor tissues would enhance PyNPase activity there and make both the normal and tumor tissues more susceptible to doxifluridine. Therefore, these cytokines would enhance both the efficacy and toxicity of doxifluridine when given in combination.
- IL-12 given parenterally however, induced much higher levels of IFN-g in tumor tissues than in normal tissues. Therefore, IL-12 given parenterally enhances PyNPase activity preferentially in tumor tissues without causing IFN-g-associated systemic side effects.
- the pyrimidine nucleoside is an uridine, cytidine or its derivative represented by the following formula (I) or (II), respectively
- R ⁇ is hydrogen or an radical which is easily hydrolyzable under physiological conditions
- R-2 is hydrogen, cyano, fluorine,lower alkyl or lower alkylidene which may be substituted with one or two fluorine atom(s), or O l
- B ⁇ is lower alkyl, h droxymethyl, or CH2OR 1 , as well as a hydrate or solvate thereof.
- Preferred radicals which are easily hydrolyzable under physiological conditions of Rl in the above formulae (I) and (II) are R ⁇ CO-, R ⁇ OCO- or R SCO- wherein R 4 is alkyl, cycloalkyl, aralkyl or aryl.
- preferred alkyl, cycloalkyl, aralkyl or aryl radical represented by R 4 are a saturated, straight or branched hydrocarbon radical [wherein the number of carbon atoms in the longest straight chain of this hydrocarbon radical ranges from three to seven], or a radical of the formula (CH2)n-Y[ n which n is an integer from 0 to 4, when Y is cyclohexyl, or n is an integer from 2 to 4, when Y is lower alkoxy having 1 to 4 carbon atom(s) or phenyl] .
- a saturated, straight or branched hydrocarbon radical [wherein the number of carbon atoms in the longest straight chain of this hydrocarbon radical ranges from three to seven]" preferably signifies n-propyl, l-isopropyl-2-methylpropyl, 1,1,2-trimethylpropyl, n-butyl, isobutyl, 2-ethylbutyl, 3,3-dimethylbutyl, n-pentyl, isopentyl, neopentyl, 2-propylpentyl, n-hexyl, 2-ethylhexyl, n-heptyl, allyl, 2-buten-l-yl, 3-buten-l-yl, 3-penten-l-yl, 4-penten-l-yl, 3-hexen-l-yl, 4-hexen-l-yl, 5 hexen-1-yl, and the like.
- a radical of the formula (CE_2)n-Y [in which n is an integer from 0 to 4, when Y is a cyclohexyl radical, or n is an integer from 2 to 4, when Y is a lower alkoxy radical having from 1 to 4 carbon atom(s) or a phenyl radical]" preferably siginifies cyclohexyl, cyclohexylmethyl,
- Preferred pyrimidine nucleoside for the present invention are:
- IL-12 is a heterodimeric cytokine which is produced by antigen presenting cells and serves as a pivotal regulator of T and NK cell function (cf. Stern, A.S. et al. Proc. Natl. Acad. Sci. USA. (1990) 87, 6808-6812 and Kobayashi, M. et al. J. Exp. Med. (1989) 170, 827-845).
- Biological activities associated with IL-12 include its ability to enhance the lytic activity of natural killer/lymphokine activated killer cells, to induce the secretion of interferon-g (IFN-g) by both resting and activated T and NK cells, to stimulate the proliferation of activated T and NK cells, to facilitate cytotoxic T lymphocyte responses and to play a critical and unique role in promoting Th-1 type cytokine responses, thereby facilitating cell-mediated immunity (cf. Brunda, M. J. J. Leukocyte Biol. (1994) 55, 280-288 and Taniguchi, G. Blood (1994) 84, 4008-4027).
- IFN-g interferon-g
- IL-12 both human type and murine type, is composed of two disulfide- bonded glycoprotein subunits approximately 35 KDa and 40 KDa in size.
- cDNAs encoding each subunit of IL-12 have been cloned and coexpressed in Chinese Hamster Ovary (CHO) cells to yield the secreted, bioactive, heterodimeric lymphokine (Gubler, U. et al. Proc. Natl. Acad. Sci. USA. (1991) 88, 4143-4147 and Schaenhaut, D.S. et al. J. Immunol. (1992) 148, 3433- 3440)
- a clone of transfected CHO cells secreting recombinant IL-12 was selected.
- Recombinant IL-12 was purified from the culture supernatant of CHO cells grown in a serum-free medium, by ion exchange and gel filtration chromatography.
- the pharmaceutical compostion of the present invention can be administered in any form, for example, tablets, pills, suppositories, capsules, granules, powders, etc. or emulsions.
- the pharmaceutical composition of the present invention are especially suitable for intramuscular, subcutaneous, or intravenous administration.
- Pharmaceutically acceptable carriers and excipients useful in formulating the pharmaceutical composition of the present invention are those commonly used.
- Pharmaceutically acceptable materials can be an organic or inorganic inert carrier material suitable for enteral, percutaneous or parenteral administration such as water, gelatine, gum arabic, lactose, starch, magnesium stearate, talc, vegetable oils, polyalkylene glycols and petroleum jelly.
- the pharmaceutical composition provided by the present invention can be administered orally, e.g.
- the administration can also be carried out parenterally, e.g. in form of sterile solutions, suspensions or emulsions; or locally, e.g. in form of solutions, suspensions, salves, powders or aerosols.
- the pharmaceutical composition can be sterilized and/or can contain further adjuvants such as preserving, stabilizing, setting, emulsifying agents, flavor-improving, salts for variation of the osmotic pressure or substances acting as buffers.
- the synergistic antitumor pharmaceutical composition of the present invention comprises a single pharmaceutical composition as well as a kit of pharmaceutical compositions each containing the individual active ingredient in a desirable dosage form, thus, the present invention is also concerned with a kit for the treatment of colorectal cancer, breast cancer, stomach cancer, lung cancer, cervical cancer, bladder cancer and other malignant diseases which comprises as a first pharmaceutical composition containing an effective amount of IL-12 and a pharmaceutically acceptable carrier, and as a second pharmaceutical composition containing an effective amount of pyrimidine nucleoside derivative, as well as hydrate or solvate thereof and a pharmaceutically acceptable carrier.
- Dosage ranges for the pharmaceutical composition of the present invention can easily be determined by one skilled in the art, and depend on the route of administration, the age, weight and condition of the patient and the particular disease to be treated.
- an approximate range from about 0.05 mg/body/day to about 500 mg/body/day of IL-12, and about 50 mg/body/day to about 20,000 mg/body/day of pyrimidine nucleoside generally range from about 1:100 to about 1:400,000.
- a weight ratio from about 1:1,000 to about 1:10,000 is preferred.
- Rectal administration and intravenous injection are preferred routes of administration of the pharmaceutical composition according to the present invention.
- compositions of the present invention are useful for the treatment of colorectal cancer, breast cancer, stomach cancer, lung cancer, cervicial cancer, bladder cancer, and other malignant diseases and the like.
- mouse IL-12 (mIL-12) at 0.1 mg/mouse or vehicle (0.1 mg/ml of mouse serum albumin dissolved in phosphate-buffered saline) daily for 7 days starting at day 7 and day 8, respectively, after the tumor inoculation.
- mIL-12 mouse IL-12
- vehicle 0.1 mg/ml of mouse serum albumin dissolved in phosphate-buffered saline
- PyNPase activity in the tumor tissues was measured as described in Eda et al. Cancer Chemother. Pharmacol. 1993; 32:333.
- Mouse IFN-g (m IFN-g) levels in the tumor tissue were also measured by a commercially available ELISA system (Intertest g, Genzyme).
- mIL-12 enhanced PyNPase activity 11.9 fold in A75 ⁇ tumors and 2.4 fold in Meth A tumors. This is probably the result of the up- regulation of mIFN-g, which is an up-regulator of PyNPase.
- mice were inoculated with A7 ⁇ mammary adenocarcinoma (2 x 10 5 cells). The mice were given s.c. mIL-12 at 0.1 mg/mouse or vehicle (0.1 mg/ml of mouse serum albumin dissolved in phosphate-buffered saline) daily for 7 days starting at day 6 after the tumor inoculation. One day thereafter, mIFN-g levels in the serum and tissue homogenates of tumor and other organs were measured by ELISA system as mentioned above.
- mIL-12 greatly increased mIFN-g levels in tumors as compared with those in normal organs.
- the tumor tissue level of mIFN-g is 3 to 7 fold higher than those of normal tissues so far examined and 50 fold higher than those in the circulation.
- A7 ⁇ (2 x 10 5 cells) was inoculated s.c. into female C ⁇ 7BL/6 mice.
- the mice were given mIL-12 (0.03 ⁇ g/mouse, s.c), doxifluridine (1.5 mmol/kg, p.o.) and their combination, daily for 4 weeks, starting from day 9 after the tumor inoculation.
- Doxifluridine as a single agent did not show activity in tumor growth inhibition because A75 ⁇ tumor has only low levels of PyNPase, whereas mIL-12 was effective (Table 3). mIL-12 in combination with doxifluridine was much more effective than mIL-12 alone and regressed the tumor.
- Control 28 (23 - 42) 0 o/ ⁇ doxifluridine 29 (22 - 36) 4 0/5
- Meth A fibrosarcoma (5 x 10 5 cells) was inoculated s.c. into male BALB/c mice.
- the mice were given doxifluridine (O. ⁇ mmol/kg, p.o.), capecitabine (1.0 mmol/kg, p.o.), ⁇ -fluorouracil (0.07 ⁇ mmol kg, p.o.), mIL-12 (0.03 ⁇ g/mouse, s.c), and their combination, daily for 3 weeks, starting from day 8 after the tumor inoculation.
- Example 1 illustrate a pharmaceutical preparation of the present invention and do not limit the scope of the present invention.
- a kit having the following components A and B for treatment of colorectal cancer was manufactured in a conventional manner: Component A
Abstract
A synergistic antitumor pharmaceutical composition comprising an effective amount of interleukin-12 and a pyrimidine nucleoside derivative as well as a hydrate or solvate thereof that is converted into fluorouracil or its derivative, and a pharmaceutically acceptable carrier.
Description
Compositions of interleukin and pyrimidine nucleosides
This invention is concerned with a novel pharmaceutical composition. More particularly, this invention is concerned with a synergistic antitumor pharmaceutical composition comprising an effective amount of interleukin- 12 (IL-12) and a pyrimidine nucleoside, as well as a hydrate or solvate thereof, that is converted into fluorouracil or its derivative, and pharma¬ ceutically acceptable carrier, a synergistic antitumor pharmaceutical composition for the treatment of various cancer and a method of treating various cancers.
5'-Deoxy-5-fluorouridine (doxifluridine), a pyrimidine nucleoside, is effective in the treatment of various malignant diseases. Doxifluridine is converted into the active drug 5-FU by pyrimidine nucleoside phosphorylases (PyNPase) in vivo, both thymidine and uridine phosphorylases. Therefore, PyNPase is essential for the efficacy of doxifluridine. In fact, tumors with very low levels of this enzyme were refractory to doxifluridine, and PyNPase gene transfection made the tumors more susceptible to this drug. Now, it has surprisingly been found that IL-12 up-regulates PyNPase activity in tumor tissues and consequently enhances the antitumor activity of doxifluridine. IL-12 also enhanced the activity of 5'-deoxy-5-fluoro-N4-(n- pentylcarbonyl)cytidine (capecitabine), which generates doxifluridine and is then converted to 5-FU by PyNPase. In contrast, IL-12 enhanced the anti¬ tumor activity of 5-FU to a lesser extent than the anti-tumor activity of doxifluridine.
It has been reported that some inflammatory cytokines, such as IL-la, TNF-a and IFN-g up-regulate PyNPase activity in tumor cell cultures and consequently enhance the susceptibility to doxifluridine (cf. Eda et al. Cancer Chemother Pharmacol. (1993) 32:333-339, and Jpn. J. Cancer Res. 84, 341- 347, March 1993). These cytokines when given parenterally are distributed to various normal tissues through the circulation and cause systemic side effects, such as flu-like syndrome, leukopenia, hypotension, etc. In addition,
these cytokines distributed to normal tissues as well as tumor tissues would enhance PyNPase activity there and make both the normal and tumor tissues more susceptible to doxifluridine. Therefore, these cytokines would enhance both the efficacy and toxicity of doxifluridine when given in combination. IL-12 given parenterally, however, induced much higher levels of IFN-g in tumor tissues than in normal tissues. Therefore, IL-12 given parenterally enhances PyNPase activity preferentially in tumor tissues without causing IFN-g-associated systemic side effects.
In a preferred embodiment of the present invention, the pyrimidine nucleoside is an uridine, cytidine or its derivative represented by the following formula (I) or (II), respectively
wherein R^ is hydrogen or an radical which is easily hydrolyzable under physiological conditions; R-2 is hydrogen, cyano, fluorine,lower alkyl or lower alkylidene which may be substituted with one or two fluorine atom(s), or O l; and Bβ is lower alkyl, h droxymethyl, or CH2OR1, as well as a hydrate or solvate thereof.
Preferred radicals which are easily hydrolyzable under physiological conditions of Rl in the above formulae (I) and (II) are R^CO-, R^OCO- or R SCO- wherein R4 is alkyl, cycloalkyl, aralkyl or aryl.
Furthermore, preferred alkyl, cycloalkyl, aralkyl or aryl radical represented by R4 are a saturated, straight or branched hydrocarbon radical [wherein the number of carbon atoms in the longest straight chain of this hydrocarbon radical ranges from three to seven], or a radical of the formula (CH2)n-Y[ n which n is an integer from 0 to 4, when Y is cyclohexyl, or n is
an integer from 2 to 4, when Y is lower alkoxy having 1 to 4 carbon atom(s) or phenyl] .
In the above, the term "a saturated, straight or branched hydrocarbon radical [wherein the number of carbon atoms in the longest straight chain of this hydrocarbon radical ranges from three to seven]" preferably signifies n-propyl, l-isopropyl-2-methylpropyl, 1,1,2-trimethylpropyl, n-butyl, isobutyl, 2-ethylbutyl, 3,3-dimethylbutyl, n-pentyl, isopentyl, neopentyl, 2-propylpentyl, n-hexyl, 2-ethylhexyl, n-heptyl, allyl, 2-buten-l-yl, 3-buten-l-yl, 3-penten-l-yl, 4-penten-l-yl, 3-hexen-l-yl, 4-hexen-l-yl, 5 hexen-1-yl, and the like.
The term "a radical of the formula (CE_2)n-Y [in which n is an integer from 0 to 4, when Y is a cyclohexyl radical, or n is an integer from 2 to 4, when Y is a lower alkoxy radical having from 1 to 4 carbon atom(s) or a phenyl radical]" preferably siginifies cyclohexyl, cyclohexylmethyl,
2-cyclohexylethyl, 3-cyclohexylpropyl, 4-cyclohexylbutyl, 2-methoxyethyl, 2- ethoxyethyl, 2-propoxyethyl, 3-methoxypropyl, 3-ethoxypropyl, 4-methoxy- butyl, 4-ethoxybutyl, phenethyl, 3-phenylpropyl, 4-phenylbutyl, and the like.
Preferred pyrimidine nucleoside for the present invention are:
5'-deoxy-5-fluorouridine,
5-deoxy-5-fluorocytidine,
5'-deoxy-N -(3,5-dimethoxybenzoly)-5-fluorocytidine, 5'-deoxy-N -(3,5-dimethylbenzoly)-5-fluorocytidine,
5l-deoxy-N -[(2,4-dichlorophenyl)acetyl]-5-fluorocytidine,
5'-deoxy-N -(indol-2-ylacetyl)-5-fluorocytidine,
5'-deoxy-5-fluoro-N4-(3,4,5-trimethylbenzoly)cytidine,
5'-deoxy-5-fluoro-N4-(propoxycarbonyl)cytidine, 5'-deoxy-5-fluoro-N4-(hexyloxycarbonyl)cytidine,
5'-deoxy-5-fluoro-N4-(isopentyloxycarbonyl)cytidine,
5'-deoxy-5-fluoro-N -(neopentyloxycarbonyl)cytidine,
5'-deoxy-5-fluoro-N4- [( 1, l,2-trimethylpropoxy)carbonyl] cytidine, δ'-deoxy-N4- [(3,3-dimethylbutoxy)carbonyl] -5-fluorocytidine, 5'-deoxy-5-fluoro-N4- [( l-isopropyl-2-methylpropoxy)carbonyl] cytidine, δ'-deoxy-N4- [(2-ethylbutyl)oxycarbonyl] -5-fluorocytidine,
N4- [(cyclohexylmethoxy)carbonyl] -5'-deoxy-5-fluorocytidine,
5'-deoxy-5-fluoro-N4- [(2-phenylethoxy)carbonyl] cytidine,
2',3'-di-0-acetyl-5,-deoxy-5-fluoro-N4-(propoxycarbonyl)cytidine, 2',3'-di-O-acetyl-N4-(butoxycarbonyl)-5,-deoxy-5-fluorocytidine, 2l,3'-di-0-benzoyl-N4-(butoxycarbonyl)-5'-deoxy-5-fluorocytidine, 2,,3'-di-0-acetyl-5,-deoxy-5-fluoro-N4-(pentyloxycarbonyl)cytidine, 2',3'-di-0-acetyl-5'-deoxy-5-fluoro-N4-(isopentyloxycarbonyl)cytidine, 2,,3,-di-0-acetyl-5'-deoxy-5-fluoro-N4-(hexyloxycarbonyl)cytidine, 2,,3'-di-0-acetyl-5,-deoxy-N4-[(2-ethylbutyl)oxycarbonyl]-5-fluorocytidine, 2',3,-di-0-acetyl-N4-[(cyclohexylmethoxy)carbonyl]-5'-deoxy-5-fluorocytidine, 2',3'-di-0-acetyl-5'-deoxy-5-fluoro-N4-[(2-phenylethoxy)carbonyl]cytidine, 5'-deoxy-5-fluoro-N4-(isobutoxycarbonyl)cytidine,
5'-deoxy-5-fluoro-N4- [(2-propylpentyl)oxycarbonyl] cytidine, δ'-deoxy-N4- [(2-ethylhexyl)oxycarbonyl] -5'-fluorocytidine, δ'-deoxy-δ-fluoro-N4-(heptyloxycarbonyl)cytidine, N4-[(2-cyclohexylethoxy)carbonyl]-δ'-deoxy-δ-fluorocytidine, N4- [(3-cyclohexylpropyl)oxycarbonyl] -δ'-deoxy-δ-fluorocytidine, N4-(cyclohexyloxycarbonyl)-δ'-deoxy-δ-fluorocytidine, δ'-deoxy-δ-fluoro-N4- [(3-phenylpropyl)oxycarbonyl] cytidine, δ'-deoxy-δ-fluoro-N4- [(2-methoxyethoxy)carbonyl] cytidine, N4-(butoxycarbonyl)-δ'-deoxy-δ-fluorocytidine 5'-deoxy-5-fluoro-N4-(pentyloxycarbonyl)cytidine, 2',2'-difluorodeoxycytidine, 5-fluoro- 1-tetrahy dr ofuran- 1 -yluracil , 2'-deoxy-2'-methylidene-δ-fluorocytidine, 2'-deoxy-2'-cyano-δ-fluorocytidine and as well as hydrate or solvate thereof.
The above mentioned specific compounds are described in U.S. patent Nos. 4,071,680 and 4,966,891, European Patent Nos. 602290-A1, K. Takenuki et al. J. Med. Chem. 31, 1063 (1988), K. Yamagami et al. Cancer Research 51, 2319 (1991) and A. Matsuda et al. J. Med. Chem. 34, 2917 (1991), respectively, and those compounds can be produced according to the method described in the respective references or the analogous method thereof.
IL-12 is a heterodimeric cytokine which is produced by antigen presenting cells and serves as a pivotal regulator of T and NK cell function (cf. Stern, A.S. et al. Proc. Natl. Acad. Sci. USA. (1990) 87, 6808-6812 and Kobayashi, M. et al. J. Exp. Med. (1989) 170, 827-845). Biological activities associated with IL-12 include its ability to enhance the lytic activity of
natural killer/lymphokine activated killer cells, to induce the secretion of interferon-g (IFN-g) by both resting and activated T and NK cells, to stimulate the proliferation of activated T and NK cells, to facilitate cytotoxic T lymphocyte responses and to play a critical and unique role in promoting Th-1 type cytokine responses, thereby facilitating cell-mediated immunity (cf. Brunda, M. J. J. Leukocyte Biol. (1994) 55, 280-288 and Taniguchi, G. Blood (1994) 84, 4008-4027).
IL-12, both human type and murine type, is composed of two disulfide- bonded glycoprotein subunits approximately 35 KDa and 40 KDa in size. cDNAs encoding each subunit of IL-12 have been cloned and coexpressed in Chinese Hamster Ovary (CHO) cells to yield the secreted, bioactive, heterodimeric lymphokine (Gubler, U. et al. Proc. Natl. Acad. Sci. USA. (1991) 88, 4143-4147 and Schaenhaut, D.S. et al. J. Immunol. (1992) 148, 3433- 3440) A clone of transfected CHO cells secreting recombinant IL-12 was selected. Recombinant IL-12 was purified from the culture supernatant of CHO cells grown in a serum-free medium, by ion exchange and gel filtration chromatography.
The pharmaceutical compostion of the present invention can be administered in any form, for example, tablets, pills, suppositories, capsules, granules, powders, etc. or emulsions. The pharmaceutical composition of the present invention are especially suitable for intramuscular, subcutaneous, or intravenous administration. Pharmaceutically acceptable carriers and excipients useful in formulating the pharmaceutical composition of the present invention are those commonly used. Pharmaceutically acceptable materials can be an organic or inorganic inert carrier material suitable for enteral, percutaneous or parenteral administration such as water, gelatine, gum arabic, lactose, starch, magnesium stearate, talc, vegetable oils, polyalkylene glycols and petroleum jelly. The pharmaceutical composition provided by the present invention can be administered orally, e.g. in form of tablets, capsules, pills, powders, granules, solutions, syrups, suspensions or elixirs. The administration can also be carried out parenterally, e.g. in form of sterile solutions, suspensions or emulsions; or locally, e.g. in form of solutions, suspensions, salves, powders or aerosols. The pharmaceutical composition can be sterilized and/or can contain further adjuvants such as preserving,
stabilizing, setting, emulsifying agents, flavor-improving, salts for variation of the osmotic pressure or substances acting as buffers.
The synergistic antitumor pharmaceutical composition of the present invention comprises a single pharmaceutical composition as well as a kit of pharmaceutical compositions each containing the individual active ingredient in a desirable dosage form, thus, the present invention is also concerned with a kit for the treatment of colorectal cancer, breast cancer, stomach cancer, lung cancer, cervical cancer, bladder cancer and other malignant diseases which comprises as a first pharmaceutical composition containing an effective amount of IL-12 and a pharmaceutically acceptable carrier, and as a second pharmaceutical composition containing an effective amount of pyrimidine nucleoside derivative, as well as hydrate or solvate thereof and a pharmaceutically acceptable carrier.
Dosage ranges for the pharmaceutical composition of the present invention can easily be determined by one skilled in the art, and depend on the route of administration, the age, weight and condition of the patient and the particular disease to be treated. In the case of oral, rectal or parenteral administration for adults, an approximate range from about 0.05 mg/body/day to about 500 mg/body/day of IL-12, and about 50 mg/body/day to about 20,000 mg/body/day of pyrimidine nucleoside generally range from about 1:100 to about 1:400,000. A weight ratio from about 1:1,000 to about 1:10,000 is preferred. Rectal administration and intravenous injection are preferred routes of administration of the pharmaceutical composition according to the present invention.
The pharmaceutical compositions of the present invention are useful for the treatment of colorectal cancer, breast cancer, stomach cancer, lung cancer, cervicial cancer, bladder cancer, and other malignant diseases and the like.
The synergistic antitumor activity of the pharmaceutical composition of the present invention is evident from the tests described hereinafter.
(1) Up-regulation of the enzyme for the activation of doxifluridine and 5'- deoxy-5-fluoro-N4-(n-pentyloxycarbonyl)cytidine (capecitabine) by mIL-12.
Six weeks old female C57BL/6 mice or male BALB/c mice were inoculated with A7δδ mammary adenocarcinoma (2 x 10*^ cells) or with Meth A fibrosarcoma (5 x 10*^ cells), respectively. The mice were given s.c. mouse IL-12 (mIL-12) at 0.1 mg/mouse or vehicle (0.1 mg/ml of mouse serum albumin dissolved in phosphate-buffered saline) daily for 7 days starting at day 7 and day 8, respectively, after the tumor inoculation. One day thereafter, PyNPase activity in the tumor tissues was measured as described in Eda et al. Cancer Chemother. Pharmacol. 1993; 32:333. Mouse IFN-g (m IFN-g) levels in the tumor tissue were also measured by a commercially available ELISA system (Intertest g, Genzyme).
As Table 1 shows, mIL-12 enhanced PyNPase activity 11.9 fold in A75δ tumors and 2.4 fold in Meth A tumors. This is probably the result of the up- regulation of mIFN-g, which is an up-regulator of PyNPase.
Table 1
Up-regulation of PyNPase and mIFN-g by mIL-12
PyNPase mIFN-g
Tumor Model Administration Activity Levels
(μg δ-FU/mg/hr) (ng/g tissue)
A7δ5 mammary ca. vehicle 4.4±4.6 a) <2.0 mIL-12 52.3+28.8 * 46.8±11.7 *
Meth A fibrosarcoma vehicle 21.4±2.6 <1.1 mIL-12 51.7±9.4 * 6.7+2.1 *
* Statistically significant difference from the vehicle groups (p<0.0δ, Student t-test). a) Mean ± SD of 6 mice.
2) Selective induction of mIFN-g production by mIL-12 in the tumor tissue.
Six weeks old female C57BL/6 mice were inoculated with A7δδ mammary adenocarcinoma (2 x 105 cells). The mice were given s.c. mIL-12 at 0.1 mg/mouse or vehicle (0.1 mg/ml of mouse serum albumin dissolved in phosphate-buffered saline) daily for 7 days starting at day 6 after the tumor inoculation. One day thereafter, mIFN-g levels in the serum and tissue homogenates of tumor and other organs were measured by ELISA system as mentioned above.
As Table 2 shows, mIL-12 greatly increased mIFN-g levels in tumors as compared with those in normal organs. The tumor tissue level of mIFN-g is 3 to 7 fold higher than those of normal tissues so far examined and 50 fold higher than those in the circulation. These results suggest that mIL-12 up- regulates PyNPase selectively in the tumor tissue, through the up-regulation of mIFN-g production by IL-12 selectively in tumor tissues.
Table 2
Selective induction of mIFN-g production by mIL-12 in tumor tissues
mIFN-g levels (ng/g tissue) -l
Organs treatment of the mice vehicle mIL-12
tumor 3.2 ± 0.9 32.7 ± 9.8 *
serum 0.27 ± 0.05 0.66 ± 0.22 * small intestine <2.δ 5.3 ± 4.0 large intestine 2.8 ± 0.8 4.3 ± 0.5 * spleen 2.9 ± 0.δ 9.3 ± 1.9 * liver 2.1 ± 1.0 4.2 ± 0.6 * kidney 5.1 ± 1.1 7.5 ± 1.4 * thymus ") 6.2 10.3
* ; Significantly higher than vehicle group (p<0.05). a) ; Mean ± SD. n=4 and δ in vehicle and mIL-12 group, respectively, with an exception of thymus. °) ; Values obtained from a combined homogenate of 4 thymuses.
(3) Antitumor effects of combination of doxifluridine or capecitabine and IL-12
1) A7δδ mammary adenocarcinoma model
A7δδ (2 x 105 cells) was inoculated s.c. into female Cδ7BL/6 mice. The mice were given mIL-12 (0.03 μg/mouse, s.c), doxifluridine (1.5 mmol/kg, p.o.) and their combination, daily for 4 weeks, starting from day 9 after the tumor inoculation.
Doxifluridine as a single agent did not show activity in tumor growth inhibition because A75δ tumor has only low levels of PyNPase, whereas
mIL-12 was effective (Table 3). mIL-12 in combination with doxifluridine was much more effective than mIL-12 alone and regressed the tumor.
The combination effect was more obvious when the survival period was° observed (Table 4). Doxifluridine was not effective either in prolongation of the survival period, and IL-12 slightly prolonged the survival period. In contrast, IL-12 and doxifluridine in combination prolonged the survival period much longer than IL-12 alone and some mice were cured (3/5).
Table 3
Antitumor activity of doxifluridine, mIL-12 and their combination in A75δ mammary adenocarcinoma model.
Treatment Tumor Volume (m ^) % Tumor Growth on day 2δ a) vs. Control vs. mIL-12 alone
Control 8969 ± 4169 °) 100
doxifluridine 8380 ± 2907 93
mIL-12 2399 ± lδ67 c) 22 100
doxifluridine 216 ± 187 c) d) - 4 - 16
+ mIL-12
a) Tumor volume of 26 days after tumor inoculation was indicated, since thereafter death of mice in the control group were observed because of large tumor burden. The tumor volume on day 9 when the treatment initiated was δl3 ± 300 πm_3. b) Mean ± SD c) Significantly different from the control group, p < O.Oδ d) Significantly different from the mIL-12 group, p < O.Oδ
Table 4
Survival of mice treated with doxifluridine, mIL-12 and their combination in A7δδ mammary adenocarcinoma model
Treatment Median Survival Increase in Survivors
Days (range) Life Span % on Day lδ3
Control 28 (23 - 42) 0 o/δ doxifluridine 29 (22 - 36) 4 0/5
mIL-12 49 (43 - 55) a-* 7δ 0/5
doxifluridine
+ mIL-12 >153 (δl - >lδ3) a) ) >446 3/5
a) Significantly different from the control group, p < 0.05 b) Significantly different from the mIL-12 group, p < 0.05
2) Meth A fibrosarcoma model
Meth A fibrosarcoma (5 x 105 cells) was inoculated s.c. into male BALB/c mice. The mice were given doxifluridine (O.δ mmol/kg, p.o.), capecitabine (1.0 mmol/kg, p.o.), δ-fluorouracil (0.07δ mmol kg, p.o.), mIL-12 (0.03 μg/mouse, s.c), and their combination, daily for 3 weeks, starting from day 8 after the tumor inoculation.
Three fluoropyrimidines or mIL-12 as single agents showed moderate activity in tumor growth inhibition, at the doses employed. mIL-12 in combination with either doxifluridine or capecitabine showed more potent antitumor activity than either drug alone (p < O.Oδ). On the other hand, mIL-12 and δ-fluorouracil in combination was only slightly more effective than either drug alone (not statistically significant).
Table 5
Antitumor activity of three fluoropyrimidines (doxifluridine, capecitabine, 5-fluorouracil), mIL-12 and their combination in Meth A fibrosarcoma model.
Treatment Tumor Volume (mm3) % Tumor Growth on day 29 a) vs. Control vs. mIL-12
Control 10489 ± 2054 b) 100 doxifluridine 5170 ± 887 c) 48 - capecitabine 4941 ± 1397 c) 46 -
5-fluorouracil 6168 ± 530 c) 58 - mIL-12 4584 ± 1198 °) 42 100 mIL-12 + doxifluridine 1912 ± 1322 cde) 16 39 mIL-12 + capecitabine 2199 ± 1329 cde) 19 45 mIL-12 + 5-fluorouracil 3503 ± 1785 ∞) 32 75
a) Tumor volume of 29 days after tumor inoculation was indicated. The mean tumor volume on day 8 when the treatment initiated was 230 mm3. b) Mean ± SD c) Significantly different from the control group, p < 0.05 d) Significantly different from the mIL-12 alone group, p < 0.05 e) Significantly different from the corresponding fluoropyrimidine alone group, p < 0.05
The following examples illustrate a pharmaceutical preparation of the present invention and do not limit the scope of the present invention.
Example 1
An injectable solution containing the following ingredients was manufactured in a conventional manner: doxifluridine 1000 mg hIL-12 50 μg
NaCl 41.4 mg
NaH2P04 16.2 mg
Na2HPθ4 36.7 mg polysorbate 80 4 mg
adjust pH 7.0 with 1.2 N HC1 or 1 N NaOH adjust total volume of 20 ml with distilled water for injection
Example 2
An injectable solution each containing the following ingredients was manufactured in a conventional manner: capecitabine 100 mg hIL-12 δO μg
NaCl 718.2 mg
NaH2P04 81 mg
Na2HP04 188 mg polysorbate 80 20 mg
adjust pH 7.0 with 1.2 N HC1 or 1 N NaOH adjust total volume of 100 ml with distilled water for injection
Example 3
A kit having the following components A and B for treatment of colorectal cancer was manufactured in a conventional manner: Component A
(granule for oral administration) capecitabine 150 mg hydroxypropylmethyl cellulose 2910 4.5 mg crystalline cellulose 14.7 mg croscarmellose sodium (Ac-Di-Sol) 6.0 mg magnesium stearate 1.8 mg coating agent 3.0 mg
Total 180 mg
Component B
(sterile solution for injection) hIL-12 0.2-20 mg
NaCl 116 mg
NAH2PO4 62.2 mg Na2HP04 115.8 mg polysorbate 80 4 mg
adjust pH to 7.0 with 1.2 N HC1 or IN NaOH adjust total volume to 20 ml with water for injection
Claims
1. A synergistic antitumor pharmaceutical composition comprising an effective amount of interleukin- 12 and a pyrimidine nucleoside derivative as well as a hydrate or solvate thereof that is converted into fluorouracil or its derivative, and a pharmaceutically acceptable carrier.
2. The composition of claim 1, wherein the pyrimidine nucleoside is an uridine, cytidine or its derivative represented by the follwoing formula (I) or (II), respectively
wherein Rl is hydrogen or a radical which is easily hydrolyzable under physiological conditions; R-2 is hydrogen, cyano, fluorine, lower alkyl or lower alkylidene which may be substituted with one or two fluorine atom(s), or OR*; and R*3 is lower alkyl, hydroxymethyl, or CH2OR1, as well as hydrate or solvate thereof.
3. The composition of claim 2, wherein the easily hydrolyzable radical is R4CO-, R OCO- or R SCO-, wherein R4 is alkyl, cycloalkyl, aralkyl or aryl.
4. The composition of claim 3, wherein the alkyl, cycloalkyl, aralkyl or aryl radical represented by R4 is a saturated, straight or branched hydrocarbon radical [wherein the number of carbon atoms in the longest straight chain of this hydrocarbon radical ranges from three to seven], or a radical of the formula (CH2)n-Y[ which n is an integer from 0 to 4, when Y is cyclohexyl, or n is an integer from 2 to 4, when Y is lower alkoxy having 1 to 4 carbon atom(s) or phenyl]. δ. The composition of claim 2, wherein the pyrimidine nucleoside derivative is selected from the group consisting of:
δ'-deoxy-δ-fluorouridine, 5-deoxy-5-fluorocytidine,
5'-deoxy-N4-(3,5-dimethoxybenzoly)-δ-fluorocytidine, δ'-deoxy-N4-(3,δ-dimethylbenzoly)-δ-fluorocytidine, δ'-deoxy-N4- [(2,4-dichlorophenyl)acetyl] -δ-fluorocytidine, δ'-deoxy-N4-(indol-2-ylacetyl)-δ-fluorocytidine, δ'-deoxy-δ-fluoro-N4-(3,4,δ-trimethylbenzoly)cytidine, δ'-deoxy-5-fluoro-N4-(propoxycarbonyl)cytidine, 5'-deoxy-δ-fluoro-N4-(hexyloxycarbonyl)cytidine, δ'-deoxy-δ-fluoro-N4-(isopentyloxycarbonyl)cytidine, 5'-deoxy-5-fluoro-N4-(neopentyloxycarbonyl)cytidine, δ'-deoxy-δ-fluoro-N4- [( 1, l,2-trimethylpropoxy)carbonyl] cytidine, δ'-deoxy-N4-[(3,3-dimethylbutoxy)carbonyl]-δ-fluorocytidine, δ'-deoxy-δ-fluoro-N4- [( l-isopropyl-2-methylpropoxy)carbonyl] cytidine, δ'-deoxy-N4-[(2-ethylbutyl)oxycarbonyl]-δ-fluorocytidine, N4-[(cyclohexylmethoxy)carbonyl]-δ'-deoxy-δ-fluorocytidine, 5'-deoxy-5-fluoro-N4-[(2-ρhenylethoxy)carbonyl] cytidine,
2l,3,-di-O-acetyl-δ'-deoxy-δ-fluoro-N4-(propoxycarbonyl)cytidine, 2',3'-di-O-acetyl-N4-(butoxycarbonyl)-δ'-deoxy-δ-fluorocytidine, 2',3'-di-0-benzoyl-N4-(butoxycarbonyl)-δ'-deoxy-δ-fluorocytidine, 2',3'-di-0-acetyl-δ'-deoxy-δ-fluoro-N4-(pentyloxycarbonyl)cytidine, 2',3'-di-0-acetyl-δ'-deoxy-δ-fluoro-N4-(isopentyloxycarbonyl)cytidine, 2',3'-di-0-acetyl-δ'-deoxy-δ-fluoro-N4-(hexyloxycarbonyl)cytidine, 2',3,-di-O-acetyl-δ,-deoxy-N4-[(2-ethylbutyl)oxycarbonyl]-δ-fluorocytidine, 2',3l-di-O-acetyl-N4-[(cyclohexylmethoxy)carbonyl]-δ'-deoxy-δ-fluorocytidine, 2',3'-di-0-acetyl-δ'-deoxy-δ-fluoro-N4-[(2-phenylethoxy)carbonyl]cytidine, δ'-deoxy-δ-fluoro-N4-(isobutoxycarbonyl)cytidine, δ'-deoxy-δ-fluoro-N4- [(2-propylpentyl)oxycarbonyl] cytidine, δ'-deoxy-N4-[(2-ethylhexyl)oxycarbonyl]-5'-fluorocytidine,
5'-deoxy-δ-fluoro-N4-(heptyloxycarbonyl)cytidine,
N4- [(2-cyclohexylethoxy)carbonyl] -δ'-deoxy-δ-fluorocytidine, N4- [(3-cyclohexylpropyl)oxycarbonyl] -δ'-deoxy-δ-fluorocytidine, N -(cyclohexyloxycarbonyl)-δ'-deoxy-δ-fluorocytidine, δ'-deoxy-δ-fluoro-N4- [(3-phenylpropyl)oxycarbonyl] cytidine, δ'-deoxy-δ-fluoro-N4-[(2-methoxyethoxy)carbonyl]cytidine, N4-(butoxycarbonyl)-δ'-deoxy-δ-fluorocytidine, δ'-deoxy-δ-fluoro-N4-(pentyloxycarbonyl)cytidine, 2' ,2 '-difluorodeoxycytidine , δ-fluoro-1-tetrahydrofuran-l-yluracil, 2'-deoxy-2'-methylidene-δ-fluorocytidine, 2'-deoxy-2'-cyano-δ-fluorocytidine as well as hydrates or solvates thereof.
6. The composition any one of claims 1 to δ, wherein the weight ratio of interleukin- 12 to pyrimidine nucleoside is from about 1:100 to about 1:400,000.
7. The composition any one of claims 1 to δ, wherein the weight ratio of interleukin- 12 to pyrimidine nucleoside is from about 1:1,000 to about 1:10,000.
8. A synergistic antitumor pharmaceutical composition for the treatment of colorectal cancer, brest cancer, stomach cancer, lung cancer, cervical cancer, bladder cancer and other malignant diseases comprising an effective amount of interleukin-12 and a pyrimidine nucleoside as well as a hydrate or solvate thereof, that are converted into fluorouracil or other active metabolite by pyrimidine nucleoside phosphorylase, and a pharmaceutically acceptable carrier.
9. The composition of claim 8, wherein the pyrimidine nucleoside is an uridine, cytidine or its derivative represented by the following formula (I) or
(II), respectively as defined in claim 2 as well as a hydrate or solvate thereof.
10. The composition of claim 8, wherein the easily hydrolyzable radical is R4CO-, R4OCO- or R4SCO-, wherein R4 is alkyl, cycloalkyl, aralkyl or aryl.
11. The composition of claim 10, wherein the alkyl, cycloalkyl, aralkyl or aryl radical represented by R4 is a saturated, straight or branched hydrocarbon radical [wherein the number of carbon atoms in the longest straight chain of this hydrocarbon radical ranges from three to seven], or a radical of the formula (CH2)n-Y[ which n is an integer from 0 to 4, when Y is cyclohexyl, or n is an integer from 2 to 4, when Y is lower alkoxy having 1 to 4 carbon atom(s) or phenyl] .
12. The composition of claim 9, wherein the pyrimidine nucleoside derivative is selected from the compounds defined in claim 5.
13. The composition any one of claims 8 to 12, wherein the weight ratio of interleukin- 12 to pyrimidine nucleoside is from about 1:100 to about 1:400,000.
14. The composition any one of claims 8 to 12, wherein the weight ratio of interleukin- 12 to pyrimidine nucleoside is from about 1:1,000 to about 1:10,000.
lδ. A method of treating colorectal cancer, brest cancer, stomach cancer, lung cancer, cervical cancer, bladder cancer and other malignant diseases which comprises administering a synergistic antitumor pharmaceutical composition comprising an effective amount of interleukin- 12 and pyrimidine nucleoside or as well as hydrate or solvate thereof, that are converted into fluorouracil or other active metabolite by pyrimidine nucleoside phosphorylase, and a pharmaceutically acceptable carrier.
16. The method of claim lδ, wherein the pyrimidine nucleoside is an uridine, cytidine or its derivative represented by the following formula (I) or (II), respectively, as defined in claim 2, as well as a hydrate or solvate thereof.
17. The method of claim 16, wherein the easily hydrolyzable radical is R4CO-, R4OCO- or R4SCO-, wherein R4 is alkyl, cycloalkyl, aralkyl or aryl.
18. The method of claim 17, wherein the alkyl, cycloalkyl, aralkyl or aryl radical represented by R4 is a saturated, straight or branched hydrocarbon radical [wherein the number of carbon atoms in the longest straight chain of this hydrocarbon radical ranges from three to seven], or a radical of the formula (CH2)n-Y -in which n is an integer from 0 to 4, when Y is cyclohexyl, or n is an integer from 2 to 4, when Y is lower alkoxy having 1 to 4 carbon atom(s) or phenyl].
19. The composition of claim lδ, wherein the pyrimidine nucleoside derivative is the one selected from the compounds defined in claim δ, as well as a hydrate or solvate thereof.
20. The method any one of claims lδ to 19, wherein the weight ratio of interleukin- 12 to pyrimidien nucleoside is from about 1:100 to about 1:400,000.
21. The method any one of claims lδ to 19, wherein the weight ratio of interleukin-12 to pyrimidine nucleoside is from about 1:1,000 to 1:10,000.
22. A kit for the treatment of colorectal cancer, breast cancer, stomach cancer, lung cancer, cervical cancer, bladder cancer and other malignant diseases which comprises as a first pharmaceutical composition containing an effective amount of interleukin- 12 and a pharmaceutically acceptable carrier, and as a second pharmaceutical composition containing an effective amount of a pyrimidine nucleoside derivative, as well as a hydrate or solvate thereof and a pharmaceutically acceptable carrier.
23. The use of interleukin- 12 and a pharmaceutically acceptable carrier, and as a second pharmaceutical composition containing an effective amount of a pyrimidine nucleoside derivative, as well as a hydrate or solvate thereof for the manufacture of pharmaceutical compositions as claimed in claims 8 or 22.
***
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| EP96917379A EP0828508A2 (en) | 1995-05-26 | 1996-05-15 | Compositions of interleukin and pyrimidine nucleosides |
Applications Claiming Priority (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| EP95108063 | 1995-05-26 | ||
| EP95108063 | 1995-05-26 | ||
| PCT/EP1996/002088 WO1996037214A2 (en) | 1995-05-26 | 1996-05-15 | Compositions of interleukin and pyrimidine nucleosides |
| EP96917379A EP0828508A2 (en) | 1995-05-26 | 1996-05-15 | Compositions of interleukin and pyrimidine nucleosides |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| EP0828508A2 true EP0828508A2 (en) | 1998-03-18 |
Family
ID=8219298
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| Application Number | Title | Priority Date | Filing Date |
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| EP96917379A Withdrawn EP0828508A2 (en) | 1995-05-26 | 1996-05-15 | Compositions of interleukin and pyrimidine nucleosides |
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| Country | Link |
|---|---|
| EP (1) | EP0828508A2 (en) |
| JP (1) | JPH10506641A (en) |
| CN (1) | CN1184430A (en) |
| AR (1) | AR005417A1 (en) |
| AU (1) | AU5998296A (en) |
| BR (1) | BR9608779A (en) |
| CA (1) | CA2220837A1 (en) |
| TR (1) | TR199701435T1 (en) |
| WO (1) | WO1996037214A2 (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0882734B1 (en) * | 1997-06-02 | 2009-08-26 | F. Hoffmann-La Roche Ag | 5'-Deoxy-cytidine derivatives |
| NZ330360A (en) * | 1997-06-02 | 1999-03-29 | Hoffmann La Roche | 5'-deoxy-cytidine derivatives, their manufacture and use as antitumoral agents |
-
1996
- 1996-05-15 CN CN96194016A patent/CN1184430A/en active Pending
- 1996-05-15 CA CA002220837A patent/CA2220837A1/en not_active Abandoned
- 1996-05-15 EP EP96917379A patent/EP0828508A2/en not_active Withdrawn
- 1996-05-15 JP JP8535342A patent/JPH10506641A/en active Pending
- 1996-05-15 TR TR97/01435T patent/TR199701435T1/en unknown
- 1996-05-15 BR BR9608779A patent/BR9608779A/en not_active Application Discontinuation
- 1996-05-15 WO PCT/EP1996/002088 patent/WO1996037214A2/en not_active Application Discontinuation
- 1996-05-15 AU AU59982/96A patent/AU5998296A/en not_active Abandoned
- 1996-05-22 AR ARP960102652A patent/AR005417A1/en unknown
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| Title |
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| See references of WO9637214A2 * |
Also Published As
| Publication number | Publication date |
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| WO1996037214A3 (en) | 1997-01-09 |
| AR005417A1 (en) | 1999-06-23 |
| CA2220837A1 (en) | 1996-11-28 |
| AU5998296A (en) | 1996-12-11 |
| TR199701435T1 (en) | 1998-03-21 |
| BR9608779A (en) | 1999-07-06 |
| WO1996037214A2 (en) | 1996-11-28 |
| MX9708915A (en) | 1998-03-31 |
| CN1184430A (en) | 1998-06-10 |
| JPH10506641A (en) | 1998-06-30 |
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