CA2237695A1 - Antineoplastic methods and compositions employing optically pure (-)-fotemustine - Google Patents
Antineoplastic methods and compositions employing optically pure (-)-fotemustine Download PDFInfo
- Publication number
- CA2237695A1 CA2237695A1 CA002237695A CA2237695A CA2237695A1 CA 2237695 A1 CA2237695 A1 CA 2237695A1 CA 002237695 A CA002237695 A CA 002237695A CA 2237695 A CA2237695 A CA 2237695A CA 2237695 A1 CA2237695 A1 CA 2237695A1
- Authority
- CA
- Canada
- Prior art keywords
- fotemustine
- otemustine
- optically pure
- cells
- malignancy
- 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.)
- Abandoned
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/66—Phosphorus compounds
- A61K31/662—Phosphorus acids or esters thereof having P—C bonds, e.g. foscarnet, trichlorfon
-
- 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
- A61P35/00—Antineoplastic agents
- A61P35/04—Antineoplastic agents specific for metastasis
Landscapes
- Health & Medical Sciences (AREA)
- Public Health (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Chemical & Material Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Organic Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Oncology (AREA)
- Epidemiology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
Methods and compositions are disclosed utilizing optically pure (-)-fotemustine for the treatment of various malignancies, while substantially reducing the concomitant liability of adverse effects associated with the racemic mixture of fotemustine.
Description
CA 0223769~ 1998-0~-14 WO97/1881g PCT~S96/18695 ANTINEOP~ASTIC METHODS AND COMPOSITIONS EMPLOYING
OPTICA~LY PURE (-)-FOTEhu~ 'l'LN~
FIE~ OF T~E INVENTIQN
This invention relates to pharmaceutical compositions cont~'n~ng (~ otemustine. In another aspect this invention relates to treatment o~
malignancies, including lymphomas, melanomas, gliomas and other brain tumors, lung cancer, liver metastases and hematological and gastrointestinal malignancies.
BACKGROUND OF TH~ INVENTION
In the standard nomenclature o~ Chemical Abstracts, ~otemustine is [l-[[[~2-chloroethyl) nitosoamino~carbonyl~amino]ethylphosphonic acid diethyl ester. Fotemustine is also known as 1-[N-(2-chloroethyl)-N-nitrosoureido~ethylphosphonic acid diethyl ester and as diethyl-1-[3-(2-chloroethyl)-3-nitrosoureido]ethylphosphonate. Early re~erences re~er to ~otemustine as S10036. The preparation o~
racemic ~otemustine is described in U.S. Patent 4,567,169. The patent mentions that "the invention also relates to the optical isomers o~ the compounds...", but no such isomers are disclosed.
Racemic rotemus~ine has been used 'clinically to treat malignan~ melanoma, ~rimary and metastatic brain tumors, colorectal cancer, so~t tissue and bore sarcomas and hematological malisnancies. [See .~hayat et al. J. Nat'l. Cancer Inst. 80, 1407-1408 (1988);
Khayat et al. Cancer Res. 47, 6782-6785 (1987);
Jacquillat et al. Proc. Am. Assoc. C~cer Res. 30, CA 0223769~ l998-0~-l4 WO97/18819 PCT~S96/18695 A1088 (1989); Rougier et al. Eur. J. Cancer 2~A~2), 288-9 (1993); Kerbrat et al. Eur J. Cancer 2~Arl), 143-144 ~1993)]. These clinical studies indicate that racemic fotemustine is clinically use~ul in treating these cancers, ~ut its use is accompanied by delayed, cumulative toxicity in bone marrow. The dose-limiting toxicity is usually myelosuppression leading to neutropenia, leucopenia and thrombocytopenia. Renal, pulmonary and hepatic toxicity are observed with a lesser frequency.
Racemic ~otemustine is also believed to be mutagenic ~See Ashby et al. Mutation Res. 286, lO1-109 (~993)].
Nausea is a problem in some cases.
It would be particularly desirable to find a compound with the advantages of the racemic mixture of fotemustine but without the side effects enumerated above.
SUMMARY OF THF lN V~N 1 ION
It has now been discovered that the optically pure (-) isomer of ~otemustine is an ef~ective agent ~or treating malignancies. The optically pure (-) isomer of fotemustine provides this effective treatment while substantially reducing one or more ad~erse ef~ects associated with administration of racemic ~otemustine including, but not limited to, neu~ropenia, leucopenia ard thrombocytopenia; renal, ~ulmonary and hepatic toxicity; mutagenicity; and nausea.
In one aspect the invention relates to a method o~ treating a malignancy which comprises .
.
WO97/18819 PCT~S96/1869 administering to a ~mm~ 1 su~ering ~rom the malignancy, a therapeutically e~ective amount o~
c (~ otemustine, substantially ~ree o~ its (+) stereoisomer.
In another aspect, the invention relates to a pharmaceutical composition which comprises (-)-~otemustine, substantially free of its (+) stereoisomer, and a pharmaceutically acceptable carrier.
DET~ILED DESCRIPTION OF THE INVENTION
The active compound used in the compositions and methods o~ the invention is the levorotatary or (-) optical isomer of ~otemustine. The absolute stereochemistry o~ the (-) isomer is presently unknown. Formula I indicates a single enanticmer o~
undetermined absolute stereochemistry, here presented arbitrarily as S~:
E t 3 3~ ~ ~l o E; C~ ~ N~
The sraphic representation o~ enantiomerically pure ~otemustine is taken from Maehr J. Chem. E-. 62, ll~-120 (1985). Thus representations involvins wedge outlines and dotted or broken lines denote CA 0223769~ 1998-0~-14 WO97/18819 PCT~S96/18695 enantiomerically pure compounds of indeterminate absolute configuration.
The present invention encompasses a method of treating a malignancy which comprises administering a 5 therapeutically effective amount of ~-)-fotemustine, substantially free of its (+) stereoisomer.
The term "substantially ~ree of its (+) stereoisomer" as used herein means that the compositions contain at least 9O~ by weight of (-)-~otemustine and lO~ by weight or less of (+) ~otemustine. In a more preferred embodiment the term llsubstantially free of the (+) isomer" means that the composition contains at least 99~ by weight of (-)-fotemustine, and l~ or less o~ (+) fotemustine. In the most preferred embodiment, the term "substantially free o its (+) stereoisomer" as used herein means that the composition contains greater than 99~ by weight o~ fotemustine. These percentages are based upon the total amount of fotemustine in the composition. The terms "substantially optically pure (-) isomer of fotemustine" or "substantially optically pure (-)-fotemustine" and "optically pure (-) isomer of fotemustinen and ~loptically pure (-)-fotemustine" are also encompassed by the abov~-described amounts.
The term "treatlng a malignancyl~ as used herein means treating, alleviating or palliating such condition, suppressing the growth o~ cancerous tissue and thus providing increased survival time.
CA 0223769~ 1998-0~-14 WO97/18819 PCT~S96/18695 The term "therapeutically effective amount"
refers to that amount of ~-)-fotemustine which is sufficient to treat a malignancy. Malignancies against which (-) fotemustine is active include lymphomas, melanomas, gliomas and other brain tumors, lung cancer, liver metastases and hematological and gastrointestinal malignancies.
The method of the invention reduces the concomitant liability of adverse effects associated with the administration of racemic fotemustine. The term "adverse effects" includes, but is not limited to, neutropenia, leucopenia and thrombocytopenia;
renal, pulmonary and hepatic toxicity; mutagenicity;
and nausea.
In the method of the invention a therapeutically e~fective amount may be ~m;n~ stered in a single dose, or a ~raction of a therapeutically effective amount may be ~mi n~ stered in each of several doses.
hikewise a pharmaceutical composition of the invention may contain a therapeutically e~fective amount of (-)-fotemustine or it may be necessary to ~mi n, ster several individual compositions in order to achieve therapeutic effectiveness.
The amount of (-)-fotemustine that constitutes a therapeutically effective amount varies with the severity and nature of the condition to be treated and the route of administration. Dose size and dose ~requency may also vary according to the age, body weight and response of the individual patient. In general, the dose range for a single administration of (-)-~otemustine for the conditions described CA 0223769~ 1998-0~-14 WO97/18819 PCT~S96/18695 herein is from about 20 mg/kg to about 150 mg/kg by intravenous or intraarterial in~usion. Preferably a single dose range should be about 20 mg/m2 to about 80 mg/m2. Generally, dosing is carried out periodically (e.g., weekly) ~or a period o~ several weeks (e.g., about two to about to about eight weeks).
In managing the patient, therapy should be initiated at a lower dose, perhaps at about 20 mg/m2 to about 40 mg/m2, and increased up to about 40 mg/m~
or higher depending on the patient's global response.
It is ~urther reco~m~n~ed that patients over 65 years of age and those with impaired renal or hepatic ~nction initially receive low doses and that they be titrated based on individual response(s) and blood level (8) . It may be necessary to use dosages outside these ranges in some cases, as will be apparent to those skilled in the art. Further, the clinician or treating physician will know how and when to interrupt, adjust, or terminate therapy in conjunction with individual patient response. The term "an amount su~ficient to suppress cancer but insu~icient to cause said adverse e~ects" is encompassed by the above-described dosage amounts and dose ~requency schedule.
2s Any sui~able route of administration may be employed ~or providing the patient with an efIective dosage of (-)-fotemustine, but parenteral (particularly intravenous and intraarterial) forms of adminis~ration are preferred Dosage forms ir.clude dispersions, suspensions, solutions, and the like.
CA 0223769j l998-oj-l4 W~97/18819 PCT~S96/~8695 The pharmaceutical compositions of the present invention comprise (-)-fotemustine as the active ingredient, and may also contain a pharmaceutically acceptable carrier, and optionally, other adjuvants, 5 excipients, or therapeutic ingredients. Compositions suitable for parenteral administration may be presented as an a~ueous solution, optionally containing conventional adjuvants, and excipients.
Such compositions may be prepared by methods well 10 ~nown to those s~illed in the art of pharmacy. All methods include the step o~ bringing into association the active ingredient with the carrier which constitutes one or more necessary ingredients.
The chemical synthesis of the racemic mixture of 15 fotemustine can be performed by the method described in U.S. Patent 4,567,169 cited above. The (-) isomer o~ fotemustine may then ~e resolved by chromatography on a chiral medium. Alternatively, (-)-fotemustine may be obtained by resolution of the enantiomers of 20 the amine precursor to fotemustine using fractional crystallization or chromatography of diastereomeric esters of chiral acids. Other standard methods of resolution known to those skilled in the art can also be used. (See for example, E.L. Eliel, 25 Stereochemistry of Carbon Compounds, McGraw Hill ~19~2) and [Wilen and Lochmuller, "Tables of Resolving Agents", Journal of ChrQmatoqra~h~ 113 t 28~-302 (1975)].
,, Relative activity, potency and specificity of r 30 optically pure fotemustine and racemic fotemustine as an antineoplastic agent can be determined by pharmacological studies in vi tro and in vivo .
WO97/18819 PCT~S96/18695 The compounds are tested for their capacity to increase the life-span of mice bearing tumorous cells inoculated by intraperitoneal or intramuscular route in accordance with the protocols established by the National Cancer Institute (U.S.A.) and published by R.I. Geran et al. in ~ancer ChemotheraPY Re~orts, part III, Vol. 3(2), 1-87 (1972).
Hematopoietic toxicity_is assessed, in animals treated with one or several administrations of the compound according to the invention, by the count of the peripheral blood cells and of the bone marrow, and the stem cells contained in the bone marrow [Till and McCulloch, Radiation Res. 14, 213 (1961)]. The lowest cellular concentrations are noted 3 days after the start of treatment. The extent of this decrease is measured a~ter treatment with one dose of N-N~-bis(2-chloroethyl)-N-nitrosourea ("BCNU") used as a reference tTang and Eisenbrand, Arch. Pharm. 314, 9lO
(1981)~ and is compared to the cell count after racemic fotemustine and (+) fotemustine.
Hepatic toxicity is assessed according to Wroblewski's method by measuring the pyruvic glutamic trans~ se activity in serum from Long Evans rats treated with an intraperitoneal injection o~ doses of racemic, ~+) and (-) ~otemustine.
Mutagenicity is assessed Dy the method of Tapiero et al. ~Anticancer Research 9, 1617-1622 (1989)] as follows:
Human lung carcinomas A427 and A549 and human colon carcinomas BE and HT29 are obtained and CA 0223769~ 1998-0~-14 WO97/18819 PCT~S96/1869S
maintained as described by Kohn et al. rCancer Chemother. Pharmacol 19, 291-295 (1987) and Cancer Res. 48, 3622-3625 (1988)]. Murine leukemia cell line P388 is obtained from NCI. Cells are grown and maintained in RPMI 1640 medium supplemented with lO~
~etal bovine serum, lO-sM 2-mercaptoethanol and standard antibiotics. All cultures are grown at 38~
in a humidified atmosphere o~ 5~ CO2. Cell growth inhibition studies are carried out on exponential cultures in the continuous presence of drug.
Resultant cell numbers are estimated on a Coulter Counter model ZBI, Coulter Electronics, Hialeah, Fl.
Racemic, (+~ and (-) fotemustine are individually dissolved in ethanol immediately be~ore use. Convenient standard stock solutions are 35 mM.
For radioisotope labelling and X-irradiation 2.5X lOs P388 cell/mL are grown in medium cont~in;ng either O.05~Ci/mL of (14C)-thymidine or 0.5~Ci/mL of (3H)-thymidine. After labelling for 20 hours, medium is removed and cells washed with cold PBS. Standard chase treatments using lO~M non-radioactive th~midine ~or 4 hours are employed. Tritiated cells are expo~ed, on ice, to 300 rads of X-irradiation as an internal standard. The alkaline elution procedure is described by Kohn e~ al. ~Measurements of strand breaks and cross l~nkc by alkalire elut~on." In:
Frieberg E. and Hanawalt P. (eds), DNA repair: A
laboratory m~n1l~l of research procedures, New York, ' Dekker, l: 1981, pp. 379-401.] To determine the relationship between preincubation of drug in complete growth medium and loss of cytotoxic activity, drugs are resuspended in medium containing lO~ fetal calf serum and incubated at 37O for l to WO97/18819 PCT~S96/18695 500 minutes. P388 cells are resuspended in the medium diluted to reach the appropriate drug concentrations. After 3 days incubation at 5~ CO2, cells are counted.
Induction of single strand breaks in P388 cells exposed to racemic fotemustine, ~+) fotemustine and ~ otemus~ine i5 compared. Cells (5XlO~) labelled 20 hours by (l4C)-thymidine are treated ~or 2 hours with various concentrations of ~otemustine and mixed with a similar number of (3H)-thymidine labelled reference cells. These cells are collected on 2.0 ~m pore size polycarbonate ~ilters and lysed with 5 mL
o~ 2~ SDS/0.025 M Na4EDTA ~pH9.7) in the presence o~
proteinase K. Elution is carried out in the dark at pH 12.l as described by Kohn. DNA single strand break ~requencies are calculated on the basis of ~irst order elution kinetics.
Induction of DNA-Protein cross-links is examined in P388 cells treated ~or various times in the presence of racemic fotemustine, (+) ~otemustine or ~-) fotemustine. (l4C)-Thymidine-labelled drug treated cells are irradiated with 600 rads o~ X-rays and (3H)-thymidine labelled reference cells are irradiated with 300 rads of X-rays. These cells are collected on 2.0~m pore-size polycarbonate ~ilters and lysed at pH 9.7 in the absence of proteinase K. Elution is the same as above.
Removal of DNA single strand breaks in ~ 388 cells at various times after exposure to concentrations of racemic fotemustine and its enantiomers causing equivalent DNA damage can also be _ CA 0223769~ 1998-0~-14 WO97/18819 PCT~S96/18695 m; n~d. The (l~C)-thymidine labelled cells initially exposed to drug treatment ~or 2 hours are washed and resuspended in drug- f ree medium. A~ter di~ferent times of incubation at 37~, these cel~s are mixed with an equal number of ~3H)-thymidine labelled reference cells, collected on the polycarbonate ~ilters and analyzed by alkaline elution.
The ~oregoing tests provide an estimate o~
relative activity, potency and selectivity.
Emesis in Ferrets. Experiments are performed on adult male ~errets. The ~im~l S are ~irst adapted to wearing a nylon jacket connected to a stainless-steel cable, which in turn is attached to a brass swivel at the cage top. After habituation to the tether-harness, each ~n;m~l receives a surgically implanted catheter in its right jugular vein. The catheter is ~lushed daily with heparinized sodium chloride. The drug studies are conducted l week a~ter the surgical procedure. Tethered animals are individually housed.
Eight to eleven ~nim~l s are used to evaluate each dose of each test compound. Individual animals are weighed weekly and randomly given, at greater than 48 hour intervals, a single i.v. or p.o. dose of racemic ~otemustine, (+)-~otemustine or (-)-~otemustine. At least three dose levels o~ each test compound are evaluated.
Individual animals are observed ~or 30 minutes ~ollowing administration o~ the test substance. The ~requency of, and latency to all expulsions, retches WO97/18819 PCT~S96/18C95 and de~ecations are recorded. Data obtained ~rom dose-response curves is tested ~or statistical significance by chi-square analysis. EDso values are determined ~or each compound. The test provides an estimate o~ relative liability to nausea and vomiting.
OPTICA~LY PURE (-)-FOTEhu~ 'l'LN~
FIE~ OF T~E INVENTIQN
This invention relates to pharmaceutical compositions cont~'n~ng (~ otemustine. In another aspect this invention relates to treatment o~
malignancies, including lymphomas, melanomas, gliomas and other brain tumors, lung cancer, liver metastases and hematological and gastrointestinal malignancies.
BACKGROUND OF TH~ INVENTION
In the standard nomenclature o~ Chemical Abstracts, ~otemustine is [l-[[[~2-chloroethyl) nitosoamino~carbonyl~amino]ethylphosphonic acid diethyl ester. Fotemustine is also known as 1-[N-(2-chloroethyl)-N-nitrosoureido~ethylphosphonic acid diethyl ester and as diethyl-1-[3-(2-chloroethyl)-3-nitrosoureido]ethylphosphonate. Early re~erences re~er to ~otemustine as S10036. The preparation o~
racemic ~otemustine is described in U.S. Patent 4,567,169. The patent mentions that "the invention also relates to the optical isomers o~ the compounds...", but no such isomers are disclosed.
Racemic rotemus~ine has been used 'clinically to treat malignan~ melanoma, ~rimary and metastatic brain tumors, colorectal cancer, so~t tissue and bore sarcomas and hematological malisnancies. [See .~hayat et al. J. Nat'l. Cancer Inst. 80, 1407-1408 (1988);
Khayat et al. Cancer Res. 47, 6782-6785 (1987);
Jacquillat et al. Proc. Am. Assoc. C~cer Res. 30, CA 0223769~ l998-0~-l4 WO97/18819 PCT~S96/18695 A1088 (1989); Rougier et al. Eur. J. Cancer 2~A~2), 288-9 (1993); Kerbrat et al. Eur J. Cancer 2~Arl), 143-144 ~1993)]. These clinical studies indicate that racemic fotemustine is clinically use~ul in treating these cancers, ~ut its use is accompanied by delayed, cumulative toxicity in bone marrow. The dose-limiting toxicity is usually myelosuppression leading to neutropenia, leucopenia and thrombocytopenia. Renal, pulmonary and hepatic toxicity are observed with a lesser frequency.
Racemic ~otemustine is also believed to be mutagenic ~See Ashby et al. Mutation Res. 286, lO1-109 (~993)].
Nausea is a problem in some cases.
It would be particularly desirable to find a compound with the advantages of the racemic mixture of fotemustine but without the side effects enumerated above.
SUMMARY OF THF lN V~N 1 ION
It has now been discovered that the optically pure (-) isomer of ~otemustine is an ef~ective agent ~or treating malignancies. The optically pure (-) isomer of fotemustine provides this effective treatment while substantially reducing one or more ad~erse ef~ects associated with administration of racemic ~otemustine including, but not limited to, neu~ropenia, leucopenia ard thrombocytopenia; renal, ~ulmonary and hepatic toxicity; mutagenicity; and nausea.
In one aspect the invention relates to a method o~ treating a malignancy which comprises .
.
WO97/18819 PCT~S96/1869 administering to a ~mm~ 1 su~ering ~rom the malignancy, a therapeutically e~ective amount o~
c (~ otemustine, substantially ~ree o~ its (+) stereoisomer.
In another aspect, the invention relates to a pharmaceutical composition which comprises (-)-~otemustine, substantially free of its (+) stereoisomer, and a pharmaceutically acceptable carrier.
DET~ILED DESCRIPTION OF THE INVENTION
The active compound used in the compositions and methods o~ the invention is the levorotatary or (-) optical isomer of ~otemustine. The absolute stereochemistry o~ the (-) isomer is presently unknown. Formula I indicates a single enanticmer o~
undetermined absolute stereochemistry, here presented arbitrarily as S~:
E t 3 3~ ~ ~l o E; C~ ~ N~
The sraphic representation o~ enantiomerically pure ~otemustine is taken from Maehr J. Chem. E-. 62, ll~-120 (1985). Thus representations involvins wedge outlines and dotted or broken lines denote CA 0223769~ 1998-0~-14 WO97/18819 PCT~S96/18695 enantiomerically pure compounds of indeterminate absolute configuration.
The present invention encompasses a method of treating a malignancy which comprises administering a 5 therapeutically effective amount of ~-)-fotemustine, substantially free of its (+) stereoisomer.
The term "substantially ~ree of its (+) stereoisomer" as used herein means that the compositions contain at least 9O~ by weight of (-)-~otemustine and lO~ by weight or less of (+) ~otemustine. In a more preferred embodiment the term llsubstantially free of the (+) isomer" means that the composition contains at least 99~ by weight of (-)-fotemustine, and l~ or less o~ (+) fotemustine. In the most preferred embodiment, the term "substantially free o its (+) stereoisomer" as used herein means that the composition contains greater than 99~ by weight o~ fotemustine. These percentages are based upon the total amount of fotemustine in the composition. The terms "substantially optically pure (-) isomer of fotemustine" or "substantially optically pure (-)-fotemustine" and "optically pure (-) isomer of fotemustinen and ~loptically pure (-)-fotemustine" are also encompassed by the abov~-described amounts.
The term "treatlng a malignancyl~ as used herein means treating, alleviating or palliating such condition, suppressing the growth o~ cancerous tissue and thus providing increased survival time.
CA 0223769~ 1998-0~-14 WO97/18819 PCT~S96/18695 The term "therapeutically effective amount"
refers to that amount of ~-)-fotemustine which is sufficient to treat a malignancy. Malignancies against which (-) fotemustine is active include lymphomas, melanomas, gliomas and other brain tumors, lung cancer, liver metastases and hematological and gastrointestinal malignancies.
The method of the invention reduces the concomitant liability of adverse effects associated with the administration of racemic fotemustine. The term "adverse effects" includes, but is not limited to, neutropenia, leucopenia and thrombocytopenia;
renal, pulmonary and hepatic toxicity; mutagenicity;
and nausea.
In the method of the invention a therapeutically e~fective amount may be ~m;n~ stered in a single dose, or a ~raction of a therapeutically effective amount may be ~mi n~ stered in each of several doses.
hikewise a pharmaceutical composition of the invention may contain a therapeutically e~fective amount of (-)-fotemustine or it may be necessary to ~mi n, ster several individual compositions in order to achieve therapeutic effectiveness.
The amount of (-)-fotemustine that constitutes a therapeutically effective amount varies with the severity and nature of the condition to be treated and the route of administration. Dose size and dose ~requency may also vary according to the age, body weight and response of the individual patient. In general, the dose range for a single administration of (-)-~otemustine for the conditions described CA 0223769~ 1998-0~-14 WO97/18819 PCT~S96/18695 herein is from about 20 mg/kg to about 150 mg/kg by intravenous or intraarterial in~usion. Preferably a single dose range should be about 20 mg/m2 to about 80 mg/m2. Generally, dosing is carried out periodically (e.g., weekly) ~or a period o~ several weeks (e.g., about two to about to about eight weeks).
In managing the patient, therapy should be initiated at a lower dose, perhaps at about 20 mg/m2 to about 40 mg/m2, and increased up to about 40 mg/m~
or higher depending on the patient's global response.
It is ~urther reco~m~n~ed that patients over 65 years of age and those with impaired renal or hepatic ~nction initially receive low doses and that they be titrated based on individual response(s) and blood level (8) . It may be necessary to use dosages outside these ranges in some cases, as will be apparent to those skilled in the art. Further, the clinician or treating physician will know how and when to interrupt, adjust, or terminate therapy in conjunction with individual patient response. The term "an amount su~ficient to suppress cancer but insu~icient to cause said adverse e~ects" is encompassed by the above-described dosage amounts and dose ~requency schedule.
2s Any sui~able route of administration may be employed ~or providing the patient with an efIective dosage of (-)-fotemustine, but parenteral (particularly intravenous and intraarterial) forms of adminis~ration are preferred Dosage forms ir.clude dispersions, suspensions, solutions, and the like.
CA 0223769j l998-oj-l4 W~97/18819 PCT~S96/~8695 The pharmaceutical compositions of the present invention comprise (-)-fotemustine as the active ingredient, and may also contain a pharmaceutically acceptable carrier, and optionally, other adjuvants, 5 excipients, or therapeutic ingredients. Compositions suitable for parenteral administration may be presented as an a~ueous solution, optionally containing conventional adjuvants, and excipients.
Such compositions may be prepared by methods well 10 ~nown to those s~illed in the art of pharmacy. All methods include the step o~ bringing into association the active ingredient with the carrier which constitutes one or more necessary ingredients.
The chemical synthesis of the racemic mixture of 15 fotemustine can be performed by the method described in U.S. Patent 4,567,169 cited above. The (-) isomer o~ fotemustine may then ~e resolved by chromatography on a chiral medium. Alternatively, (-)-fotemustine may be obtained by resolution of the enantiomers of 20 the amine precursor to fotemustine using fractional crystallization or chromatography of diastereomeric esters of chiral acids. Other standard methods of resolution known to those skilled in the art can also be used. (See for example, E.L. Eliel, 25 Stereochemistry of Carbon Compounds, McGraw Hill ~19~2) and [Wilen and Lochmuller, "Tables of Resolving Agents", Journal of ChrQmatoqra~h~ 113 t 28~-302 (1975)].
,, Relative activity, potency and specificity of r 30 optically pure fotemustine and racemic fotemustine as an antineoplastic agent can be determined by pharmacological studies in vi tro and in vivo .
WO97/18819 PCT~S96/18695 The compounds are tested for their capacity to increase the life-span of mice bearing tumorous cells inoculated by intraperitoneal or intramuscular route in accordance with the protocols established by the National Cancer Institute (U.S.A.) and published by R.I. Geran et al. in ~ancer ChemotheraPY Re~orts, part III, Vol. 3(2), 1-87 (1972).
Hematopoietic toxicity_is assessed, in animals treated with one or several administrations of the compound according to the invention, by the count of the peripheral blood cells and of the bone marrow, and the stem cells contained in the bone marrow [Till and McCulloch, Radiation Res. 14, 213 (1961)]. The lowest cellular concentrations are noted 3 days after the start of treatment. The extent of this decrease is measured a~ter treatment with one dose of N-N~-bis(2-chloroethyl)-N-nitrosourea ("BCNU") used as a reference tTang and Eisenbrand, Arch. Pharm. 314, 9lO
(1981)~ and is compared to the cell count after racemic fotemustine and (+) fotemustine.
Hepatic toxicity is assessed according to Wroblewski's method by measuring the pyruvic glutamic trans~ se activity in serum from Long Evans rats treated with an intraperitoneal injection o~ doses of racemic, ~+) and (-) ~otemustine.
Mutagenicity is assessed Dy the method of Tapiero et al. ~Anticancer Research 9, 1617-1622 (1989)] as follows:
Human lung carcinomas A427 and A549 and human colon carcinomas BE and HT29 are obtained and CA 0223769~ 1998-0~-14 WO97/18819 PCT~S96/1869S
maintained as described by Kohn et al. rCancer Chemother. Pharmacol 19, 291-295 (1987) and Cancer Res. 48, 3622-3625 (1988)]. Murine leukemia cell line P388 is obtained from NCI. Cells are grown and maintained in RPMI 1640 medium supplemented with lO~
~etal bovine serum, lO-sM 2-mercaptoethanol and standard antibiotics. All cultures are grown at 38~
in a humidified atmosphere o~ 5~ CO2. Cell growth inhibition studies are carried out on exponential cultures in the continuous presence of drug.
Resultant cell numbers are estimated on a Coulter Counter model ZBI, Coulter Electronics, Hialeah, Fl.
Racemic, (+~ and (-) fotemustine are individually dissolved in ethanol immediately be~ore use. Convenient standard stock solutions are 35 mM.
For radioisotope labelling and X-irradiation 2.5X lOs P388 cell/mL are grown in medium cont~in;ng either O.05~Ci/mL of (14C)-thymidine or 0.5~Ci/mL of (3H)-thymidine. After labelling for 20 hours, medium is removed and cells washed with cold PBS. Standard chase treatments using lO~M non-radioactive th~midine ~or 4 hours are employed. Tritiated cells are expo~ed, on ice, to 300 rads of X-irradiation as an internal standard. The alkaline elution procedure is described by Kohn e~ al. ~Measurements of strand breaks and cross l~nkc by alkalire elut~on." In:
Frieberg E. and Hanawalt P. (eds), DNA repair: A
laboratory m~n1l~l of research procedures, New York, ' Dekker, l: 1981, pp. 379-401.] To determine the relationship between preincubation of drug in complete growth medium and loss of cytotoxic activity, drugs are resuspended in medium containing lO~ fetal calf serum and incubated at 37O for l to WO97/18819 PCT~S96/18695 500 minutes. P388 cells are resuspended in the medium diluted to reach the appropriate drug concentrations. After 3 days incubation at 5~ CO2, cells are counted.
Induction of single strand breaks in P388 cells exposed to racemic fotemustine, ~+) fotemustine and ~ otemus~ine i5 compared. Cells (5XlO~) labelled 20 hours by (l4C)-thymidine are treated ~or 2 hours with various concentrations of ~otemustine and mixed with a similar number of (3H)-thymidine labelled reference cells. These cells are collected on 2.0 ~m pore size polycarbonate ~ilters and lysed with 5 mL
o~ 2~ SDS/0.025 M Na4EDTA ~pH9.7) in the presence o~
proteinase K. Elution is carried out in the dark at pH 12.l as described by Kohn. DNA single strand break ~requencies are calculated on the basis of ~irst order elution kinetics.
Induction of DNA-Protein cross-links is examined in P388 cells treated ~or various times in the presence of racemic fotemustine, (+) ~otemustine or ~-) fotemustine. (l4C)-Thymidine-labelled drug treated cells are irradiated with 600 rads o~ X-rays and (3H)-thymidine labelled reference cells are irradiated with 300 rads of X-rays. These cells are collected on 2.0~m pore-size polycarbonate ~ilters and lysed at pH 9.7 in the absence of proteinase K. Elution is the same as above.
Removal of DNA single strand breaks in ~ 388 cells at various times after exposure to concentrations of racemic fotemustine and its enantiomers causing equivalent DNA damage can also be _ CA 0223769~ 1998-0~-14 WO97/18819 PCT~S96/18695 m; n~d. The (l~C)-thymidine labelled cells initially exposed to drug treatment ~or 2 hours are washed and resuspended in drug- f ree medium. A~ter di~ferent times of incubation at 37~, these cel~s are mixed with an equal number of ~3H)-thymidine labelled reference cells, collected on the polycarbonate ~ilters and analyzed by alkaline elution.
The ~oregoing tests provide an estimate o~
relative activity, potency and selectivity.
Emesis in Ferrets. Experiments are performed on adult male ~errets. The ~im~l S are ~irst adapted to wearing a nylon jacket connected to a stainless-steel cable, which in turn is attached to a brass swivel at the cage top. After habituation to the tether-harness, each ~n;m~l receives a surgically implanted catheter in its right jugular vein. The catheter is ~lushed daily with heparinized sodium chloride. The drug studies are conducted l week a~ter the surgical procedure. Tethered animals are individually housed.
Eight to eleven ~nim~l s are used to evaluate each dose of each test compound. Individual animals are weighed weekly and randomly given, at greater than 48 hour intervals, a single i.v. or p.o. dose of racemic ~otemustine, (+)-~otemustine or (-)-~otemustine. At least three dose levels o~ each test compound are evaluated.
Individual animals are observed ~or 30 minutes ~ollowing administration o~ the test substance. The ~requency of, and latency to all expulsions, retches WO97/18819 PCT~S96/18C95 and de~ecations are recorded. Data obtained ~rom dose-response curves is tested ~or statistical significance by chi-square analysis. EDso values are determined ~or each compound. The test provides an estimate o~ relative liability to nausea and vomiting.
Claims (7)
1. A method of treating a malignancy which comprises administering to a mammal suffering from said malignancy, a therapeutically effective amount of (-)-fotemustine, substantially free of its (+) stereoisomer.
2. The method of claim 1 wherein said malignancy is chosen from the group consisting of lymphomas, melanomas, gliomas and other brain tumors, lung cancer, liver metastases and hematological and gastrointestinal malignancies.
3. The method of claim 1 wherein (-)-fotemustine is administered intravenously or intraarterially.
4. The method of claim 3 wherein the amount of (-)-fotemustine administered is from about 20 mg/m2 to about 150 mg/m2.
5. The method of claim 1 wherein the amount of (-)-fotemustine is greater than approximately 90% by weight of the total weight of fotemustine.
6. A pharmaceutical composition which comprises (-)-fotemustine substantially free of its (+) stereoisomer, and a pharmaceutically acceptable carrier.
7. The composition according to claim 6 adapted for parenteral delivery.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US736595P | 1995-11-20 | 1995-11-20 | |
| US60/007,365 | 1995-11-20 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2237695A1 true CA2237695A1 (en) | 1997-05-29 |
Family
ID=21725744
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002237695A Abandoned CA2237695A1 (en) | 1995-11-20 | 1996-11-19 | Antineoplastic methods and compositions employing optically pure (-)-fotemustine |
Country Status (5)
| Country | Link |
|---|---|
| EP (1) | EP0866712A4 (en) |
| JP (1) | JP2000500500A (en) |
| AU (1) | AU1057797A (en) |
| CA (1) | CA2237695A1 (en) |
| WO (1) | WO1997018819A1 (en) |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2536075B1 (en) * | 1982-11-17 | 1985-07-05 | Adir | NOVEL NITROSOUREE DERIVATIVES, THEIR PREPARATION PROCESS AND THE PHARMACEUTICAL COMPOSITIONS CONTAINING THEM |
-
1996
- 1996-11-19 WO PCT/US1996/018695 patent/WO1997018819A1/en not_active Application Discontinuation
- 1996-11-19 JP JP9519898A patent/JP2000500500A/en active Pending
- 1996-11-19 CA CA002237695A patent/CA2237695A1/en not_active Abandoned
- 1996-11-19 AU AU10577/97A patent/AU1057797A/en not_active Abandoned
- 1996-11-19 EP EP96941430A patent/EP0866712A4/en not_active Withdrawn
Also Published As
| Publication number | Publication date |
|---|---|
| EP0866712A4 (en) | 1998-12-23 |
| AU1057797A (en) | 1997-06-11 |
| EP0866712A1 (en) | 1998-09-30 |
| JP2000500500A (en) | 2000-01-18 |
| WO1997018819A1 (en) | 1997-05-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US5225404A (en) | Methods of treating colon tumors with tumor-inhibiting camptothecin compounds | |
| Schultz | Case study: high-dose intravenous vitamin C in the treatment of a patient with adenocarcinoma of the kidney | |
| KR0148589B1 (en) | Compositions, methods and kits for potentiating antitumor effect and for treating tumor | |
| BG98217A (en) | Determination of predecessors metabolized in the liver to biologically active products and their therapeutical application | |
| US20070036717A1 (en) | Chemoradiotherapy with TS-1/camptothecins | |
| JP2019528253A (en) | Radiohalogenating agents for in situ immunomodulatory cancer vaccination | |
| EA029072B1 (en) | Combination therapy comprising a dihydropyrazino-pyrazine compound and an androgen receptor antagonist for treating prostate cancer | |
| HU212102B (en) | Process for producing pharmaceutical composition for treating artherosclerosis containing pravastatin and captopril | |
| EP0344880B1 (en) | Pharmaceutical compositions with anti-cancer activity | |
| EP2301528B1 (en) | Use of FTS for treating malignant disorders | |
| JP2007063288A (en) | Method of treating cancer and pain associated therewith using endothelin antagonist | |
| US20210330626A1 (en) | Pharmaceutical composition for treating kidney cancer and application thereof | |
| US10463657B2 (en) | Method for treating osteoporosis, osteopenia or low bone mineral density | |
| EA007530B1 (en) | Antineoplastic combinations | |
| Friedman et al. | Increased melphalan activity in intracranial human medulloblastoma and glioma xenografts following buthionine sulfoximine-mediated glutathione depletion | |
| JP3437574B2 (en) | Use of creatine phosphate or phosphoenolpyruvate for the treatment of cancer | |
| JP2001521552A (en) | Combination therapy with chelerythrine | |
| Splinter et al. | Phase I study of alpha-difluoromethylornithine and methyl-GAG | |
| CA2237695A1 (en) | Antineoplastic methods and compositions employing optically pure (-)-fotemustine | |
| CN1151793C (en) | Medicinal compsns. for treating osseous lesion in multiple myeloma | |
| US20030143282A1 (en) | Adenosine A3 receptor agonist | |
| EP0868189A1 (en) | Antineoplastic methods and compositions employing (+)-fotemustine | |
| AU7229500A (en) | Antineoplastic methods and compositions employing optically pure (-)-fotemustine | |
| AU7229400A (en) | Antineoplastic methods and compositions employing (+)-fotemustine | |
| JP4846151B2 (en) | Use of treosulphan for conditioning patients prior to bone marrow or blood stem cell transplantation |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| FZDE | Dead |