CA2241255A1 - Method of treating disorders characterized by overexpression of cytidine deaminase or deoxycytidine deaminase - Google Patents
Method of treating disorders characterized by overexpression of cytidine deaminase or deoxycytidine deaminase Download PDFInfo
- Publication number
- CA2241255A1 CA2241255A1 CA002241255A CA2241255A CA2241255A1 CA 2241255 A1 CA2241255 A1 CA 2241255A1 CA 002241255 A CA002241255 A CA 002241255A CA 2241255 A CA2241255 A CA 2241255A CA 2241255 A1 CA2241255 A1 CA 2241255A1
- Authority
- CA
- Canada
- Prior art keywords
- agent
- composition
- group
- dideoxy
- azidocytidine
- 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
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
- A61K31/7052—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides
- A61K31/706—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom
-
- 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
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
-
- 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
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
- A61K31/7052—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides
- A61K31/706—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom
- A61K31/7064—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines
- A61K31/7068—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines having oxo groups directly attached to the pyrimidine ring, e.g. cytidine, cytidylic acid
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- 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
- A61P35/02—Antineoplastic agents specific for leukemia
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
- C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
- C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
- C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
- C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
- C07H19/06—Pyrimidine radicals
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
- C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
- C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
- C07H19/12—Triazine radicals
Abstract
A method of treating a disorder characterized by the overexpression of deaminase or deoxycytidine deaminase in a subject in need of such treatment is disclosed. The method comprises administering a compound of Formula (I) wherein X and X1 are each independently C or N; R1 is lower alkyl, lower alkenyl, lower alkynyl, halogen, or haloalkyl; and R2 is H, -N3, -OH, amino, or halogen; or a pharmaceutically acceptable salt thereof, to the subject in an amount effective to treat the disorder. Pharmaceutical formulations useful in the method of the present invention are also disclosed.
Description
, CA 02241255 1998-06-22 P~ 9 6 / 2 0 b4g ~P~/l~S 1 3 FEE3 1998 D--r/n,~9C/20649 AGENT ~ M~THOD FOR ~RE~TING DISORD~R8 ASSO~IATBD W~TH ~Yll~INE D~MTN~E OR
DEOXYCIT~DINE D~MTN~S~ O ~ R EX~RE~SION
~l~t~~ ~r~~ o~
This applicatlon i8 a continuation-in-part application of U. S. Patent A;pplication Serial No .
08/577 ,185 , f lled 22 D~c~mber 1995 .
-J R~r~r~o~ ND OF q~ ~v K ~ )N
Fiel ~ o~ the Tnver~ti or~
10The pre~ent inventlon r~latee to an active agent of the ~orlllula I and to treating a di80rder auQoclated inpar~ with the overexpress~on ~ cytidin~ ~e~mi na~e or deoxycytidine , ~e~mlnase~ comprising ~ni ~terin~ 5-methyl-2~, 3~-dideoxy-3'-azidocytidi.ne (5mAZC), analog thereof or a pharmaceutically ~ecti~e salt thereof to a ~i:ubj ect in need o~ -~uch treat~l3nt in an amo~mt e~ective to treat the disorder and ameliorat~ it~ ~ymptom~.
The overexpre~sion of cytidine de~min~e (CD) i~
~.~ associated with a number o~ human di.qorder~. For example, -J 20 certain kinds of leuke~ias are re~ractory to thc widely ~sed anti-cancer a~ene cytosine arabinoside (araC). The lack of response to araC ha~ been ~ound to be duc primarily either to inactivation o~ the deoxycytidine 3cinase gene locul: (who~e product i8 re~uired ;l~or activation ~ ar~C to its cancer-killing ~orm), or to the overexpression o~ cytidine de~min~~e or deoxycytidine deaminase (which deactivates araC by de~m;~ting it to an inactive i~orm, araUracil). There i8 cllrrently no adequate treatn~ent to overcome thi~ re~3iRt~3nce to arac dUQ to cytidine deamina~e or deoxycytidine ~e~;n~se ~rexpre~ion.
The overexpres~ion o~ cytidine de~m;n~e is al~o implicated in the in~ective cycle o~ the human YDE
~UB~___~ ~ r - . ~ CA 022412~ 1998-06-22 / 2 ~ 6 ~P~S I '~ ~ 8 2 PCT/US9 ~ ~
immunodeficiency virus (HIV), the virus implicated in AIDS. The initial states of HIV infection are characterized by the overexpression of cytidine de~m;n~e by the CD-4+T lymphocytes targeted by the virus. The elimination of this original set of infected cells could be critically important in controlling the level of subsequent infection.
Currently, one treatment for HIV infection and AIDS
is the administration of 3'-azido-3'deoxythymidine associated with this treatment. Problems with toxicity, however, have been associated with this treatment. Such toxicity occurs in part due to the fact that AZT is administered systemically, and damages host cells in all replicating tissue compartments.
There is currently a need for a treatment for HIV
infection that has the same or improved efficacy of AZT, without the side effects associated with toxicity. One solution to this problem would be a prodrug for AZT which exhibited reduced toxicity to replicating cells, and was preferentially activated to its virus-killing form (AZT) in HIV-infected cells.
In addition, inflammatory cells associated with the symptoms of arthritis have also been shown to overexpres~
'J cytidine de~m;n~se.
It would therefore be desirable to provide a relatively non-toxic prodrug that is preferentially activated to a metabolite that is toxic to arthritis mediating inflammatory cells.
SU~MA~y OF T~ lNv~:NllON
A method of treating a disorder associated with the overexpression of cytidine de~m-n~e or deoxycytidine de~m;n~e comprises administering to a subject in need of such treatment, of 5-methyl-2',3'dideoxy-3'azidocytidine (5mAZC), analog thereof or pharmaceutically acceptable salts thereof, hereinafter referred to as the "active compound", an amount effective to treat the disorder.
~ UIJ~lllU ~ " .''~V;~T
CA 022412~ 1998-06-22 9b~20 64 9 ~r~ ?'~ F~
DEOXYCIT~DINE D~MTN~S~ O ~ R EX~RE~SION
~l~t~~ ~r~~ o~
This applicatlon i8 a continuation-in-part application of U. S. Patent A;pplication Serial No .
08/577 ,185 , f lled 22 D~c~mber 1995 .
-J R~r~r~o~ ND OF q~ ~v K ~ )N
Fiel ~ o~ the Tnver~ti or~
10The pre~ent inventlon r~latee to an active agent of the ~orlllula I and to treating a di80rder auQoclated inpar~ with the overexpress~on ~ cytidin~ ~e~mi na~e or deoxycytidine , ~e~mlnase~ comprising ~ni ~terin~ 5-methyl-2~, 3~-dideoxy-3'-azidocytidi.ne (5mAZC), analog thereof or a pharmaceutically ~ecti~e salt thereof to a ~i:ubj ect in need o~ -~uch treat~l3nt in an amo~mt e~ective to treat the disorder and ameliorat~ it~ ~ymptom~.
The overexpre~sion of cytidine de~min~e (CD) i~
~.~ associated with a number o~ human di.qorder~. For example, -J 20 certain kinds of leuke~ias are re~ractory to thc widely ~sed anti-cancer a~ene cytosine arabinoside (araC). The lack of response to araC ha~ been ~ound to be duc primarily either to inactivation o~ the deoxycytidine 3cinase gene locul: (who~e product i8 re~uired ;l~or activation ~ ar~C to its cancer-killing ~orm), or to the overexpression o~ cytidine de~min~~e or deoxycytidine deaminase (which deactivates araC by de~m;~ting it to an inactive i~orm, araUracil). There i8 cllrrently no adequate treatn~ent to overcome thi~ re~3iRt~3nce to arac dUQ to cytidine deamina~e or deoxycytidine ~e~;n~se ~rexpre~ion.
The overexpres~ion o~ cytidine de~m;n~e is al~o implicated in the in~ective cycle o~ the human YDE
~UB~___~ ~ r - . ~ CA 022412~ 1998-06-22 / 2 ~ 6 ~P~S I '~ ~ 8 2 PCT/US9 ~ ~
immunodeficiency virus (HIV), the virus implicated in AIDS. The initial states of HIV infection are characterized by the overexpression of cytidine de~m;n~e by the CD-4+T lymphocytes targeted by the virus. The elimination of this original set of infected cells could be critically important in controlling the level of subsequent infection.
Currently, one treatment for HIV infection and AIDS
is the administration of 3'-azido-3'deoxythymidine associated with this treatment. Problems with toxicity, however, have been associated with this treatment. Such toxicity occurs in part due to the fact that AZT is administered systemically, and damages host cells in all replicating tissue compartments.
There is currently a need for a treatment for HIV
infection that has the same or improved efficacy of AZT, without the side effects associated with toxicity. One solution to this problem would be a prodrug for AZT which exhibited reduced toxicity to replicating cells, and was preferentially activated to its virus-killing form (AZT) in HIV-infected cells.
In addition, inflammatory cells associated with the symptoms of arthritis have also been shown to overexpres~
'J cytidine de~m;n~se.
It would therefore be desirable to provide a relatively non-toxic prodrug that is preferentially activated to a metabolite that is toxic to arthritis mediating inflammatory cells.
SU~MA~y OF T~ lNv~:NllON
A method of treating a disorder associated with the overexpression of cytidine de~m-n~e or deoxycytidine de~m;n~e comprises administering to a subject in need of such treatment, of 5-methyl-2',3'dideoxy-3'azidocytidine (5mAZC), analog thereof or pharmaceutically acceptable salts thereof, hereinafter referred to as the "active compound", an amount effective to treat the disorder.
~ UIJ~lllU ~ " .''~V;~T
CA 022412~ 1998-06-22 9b~20 64 9 ~r~ ?'~ F~
A method of treating a disorder associated with the overexpression of cytidine ~e~m;n~e or deoxycytidine d~m;n~.se comprises A~m;n;stering to a subject in need of the treatment, an amount of a compound of the chemical formula I
N~2 oD
H 2~' .
J
wherein X and Xlare each independently C o~ N;
R1 is lower alkyl, lower alkenyl, lower alkynyl, halogen, or haloalkyl; and R2is H, -N3 -OH, amino, or halogen;
or a pharmaceutically acceptable salt thereof, effective to treat the disorder.
A method of combating leukemia resistant to cytosine arabinoside (araC) comprises ~min;stering to a subject in need of the treatment an anti-araC resistant leukemia - effective amount of a compound of the chemical formula I.
A method of combating HIV-infection compris~s administering to a subject in need of the treatment an anti-HIV effective amount of a compound of the chemical formula I.
~ A method of combating arthritis comprises administering an anti-arthritis effective amount of a compound of the chemical formula I to an arthritic subject.
A method of combating a cancer associated with the overexpression of cytidine de~m;n~e, comprising administering to a subject in need of the treatment an AMEN~D SflEE~
;~ IJ ~S ~ _ ~, ~ .T
~ , .CA 02241255 1998-06-22 P~.,~ 9~2a~49 ? 3 FE~ ~9911 4 PCT/USs6/20649 ~nti-cancer e~~ective amount of a compo~nd o~ th~
chemical formula I.
The agent of this invention i8 suitable for the preparation o~ medicament for application in the treatment~ given above. The medicaments are provided in the form of oral, parenteral, topical, and transdermal formulations, a~ well a8 an implant and in the ~orm o~ a kit.
Rl;ITl~'~ n~ TpTTo~ OF ~ n~ ~
10Flgure 1 i3 a graph o~ a dose respon~e cur~e ~~ comparing the e~icacy of 5mAZC in HT-29SFC~ cells, which overexprer~.cytidine ~e~m;nA~e~ and HT-29SF cells, which do not. HT-29SF cell~ were plated in the bottom co~partment, and I~-29S~ cell~ in the top compartment of a "Transwell" culture dish ~l,OOO cells each). ~ A
semipermeable mem~rane separate~ the two compartments o~
the Tran~well, ~uch that any drug ~ n; ~tered in the ti88ue culture media ~athe both ~o~r~tmentn equally. The numbers in the x-axi8 indicate the concentration (in ~M) at which 5mAZC wa~ a~m~n1~tered continuou~ly or 72 hour~.
The num~ers on the y-axis indi~ate the number o~ ~urviving cell~. The circular (-) data polnts repre8ent the dose response for the HT-29SF''-n cells~. The ~quare ~) data point~ repre~ent the dose response ~or the ~T-29SF cell~.
25n~TAT~.~n nF~ ~N OF T~ ~Nv~l--~N
The method o~ the pre~ent invention may be ~cd to treat a di~order a-t~o~i~ted with the overexpression o~
cytidine deaminase and/or deoxycytidine de~m;n~e by administering an amount o~ the active compound o~ this invention to a subject in need o~ the treatment, e~fective to treat the di~order. That is 5-methyl 2',3'-deoxy-cytidine deAm;n~.qe(SmAZC) an analog t}l~reo~, or pharmaceutically acceptable ~alts thereo~ (i.e., the acti~e compound~). Examples o~ the disorde~ that may be treated include, but are not limited to, cancer, leukemia 'I~h~ ,f ~ S71~-Ç
_ P~ 6 ~ 2 0 649 ~P~f~S ' ~ FFB ~998 that i8 resistant to cytosine arabinoside (araC), HIV
infection, and arthritis. In a preferred embodiment, the method of the present invention is used to treat a subject suffering from a cancer that is characterized by the overexpression of cytidine ~e~min~e. Examples of these cancers include adenocarcinoma of the colon, adenocarcinoma of the lung, adenocarciona of the stomach, adenocarcinoma of the brea9t, Wilm's tumor, chondrocarcinoma, chondrosarcoma, leukemia, prostate tumors, brain tumors (e.g., glioma, astrocytoma), kidney tumors, pancreatic tumors, cervical tumors, liver tumors (e.g., hepatoblastoma, hepatocarcinoma), neuroblastoma, retinoblastoma, melanoma, basal cell carcinoma, sarcoma, and cancers metastatic to the liver (e.g., colon cancer).
While applicants do not wish to be bound to any particular theory of the instant invention, 5mAZC appears to be metabolized to 3'-azido-3'deoxythymidine (AZT) by cytidine de~m~nARe and deoxycytidine de~m;n~e, as illustrated in Scheme 1, below.
-NHI ' ll .CH~ CH~
o'J~N qn~ nc dC~ c N~
HOC~ HOC~I~
~_y~r~ y; ~ fi~r-Y-(~zn While not adversely affecting cells that do not overexpress cytidine de~mln~Rè, 5mAZC appears to be preferentially deaminated to AZT in tumor cells that overexpress cytidine de~mln~e.
The present invention is concerned primarily with the treatment of human subjects but may also be employed in A~ ID~
~ CA 02241255 1998-06-22 p~ 9 6 / ~D ~ ~ i ~ S ~3 FEBl998 the treatment of other m~mm~l ian subjects, such as dogs and cats, for veterinary purposes.
As used herein, the term "lower alkyl", refers to C
to C~ linear or branched alkyl, such as methyl, ethyl, propyl, butyl, isopropyl, sec-butyl, and tert-butyl.
Methyl is currently preferred. As used herein the term ~lower alkenyl", refers to c2 to C5 linear or br~nch~
alkenyl, such as ethenyl, propenyl, and butendyl. A8 sued herein, the term "lower alkynyl", refers to CZ to C5 linear or br~nche~ alykynl, such as propynyl and butynl. AS used herein, the term "haloalkyl" refers to a lower alkyl as defined above wherein one or more hydrogens is substituted -~ with a halo-group, e.g., a chloro-, fluor-, bromo- or iodo-group, with -CF3 being currently preferred.
Active compounds useful in the practice of the present invention include compounds of the chemical formula I
N~
ol~ ,X~ .
,.
wherein X and Xl are each independently C or N;
Rl is lower alkyl, lower alkenyl, -lower alkynyl, halogen, or haloalkyl; and R2 is H, -N3, -OH, amino, or halogen.
In a preferred embodiment of the present invention, Rl i8 methyl or -CF3, and R2 is -N3 or -OH. In a particularly preferred embodiment of the invention, Rl is methyl and R2 AMEND~SH~
- _ .CA 022412~ 1998-06-22 p~f~; 9 6 ~ 20 ~
IP~A/~; 13 FEB 19~
is -N3. Also encompassed herein are the pharmaceutically acceptable salts of these compounds.
Analogs of 5mAZC useful in the practice of the present invention include, but are not limited to, 5-methyl-2', 3'-dideoxycytidine, 5-ethyl-2',3'-dideoxycytidine, 5-ethyl-2', 3'-dideoxy-3'-azidocytidine, 5-propyl-2', 3'-di~eoxycytidine, 5-propyl-2', 3'-dideoxy-3'-azidocytidine, 5-propene-2',3'dideoxycytidine, 5 propene-2', 3'-dideoxy-3'-azidocytidine, 5-propyne-2',3'-dideoxycytidine, and 5-propyne-2',3'-dideoxy-3'-azidocytidine.
The structure of 5mAZC is known. See, e.g., T.S. ~in et al., J. Med. Chem 26: 1691-1696 (1983). 5mAZC is .-~available from ChemSyn Laboratories (Lenexa, Kansas, USA).
~15 Compounds useful for carrying out the present invention may be synthesized in accordance with known procedures which will be apparent to those skilled in the art. See, e.g., T.S. Lin et al., supra; T. Kulikowski et al., Acta Biochim. Polonica 16, 201-217 (1969); J.J. Fox et al., J.
Amer. Chem. Soc. 81, 178-187 (1959).
The active compounds disclosed herein may, as noted above, be prepared in the form of their pharmaceutically acceptable salts. "Pharmaceutically acceptable salts" are salts that retain the desired biological activity of the parent compound utilized and do not impart undesired - toxicological effects. Examples of such salts are (a) acid addition salts formed with inorganic acids, for example hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid and the like; and salts formed with organic acids such as, for example, acetic acid, oxalic acid, tartaric acid, succinic acid, maleic acid, fumaric acid, gluconic acid, citric acid, malic acid, ascorbic acid, benzoic acid, tannic acid, palmitic acid, alginic acid, polyglutamic acid, naphthalene sulfonic acid, methane sulfonic acid, p-toluenesulfonic acid, naphthalenedisulfonic acid, polygalacturonic acid, and the likei (b) salts formed from elemental anions such A~E~D~S~
~UIS:~lllUl~ .~
- CA 022412~ 1998-06-22 ~'~ 9 6 / 2 0 ~ '~
3 FEBI9~
as chlorine, bromine, and iodine, and (c) salts derived from bases, such as ammonium salts, alkali metal salts such as those of sodium and potassium, alkaline earth metal salts such as those of calcium and magnesium, and salts with organic bases such as dicyclohexylamine and N-methyl-D-glucamine.
The dosage of active compound for treatment will vary depending on the condition and the state of the subject being treated. Generally, the dosage may be as low as 0.1 ~mol/kg, but more preferably is at least 1.0 ~mol/kg and most preferably is at least 25 ~mol/kg. The dosage of the active compound may generally be as high as 1000 ~mol/kg, ~ more preferably is less than 500 ~mol/kg and still most preferably is less than 100 ~mol/kg. Depending on the solubility of the particular formulation of active compound administered, the daily dose may be divided among one or several unit dose administrations. The administration of the active compound of the invention may be carried out therapeutically, i.e., as a rescue treatment, or prophylactically.
Pharmaceutical compositions for use in the present method of treating disorders associated with the overexpression of cytidine d~m;n~se and/or deoxycytidine de~m;n~.~e include those suitable for inhalation, oral, rectal, parenteral (including subcutaneous, intradermal, intramuscular, intravenous) and transdermal adm~nistration. The compositions may conveniently be presented in unit dosage form and may be prepared by any of the methods which are well known in the art. The most suitable route of administration in any given case may depend upon the anatomic location of the disorder in the subject, the nature and severity of the condition being treated, and the particular formulation that is being used. The formulations may conveniently be presented in unit dosage form and may be prepared by any method well known in the art.
A~EN[~ S~
!~ U D:~ l l l IJ l l~
CA 0224125~ 1998-06-22 Pl~ 96/206 ; 3 FEE~ 199 9 PCT/usg6/2064s In the present method of treating leukemia or other disorders associated with the overexpression of cytidine de~m~n~qe or deoxycytidine deaminase, the active compound is administered in a dose range as given above. The dose of active agent may vary according to the condition being treated and the dose at which adverse pharmacological effects occur. One skilled in the art will take such factors into account when determining dosage.
In one embodiment of the present invention, the active compound is administered to a subject with araC-resistant leukemia in an amount effective so that it is metabolized by cytidine deAm-n~.~e or deoxycytidine '~ dPAm;n~Re to produce an anti-araC-resistant cancer e~fective amount o~ AZT.
15The active compound may be used alone or in combination with one or more anti-leukemic agents ~or the prophylaxis or treatment of araC-resistant leukemia.
In the manufacture of a pharmaceutical composition according to the invention into a "formulation", the active agent(s) or physiologically acceptable salts thereof (the "active compound") are typically admixed with, inter alia, an acceptable carrier. The carrier must be acceptable so that it is compatible with any other ~~, ingredients in the formulation and not deleterious to the '~ 25 subject's health in general. The carrier may be solid or liquid, or a mixture of both, and is preferably formulated wit~ the active compound as a unit-dose formulation, for example, a tablet, which may contain from 0.5~ to 99~ by weight of the active compound. One or more active compounds may be incorporated in the formulation of the invention, e.g., the formulation may contain one or more additional agen.s as noted above. The formulations may be prepared by any of the well known techniques of pharmacy comprising admixing the components with one or more carriers, and optionally including one or more accessory therapeutic ingredients.
A~EN~DS~E~
S~ Ll~ DE~T
CA 0224l2~ 1998-06-22 PGTf~ 9 b I ~ 4 ~
Formulations suitable for oral administration may be presented in discrete units, such as capsules, cachets, lozenges or tablets, each containing a predetermined amount of the active compound; as a powder or granules; as 5 a solution or a suspension in an aqueous or non-aqueous liquid; or as an oil-in-water or water-in-oil emulsion.
Such formulations may be prepared by any suitable method of pharmacy which includes bringing into association the active compound and a suitable carrier (which may contain 10 one or more accessory ingredients as noted above). In general, the formulations of the invention are prepared by uniformly and intimately admixing the active compound with ~ a liquid or finely divided solid carrier, or both, and J then, if necessary, shaping the resulting mixture. For 15 example, a tablet may be prepared by compressing or molding a powder or granules containing the active compound, optionally with one or more accessory ingredients. compressed tablets may be prepared by compressing, in a suitable machine, the compound in a 20 free-flowing form, such as a powder or granules optionally mixed with a binder, lubricant, inert diluent, and/or surface active/dispersing agent(s). Molded tablets may be made by molding in a suitable machine, the powdered compound moistened with an inert liquid binder. Liquid J 25 formulations for oral administration may optionally include enteric coatings known in the art to prevent degradation o~ the formulation in the stomach and provide release of the drug in the small intestine.
Formulations suitable for buccal (sub-lingual) 30 administration include lozenges comprising the active compound in a flavored base, usually sucrose and acacia or tragacanth; and pastilles comprising the compound in an inert base such as gelatin and glycerin or sucrose and acacla .
Formulations of the present invention suitable for parenteral administration comprise sterile aqueous and non-aqueous injection solutions of the active compound, AMENDED SH~
SUI~ ET
' ~ CA 022412~ 1998-06-22 ~.,~ 9 ~ ~ 20 ~ ~
S 1~ FEB 19 which prepara~ions are preferably isotonic with the blood of the intended recipient. These preparations may contain anti-oxidants, bu~ers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient. Aqueous and non-aqueous sterile suspensions may include suspending agents and thickening agents. The formulations may be presented in unit/dose or multi-dose containers, for example sealed capsules and vials, and may be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example, saline or water-for-injection immediately prior to use. Injection solutions and suspensions may be prepared from sterile powders, granules 3 and tablets o~ the kind previously described.
Formulations suitable for rectal administration are preferably presented as unit dose suppositories. These may be prepared by admixing the active compound wlth one or more conventional solid carriers, for example, cocoa butter, and then shaping the resulting mixture.
Formulations suitable for transdermal administration may be presented as discrete patches adapted to remain in intimate contact with the epidermis of the recipient ~or a prolonged period of time. Formulations suitable for transdermal administration may also be delivered by ~-~ 25 iontophoresis, and typically take the form of an optionally buffered aqueous solution of the active compound. See e.g., Pharmaceutical Research 3, 318 (1986).
The following examples are provided to more fully illustrate the present invention and should not be construed as restrictive thereof. In the following examples, g means grams, h means hours, kg means kilogram, ml means milliliter, M means molar, ~g means microgram, and ~mol means micromoles, nmol means nanomoles, pmol means picomoles, ~Ci means microCuries.
A~.~F~lD~ SHEFI
u11_ "ll~T
~ . CA 022412~ 1998-06-22 96~2~b'~
If~f~; 13 FEB t998 12 PCT/USg6/20649 ~XAMPT.~
~s~mP~e 1: Effect of 5mAZC in Ov6 6~re88ing Cytidine DeAm~nAae Cell~
HT-29SF human adenocarcinoma cells naturally express a high level of cytidine de~m;n~se (CD). HT-29SFCD is a subline of the HT-29SF line that express even higher CD
levels than normal (about 5.4 times the expression level of HT-29SF cells). In order to see whether cells that overexpress cytidine ~e~min~Re are more sensitive to 5mAZC
than other cells, HT-29SFC3 cells were plated in the top compartment of a Transwell culture dish, while HT-29SF
cells were placed in the bottom compartment (1000 cells each). A semipermeable membrane ~eparated the two compartments of the Transwell, such that drugs administered in the tissue culture media bathe both compartments eq~:a iy. 5mAZ~ was administered continuously for 72 hrs at concentrations ranging from 0 ~M to 50 ~M.
The results of this experiment are shown in Figure 1. The results of this experiment confirm the idea that cells that highly overexpress CD are sensitive to 5mAZC.
~Y~mDle ~: In Vitro Activity of 5mAZC
Based upon the results obtained in example 1 above, a dose of 50 ~M 5mAZC was selected for more in-depth analysis. Two experiments were performed, in quintuplicate, in the same Transwell dishes, containing the HT-29SF cells and the HT-29SFCD cells in compartments separated only by a membrane permeable to 5mAZC. The results are given in Table 1 below.
l l l U l ~ r- l ~ CA 0224l2~ l998-06-22 - Pt T~ 9 6 ~
~pFA~S 1~ FEB 1998 Table 1. E~ects of exposure o~ HT-29SF and HF-29SFCD human colonic adenocarcinoma cells to 50~M 5mAZC (72h) in Transwell culture system dishes.
Experiment 1 Example 2 Number of viable cells Number of viable cells 33. 8 i: 17.1 797i 68 45.4 :: 15.6 867.2 i 29.4 le ~ In vivo Activity of 5mAZC:
~ n Tumor Xenograft Model To demonstrate that 5mAZC exhibits in vivo activity, ;~ two sixteen week old female Balb/C nu/nu mice were subcutaneously inoculated with 2.0 x 106 HT-29SF (non cytidine d~m,n~e producing)cells in the le~t scapular region, and 2.0 x 106 HT-29SFCD (cytidine d~m;n~e producing)cells in the right scapular region. One of these mice received twice daily intraperitoneal doses of 150 mg/kg of 5mAZC. The other mouse received saline only.
This treatment continued for 5 weeks, at which time the animals were euthanized and the solid tumors were dissected free and weighed. The ~Am;ned tumors had the following weights:
) 1- HT-29SFCD + 5mAZC: 0.22 g i 0.31 g (N=2) 2- HT-29SF + 5mAZC: 1.02 _ 0.13 g (N=2) 3- HT-29SFCD + saline: 0.96 g (N=l) 4- HT-29SF + saline: 1.13 g (N=l) In another experiment, thirty BALB/c nude mice received subcutaneous injections of 5 x 106 HT-29SF cells.
Twelve of the animals received 600 mg/kg/day 5mAZC in the flank for 28 days. Eighteen animals received saline injections on the same schedule. On Day 29, the animals were sacrificed and the tumor volume was quantified by standard methods. The average tumor volume for the control group receiving saline was 509 ~ 292 mm3. In contrast, the animals treated with 5mAZC had an average tumor volume of 273 + 97 mm3, an average 46~ reduction in ~MENDEO SHEET
~nRs~lTUT~ L~
CA 0224l2~ l998-06-22 r; ~ ? .~.
14 IP ~ ~ /U3936~ ~ ~398 size. The results of Examples 2 and 3 show clearly that 5mAZC is activated in vitro and in vivo into molecular species exclusively toxic to tumor cells that overexpress cytidine deaminase.
~Y~ple g: CD Overexpression in Colon Tumora:
Ex vivo Patient Data The hypothesis that cytidine de~m;n~e (CD) i8 overexpressed in certain tumors as compared with normal tissue was demonstrated in experiments conducted with surgical specimens obtained from patient undergoing bowel resection for colon cancer. Specimens of tumor and adjacent normal tissue obtained from pathological evaluation ere assayed for CD activity using [3H]-cytidine as a substrate, according to the method of R.L. Momparler and J. Laliberte, J. Leukemia Res., 14 (9), 751-54 (1990).
Briefly, colon tumors and adjacent mucosa from surgical specimens were homogenized in 5mM Tris-C1, pH 7.4, sonicated on ice in three 5-second pulses, and then adjusted to 50mM Tris-HCl,pH 7.4. The homogenates were centrifuged at 12,000 x g for 15 minutes at 4~C, and the homogenates stored at -70~C until analysis. Protein concentration was determined using a BioRad protein assay , .~, J kit and bovine gamma globulin was used as a standard.
For determination of cytidine de~m;n~e activity, the reaction mixture (100~L) contained 25 mM Tris-Cl, pH 7.4, 0.5 ~Ci of [3H]-cytidine and 0.02-0.05 mg of homogenate tissue. Reactions were carried out at 37~C for 30 minutes, then stopped with 3 mL of cold 0.001 N HCl. The reaction mixture was placed on Whatman P-81 phosphocellulose discs that were washed with 3 mL of H2O, 1 mL of 0.1 N HC1, and 3 mL of H20 twice before use. The mixture was allowed to flow gently by gravity for 1 hr and then assayed for radioactivity. The results were normalized for protein content and expressed as nmol/min/mg protein. The results of this experiment are presented in Table 2 below.
AME~'IDED SH~E~
CA 0224l2~ l998-06-22 ~xample 5 Preferential Deamination of 5mAZC to AZT in Colonic Tumors:
The hypothesis that 5mAZC is preferentially deaminated to AZT in human colonic tumors versus in normal adjacent mucosa was demonstrated in the same ex vivo samples described above in Example 4. Using ~3H]-5mAZC as a substrate, the ~3H]-AZT product of the enzymatic reaction was isolated by HPLC and the radiolabeled product quantitated as described in J.W.
Nyce et al., Proc. Natl . Acad. Sci USA 90 , 2960-2964 (1993). Briefly, procedures described in Example 4 were repeated with the following exceptions: Each reaction mixture contained 0.26 ~Ci o~ [3H]-5mAZC. The reactions were terminated with 5~ ~L of 2M perchloric acid and the mixture separated by HPLC using a 4.6 x 25 cm Beckman Ultrasphere ODS with a mobile phase o~ 12.5~ acetonitrile in 40 mM sodium acetate pH 7Ø Unlabeled 5mAZC and AZT
were used to authenticate the resulting peaks. The fractions corresponding to radiolabeled 5mAZC and AZT
were collected and assayed for radioactivity by scintillation counting. The results were normalized for protein content and expressed as pmol/min/mg protein, and are provided below in Table 3.
To confirm that the AZT found in the colonic tumor tissue was being produced by cytidine d~m;n~Re activity (see Scheme 1, above), the cytidine de~m;n~Re inhibitor tetrahydrouridine (THU) was added to the reaction mixtures containing tumor tissue homogenate and radiolabeled 5mAZC; the amount of radiolabeled AZT
produced was assayed as above, then compared to the amount of radiolabeled A~T produced in reaction mixtures not containing THU. The results of this experiment are also provided below in Table 3.
The foregoing Examples are illustrative of the present invention, and are not to be construed as limiting thereof. The invention is defined by the CA 022412~ 1998-06-22 W097~3230 PCT~S96/20649 following claims, with equivalents o~ the claims to be included therein.
Ex Vivo Patient Data:
5Cytidine Deaminase Activity in Tumor vs. Normal Adjacent Mucosa PatientActivity In Normal Activity inPercentage p value Mucosa AdjacentAdenocarcimoma Tumor CD
to Tumor Tumor Activity o~
nmol/min/mg nmol/min/mg Normal protein protein 13.26 i 0.67 17.36 i 0.24 130.92 < .01 212.00 i 0.50 15.49 i 0.33 129.08 <.01 10 3 8.81 i 0.79 29.20 i 0.54 331.44 <.01 410.82 i 0.63 12.98 i 0.58 119.96 < .01 18.38 i 0.34 169.87 5 7.78 i 0.22 9.25 i 0.48 118.89 < .05 617.32 i 0.93 22.35 ~ 0.39 129.04 < .01 7 6.37 i 0.89 18.69 ~ 1.27 294.98 < .01 15 8 9.63 i 0.77 14.55 i 1.17 151.09 <.05 914.01 i 0.89 20.09 i 0.20 143.40 < .01 10 6.06 i 0.83 18.51 i 0.89 305.22 <.01 Average10.61 i 3.6 17.91 i 5.2 184 HT-29SF 11.96 i 0.16 CA 0224l2~ l998-06-22 WO 97/23230 PCT/US96/~0649 T~BLE 3 Ex vivo patient data:
Preferential 5mAZC deamination to AZT in tumor vs. normal adjacent mucosa 5 Patient # Radiolabeled AZT Radiolabeled AZT Percentage p value in Normal in Colon TumorAZT in Tumor AdjacentTissue Tissue Tissue of pmol/min/mg pmol/min/mg Normal protein protein 117.14 i 7.61 134.51 i 14.92 114.83 After THU:
30.56 i 4.22 2 128.76 i 2.17 153.34 i 8.70 119.09 < .05 After THU:
50.48 i 7.30 3 40.42 i 1.18304.04 i 17.63 752.17 < .01 After THU:
0.31 i 0.31 4 77.52 i 8.8380.01 i 20.23 103.33 polyp After THU:
N~2 oD
H 2~' .
J
wherein X and Xlare each independently C o~ N;
R1 is lower alkyl, lower alkenyl, lower alkynyl, halogen, or haloalkyl; and R2is H, -N3 -OH, amino, or halogen;
or a pharmaceutically acceptable salt thereof, effective to treat the disorder.
A method of combating leukemia resistant to cytosine arabinoside (araC) comprises ~min;stering to a subject in need of the treatment an anti-araC resistant leukemia - effective amount of a compound of the chemical formula I.
A method of combating HIV-infection compris~s administering to a subject in need of the treatment an anti-HIV effective amount of a compound of the chemical formula I.
~ A method of combating arthritis comprises administering an anti-arthritis effective amount of a compound of the chemical formula I to an arthritic subject.
A method of combating a cancer associated with the overexpression of cytidine de~m;n~e, comprising administering to a subject in need of the treatment an AMEN~D SflEE~
;~ IJ ~S ~ _ ~, ~ .T
~ , .CA 02241255 1998-06-22 P~.,~ 9~2a~49 ? 3 FE~ ~9911 4 PCT/USs6/20649 ~nti-cancer e~~ective amount of a compo~nd o~ th~
chemical formula I.
The agent of this invention i8 suitable for the preparation o~ medicament for application in the treatment~ given above. The medicaments are provided in the form of oral, parenteral, topical, and transdermal formulations, a~ well a8 an implant and in the ~orm o~ a kit.
Rl;ITl~'~ n~ TpTTo~ OF ~ n~ ~
10Flgure 1 i3 a graph o~ a dose respon~e cur~e ~~ comparing the e~icacy of 5mAZC in HT-29SFC~ cells, which overexprer~.cytidine ~e~m;nA~e~ and HT-29SF cells, which do not. HT-29SF cell~ were plated in the bottom co~partment, and I~-29S~ cell~ in the top compartment of a "Transwell" culture dish ~l,OOO cells each). ~ A
semipermeable mem~rane separate~ the two compartments o~
the Tran~well, ~uch that any drug ~ n; ~tered in the ti88ue culture media ~athe both ~o~r~tmentn equally. The numbers in the x-axi8 indicate the concentration (in ~M) at which 5mAZC wa~ a~m~n1~tered continuou~ly or 72 hour~.
The num~ers on the y-axis indi~ate the number o~ ~urviving cell~. The circular (-) data polnts repre8ent the dose response for the HT-29SF''-n cells~. The ~quare ~) data point~ repre~ent the dose response ~or the ~T-29SF cell~.
25n~TAT~.~n nF~ ~N OF T~ ~Nv~l--~N
The method o~ the pre~ent invention may be ~cd to treat a di~order a-t~o~i~ted with the overexpression o~
cytidine deaminase and/or deoxycytidine de~m;n~e by administering an amount o~ the active compound o~ this invention to a subject in need o~ the treatment, e~fective to treat the di~order. That is 5-methyl 2',3'-deoxy-cytidine deAm;n~.qe(SmAZC) an analog t}l~reo~, or pharmaceutically acceptable ~alts thereo~ (i.e., the acti~e compound~). Examples o~ the disorde~ that may be treated include, but are not limited to, cancer, leukemia 'I~h~ ,f ~ S71~-Ç
_ P~ 6 ~ 2 0 649 ~P~f~S ' ~ FFB ~998 that i8 resistant to cytosine arabinoside (araC), HIV
infection, and arthritis. In a preferred embodiment, the method of the present invention is used to treat a subject suffering from a cancer that is characterized by the overexpression of cytidine ~e~min~e. Examples of these cancers include adenocarcinoma of the colon, adenocarcinoma of the lung, adenocarciona of the stomach, adenocarcinoma of the brea9t, Wilm's tumor, chondrocarcinoma, chondrosarcoma, leukemia, prostate tumors, brain tumors (e.g., glioma, astrocytoma), kidney tumors, pancreatic tumors, cervical tumors, liver tumors (e.g., hepatoblastoma, hepatocarcinoma), neuroblastoma, retinoblastoma, melanoma, basal cell carcinoma, sarcoma, and cancers metastatic to the liver (e.g., colon cancer).
While applicants do not wish to be bound to any particular theory of the instant invention, 5mAZC appears to be metabolized to 3'-azido-3'deoxythymidine (AZT) by cytidine de~m~nARe and deoxycytidine de~m;n~e, as illustrated in Scheme 1, below.
-NHI ' ll .CH~ CH~
o'J~N qn~ nc dC~ c N~
HOC~ HOC~I~
~_y~r~ y; ~ fi~r-Y-(~zn While not adversely affecting cells that do not overexpress cytidine de~mln~Rè, 5mAZC appears to be preferentially deaminated to AZT in tumor cells that overexpress cytidine de~mln~e.
The present invention is concerned primarily with the treatment of human subjects but may also be employed in A~ ID~
~ CA 02241255 1998-06-22 p~ 9 6 / ~D ~ ~ i ~ S ~3 FEBl998 the treatment of other m~mm~l ian subjects, such as dogs and cats, for veterinary purposes.
As used herein, the term "lower alkyl", refers to C
to C~ linear or branched alkyl, such as methyl, ethyl, propyl, butyl, isopropyl, sec-butyl, and tert-butyl.
Methyl is currently preferred. As used herein the term ~lower alkenyl", refers to c2 to C5 linear or br~nch~
alkenyl, such as ethenyl, propenyl, and butendyl. A8 sued herein, the term "lower alkynyl", refers to CZ to C5 linear or br~nche~ alykynl, such as propynyl and butynl. AS used herein, the term "haloalkyl" refers to a lower alkyl as defined above wherein one or more hydrogens is substituted -~ with a halo-group, e.g., a chloro-, fluor-, bromo- or iodo-group, with -CF3 being currently preferred.
Active compounds useful in the practice of the present invention include compounds of the chemical formula I
N~
ol~ ,X~ .
,.
wherein X and Xl are each independently C or N;
Rl is lower alkyl, lower alkenyl, -lower alkynyl, halogen, or haloalkyl; and R2 is H, -N3, -OH, amino, or halogen.
In a preferred embodiment of the present invention, Rl i8 methyl or -CF3, and R2 is -N3 or -OH. In a particularly preferred embodiment of the invention, Rl is methyl and R2 AMEND~SH~
- _ .CA 022412~ 1998-06-22 p~f~; 9 6 ~ 20 ~
IP~A/~; 13 FEB 19~
is -N3. Also encompassed herein are the pharmaceutically acceptable salts of these compounds.
Analogs of 5mAZC useful in the practice of the present invention include, but are not limited to, 5-methyl-2', 3'-dideoxycytidine, 5-ethyl-2',3'-dideoxycytidine, 5-ethyl-2', 3'-dideoxy-3'-azidocytidine, 5-propyl-2', 3'-di~eoxycytidine, 5-propyl-2', 3'-dideoxy-3'-azidocytidine, 5-propene-2',3'dideoxycytidine, 5 propene-2', 3'-dideoxy-3'-azidocytidine, 5-propyne-2',3'-dideoxycytidine, and 5-propyne-2',3'-dideoxy-3'-azidocytidine.
The structure of 5mAZC is known. See, e.g., T.S. ~in et al., J. Med. Chem 26: 1691-1696 (1983). 5mAZC is .-~available from ChemSyn Laboratories (Lenexa, Kansas, USA).
~15 Compounds useful for carrying out the present invention may be synthesized in accordance with known procedures which will be apparent to those skilled in the art. See, e.g., T.S. Lin et al., supra; T. Kulikowski et al., Acta Biochim. Polonica 16, 201-217 (1969); J.J. Fox et al., J.
Amer. Chem. Soc. 81, 178-187 (1959).
The active compounds disclosed herein may, as noted above, be prepared in the form of their pharmaceutically acceptable salts. "Pharmaceutically acceptable salts" are salts that retain the desired biological activity of the parent compound utilized and do not impart undesired - toxicological effects. Examples of such salts are (a) acid addition salts formed with inorganic acids, for example hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid and the like; and salts formed with organic acids such as, for example, acetic acid, oxalic acid, tartaric acid, succinic acid, maleic acid, fumaric acid, gluconic acid, citric acid, malic acid, ascorbic acid, benzoic acid, tannic acid, palmitic acid, alginic acid, polyglutamic acid, naphthalene sulfonic acid, methane sulfonic acid, p-toluenesulfonic acid, naphthalenedisulfonic acid, polygalacturonic acid, and the likei (b) salts formed from elemental anions such A~E~D~S~
~UIS:~lllUl~ .~
- CA 022412~ 1998-06-22 ~'~ 9 6 / 2 0 ~ '~
3 FEBI9~
as chlorine, bromine, and iodine, and (c) salts derived from bases, such as ammonium salts, alkali metal salts such as those of sodium and potassium, alkaline earth metal salts such as those of calcium and magnesium, and salts with organic bases such as dicyclohexylamine and N-methyl-D-glucamine.
The dosage of active compound for treatment will vary depending on the condition and the state of the subject being treated. Generally, the dosage may be as low as 0.1 ~mol/kg, but more preferably is at least 1.0 ~mol/kg and most preferably is at least 25 ~mol/kg. The dosage of the active compound may generally be as high as 1000 ~mol/kg, ~ more preferably is less than 500 ~mol/kg and still most preferably is less than 100 ~mol/kg. Depending on the solubility of the particular formulation of active compound administered, the daily dose may be divided among one or several unit dose administrations. The administration of the active compound of the invention may be carried out therapeutically, i.e., as a rescue treatment, or prophylactically.
Pharmaceutical compositions for use in the present method of treating disorders associated with the overexpression of cytidine d~m;n~se and/or deoxycytidine de~m;n~.~e include those suitable for inhalation, oral, rectal, parenteral (including subcutaneous, intradermal, intramuscular, intravenous) and transdermal adm~nistration. The compositions may conveniently be presented in unit dosage form and may be prepared by any of the methods which are well known in the art. The most suitable route of administration in any given case may depend upon the anatomic location of the disorder in the subject, the nature and severity of the condition being treated, and the particular formulation that is being used. The formulations may conveniently be presented in unit dosage form and may be prepared by any method well known in the art.
A~EN[~ S~
!~ U D:~ l l l IJ l l~
CA 0224125~ 1998-06-22 Pl~ 96/206 ; 3 FEE~ 199 9 PCT/usg6/2064s In the present method of treating leukemia or other disorders associated with the overexpression of cytidine de~m~n~qe or deoxycytidine deaminase, the active compound is administered in a dose range as given above. The dose of active agent may vary according to the condition being treated and the dose at which adverse pharmacological effects occur. One skilled in the art will take such factors into account when determining dosage.
In one embodiment of the present invention, the active compound is administered to a subject with araC-resistant leukemia in an amount effective so that it is metabolized by cytidine deAm-n~.~e or deoxycytidine '~ dPAm;n~Re to produce an anti-araC-resistant cancer e~fective amount o~ AZT.
15The active compound may be used alone or in combination with one or more anti-leukemic agents ~or the prophylaxis or treatment of araC-resistant leukemia.
In the manufacture of a pharmaceutical composition according to the invention into a "formulation", the active agent(s) or physiologically acceptable salts thereof (the "active compound") are typically admixed with, inter alia, an acceptable carrier. The carrier must be acceptable so that it is compatible with any other ~~, ingredients in the formulation and not deleterious to the '~ 25 subject's health in general. The carrier may be solid or liquid, or a mixture of both, and is preferably formulated wit~ the active compound as a unit-dose formulation, for example, a tablet, which may contain from 0.5~ to 99~ by weight of the active compound. One or more active compounds may be incorporated in the formulation of the invention, e.g., the formulation may contain one or more additional agen.s as noted above. The formulations may be prepared by any of the well known techniques of pharmacy comprising admixing the components with one or more carriers, and optionally including one or more accessory therapeutic ingredients.
A~EN~DS~E~
S~ Ll~ DE~T
CA 0224l2~ 1998-06-22 PGTf~ 9 b I ~ 4 ~
Formulations suitable for oral administration may be presented in discrete units, such as capsules, cachets, lozenges or tablets, each containing a predetermined amount of the active compound; as a powder or granules; as 5 a solution or a suspension in an aqueous or non-aqueous liquid; or as an oil-in-water or water-in-oil emulsion.
Such formulations may be prepared by any suitable method of pharmacy which includes bringing into association the active compound and a suitable carrier (which may contain 10 one or more accessory ingredients as noted above). In general, the formulations of the invention are prepared by uniformly and intimately admixing the active compound with ~ a liquid or finely divided solid carrier, or both, and J then, if necessary, shaping the resulting mixture. For 15 example, a tablet may be prepared by compressing or molding a powder or granules containing the active compound, optionally with one or more accessory ingredients. compressed tablets may be prepared by compressing, in a suitable machine, the compound in a 20 free-flowing form, such as a powder or granules optionally mixed with a binder, lubricant, inert diluent, and/or surface active/dispersing agent(s). Molded tablets may be made by molding in a suitable machine, the powdered compound moistened with an inert liquid binder. Liquid J 25 formulations for oral administration may optionally include enteric coatings known in the art to prevent degradation o~ the formulation in the stomach and provide release of the drug in the small intestine.
Formulations suitable for buccal (sub-lingual) 30 administration include lozenges comprising the active compound in a flavored base, usually sucrose and acacia or tragacanth; and pastilles comprising the compound in an inert base such as gelatin and glycerin or sucrose and acacla .
Formulations of the present invention suitable for parenteral administration comprise sterile aqueous and non-aqueous injection solutions of the active compound, AMENDED SH~
SUI~ ET
' ~ CA 022412~ 1998-06-22 ~.,~ 9 ~ ~ 20 ~ ~
S 1~ FEB 19 which prepara~ions are preferably isotonic with the blood of the intended recipient. These preparations may contain anti-oxidants, bu~ers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient. Aqueous and non-aqueous sterile suspensions may include suspending agents and thickening agents. The formulations may be presented in unit/dose or multi-dose containers, for example sealed capsules and vials, and may be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example, saline or water-for-injection immediately prior to use. Injection solutions and suspensions may be prepared from sterile powders, granules 3 and tablets o~ the kind previously described.
Formulations suitable for rectal administration are preferably presented as unit dose suppositories. These may be prepared by admixing the active compound wlth one or more conventional solid carriers, for example, cocoa butter, and then shaping the resulting mixture.
Formulations suitable for transdermal administration may be presented as discrete patches adapted to remain in intimate contact with the epidermis of the recipient ~or a prolonged period of time. Formulations suitable for transdermal administration may also be delivered by ~-~ 25 iontophoresis, and typically take the form of an optionally buffered aqueous solution of the active compound. See e.g., Pharmaceutical Research 3, 318 (1986).
The following examples are provided to more fully illustrate the present invention and should not be construed as restrictive thereof. In the following examples, g means grams, h means hours, kg means kilogram, ml means milliliter, M means molar, ~g means microgram, and ~mol means micromoles, nmol means nanomoles, pmol means picomoles, ~Ci means microCuries.
A~.~F~lD~ SHEFI
u11_ "ll~T
~ . CA 022412~ 1998-06-22 96~2~b'~
If~f~; 13 FEB t998 12 PCT/USg6/20649 ~XAMPT.~
~s~mP~e 1: Effect of 5mAZC in Ov6 6~re88ing Cytidine DeAm~nAae Cell~
HT-29SF human adenocarcinoma cells naturally express a high level of cytidine de~m;n~se (CD). HT-29SFCD is a subline of the HT-29SF line that express even higher CD
levels than normal (about 5.4 times the expression level of HT-29SF cells). In order to see whether cells that overexpress cytidine ~e~min~Re are more sensitive to 5mAZC
than other cells, HT-29SFC3 cells were plated in the top compartment of a Transwell culture dish, while HT-29SF
cells were placed in the bottom compartment (1000 cells each). A semipermeable membrane ~eparated the two compartments of the Transwell, such that drugs administered in the tissue culture media bathe both compartments eq~:a iy. 5mAZ~ was administered continuously for 72 hrs at concentrations ranging from 0 ~M to 50 ~M.
The results of this experiment are shown in Figure 1. The results of this experiment confirm the idea that cells that highly overexpress CD are sensitive to 5mAZC.
~Y~mDle ~: In Vitro Activity of 5mAZC
Based upon the results obtained in example 1 above, a dose of 50 ~M 5mAZC was selected for more in-depth analysis. Two experiments were performed, in quintuplicate, in the same Transwell dishes, containing the HT-29SF cells and the HT-29SFCD cells in compartments separated only by a membrane permeable to 5mAZC. The results are given in Table 1 below.
l l l U l ~ r- l ~ CA 0224l2~ l998-06-22 - Pt T~ 9 6 ~
~pFA~S 1~ FEB 1998 Table 1. E~ects of exposure o~ HT-29SF and HF-29SFCD human colonic adenocarcinoma cells to 50~M 5mAZC (72h) in Transwell culture system dishes.
Experiment 1 Example 2 Number of viable cells Number of viable cells 33. 8 i: 17.1 797i 68 45.4 :: 15.6 867.2 i 29.4 le ~ In vivo Activity of 5mAZC:
~ n Tumor Xenograft Model To demonstrate that 5mAZC exhibits in vivo activity, ;~ two sixteen week old female Balb/C nu/nu mice were subcutaneously inoculated with 2.0 x 106 HT-29SF (non cytidine d~m,n~e producing)cells in the le~t scapular region, and 2.0 x 106 HT-29SFCD (cytidine d~m;n~e producing)cells in the right scapular region. One of these mice received twice daily intraperitoneal doses of 150 mg/kg of 5mAZC. The other mouse received saline only.
This treatment continued for 5 weeks, at which time the animals were euthanized and the solid tumors were dissected free and weighed. The ~Am;ned tumors had the following weights:
) 1- HT-29SFCD + 5mAZC: 0.22 g i 0.31 g (N=2) 2- HT-29SF + 5mAZC: 1.02 _ 0.13 g (N=2) 3- HT-29SFCD + saline: 0.96 g (N=l) 4- HT-29SF + saline: 1.13 g (N=l) In another experiment, thirty BALB/c nude mice received subcutaneous injections of 5 x 106 HT-29SF cells.
Twelve of the animals received 600 mg/kg/day 5mAZC in the flank for 28 days. Eighteen animals received saline injections on the same schedule. On Day 29, the animals were sacrificed and the tumor volume was quantified by standard methods. The average tumor volume for the control group receiving saline was 509 ~ 292 mm3. In contrast, the animals treated with 5mAZC had an average tumor volume of 273 + 97 mm3, an average 46~ reduction in ~MENDEO SHEET
~nRs~lTUT~ L~
CA 0224l2~ l998-06-22 r; ~ ? .~.
14 IP ~ ~ /U3936~ ~ ~398 size. The results of Examples 2 and 3 show clearly that 5mAZC is activated in vitro and in vivo into molecular species exclusively toxic to tumor cells that overexpress cytidine deaminase.
~Y~ple g: CD Overexpression in Colon Tumora:
Ex vivo Patient Data The hypothesis that cytidine de~m;n~e (CD) i8 overexpressed in certain tumors as compared with normal tissue was demonstrated in experiments conducted with surgical specimens obtained from patient undergoing bowel resection for colon cancer. Specimens of tumor and adjacent normal tissue obtained from pathological evaluation ere assayed for CD activity using [3H]-cytidine as a substrate, according to the method of R.L. Momparler and J. Laliberte, J. Leukemia Res., 14 (9), 751-54 (1990).
Briefly, colon tumors and adjacent mucosa from surgical specimens were homogenized in 5mM Tris-C1, pH 7.4, sonicated on ice in three 5-second pulses, and then adjusted to 50mM Tris-HCl,pH 7.4. The homogenates were centrifuged at 12,000 x g for 15 minutes at 4~C, and the homogenates stored at -70~C until analysis. Protein concentration was determined using a BioRad protein assay , .~, J kit and bovine gamma globulin was used as a standard.
For determination of cytidine de~m;n~e activity, the reaction mixture (100~L) contained 25 mM Tris-Cl, pH 7.4, 0.5 ~Ci of [3H]-cytidine and 0.02-0.05 mg of homogenate tissue. Reactions were carried out at 37~C for 30 minutes, then stopped with 3 mL of cold 0.001 N HCl. The reaction mixture was placed on Whatman P-81 phosphocellulose discs that were washed with 3 mL of H2O, 1 mL of 0.1 N HC1, and 3 mL of H20 twice before use. The mixture was allowed to flow gently by gravity for 1 hr and then assayed for radioactivity. The results were normalized for protein content and expressed as nmol/min/mg protein. The results of this experiment are presented in Table 2 below.
AME~'IDED SH~E~
CA 0224l2~ l998-06-22 ~xample 5 Preferential Deamination of 5mAZC to AZT in Colonic Tumors:
The hypothesis that 5mAZC is preferentially deaminated to AZT in human colonic tumors versus in normal adjacent mucosa was demonstrated in the same ex vivo samples described above in Example 4. Using ~3H]-5mAZC as a substrate, the ~3H]-AZT product of the enzymatic reaction was isolated by HPLC and the radiolabeled product quantitated as described in J.W.
Nyce et al., Proc. Natl . Acad. Sci USA 90 , 2960-2964 (1993). Briefly, procedures described in Example 4 were repeated with the following exceptions: Each reaction mixture contained 0.26 ~Ci o~ [3H]-5mAZC. The reactions were terminated with 5~ ~L of 2M perchloric acid and the mixture separated by HPLC using a 4.6 x 25 cm Beckman Ultrasphere ODS with a mobile phase o~ 12.5~ acetonitrile in 40 mM sodium acetate pH 7Ø Unlabeled 5mAZC and AZT
were used to authenticate the resulting peaks. The fractions corresponding to radiolabeled 5mAZC and AZT
were collected and assayed for radioactivity by scintillation counting. The results were normalized for protein content and expressed as pmol/min/mg protein, and are provided below in Table 3.
To confirm that the AZT found in the colonic tumor tissue was being produced by cytidine d~m;n~Re activity (see Scheme 1, above), the cytidine de~m;n~Re inhibitor tetrahydrouridine (THU) was added to the reaction mixtures containing tumor tissue homogenate and radiolabeled 5mAZC; the amount of radiolabeled AZT
produced was assayed as above, then compared to the amount of radiolabeled A~T produced in reaction mixtures not containing THU. The results of this experiment are also provided below in Table 3.
The foregoing Examples are illustrative of the present invention, and are not to be construed as limiting thereof. The invention is defined by the CA 022412~ 1998-06-22 W097~3230 PCT~S96/20649 following claims, with equivalents o~ the claims to be included therein.
Ex Vivo Patient Data:
5Cytidine Deaminase Activity in Tumor vs. Normal Adjacent Mucosa PatientActivity In Normal Activity inPercentage p value Mucosa AdjacentAdenocarcimoma Tumor CD
to Tumor Tumor Activity o~
nmol/min/mg nmol/min/mg Normal protein protein 13.26 i 0.67 17.36 i 0.24 130.92 < .01 212.00 i 0.50 15.49 i 0.33 129.08 <.01 10 3 8.81 i 0.79 29.20 i 0.54 331.44 <.01 410.82 i 0.63 12.98 i 0.58 119.96 < .01 18.38 i 0.34 169.87 5 7.78 i 0.22 9.25 i 0.48 118.89 < .05 617.32 i 0.93 22.35 ~ 0.39 129.04 < .01 7 6.37 i 0.89 18.69 ~ 1.27 294.98 < .01 15 8 9.63 i 0.77 14.55 i 1.17 151.09 <.05 914.01 i 0.89 20.09 i 0.20 143.40 < .01 10 6.06 i 0.83 18.51 i 0.89 305.22 <.01 Average10.61 i 3.6 17.91 i 5.2 184 HT-29SF 11.96 i 0.16 CA 0224l2~ l998-06-22 WO 97/23230 PCT/US96/~0649 T~BLE 3 Ex vivo patient data:
Preferential 5mAZC deamination to AZT in tumor vs. normal adjacent mucosa 5 Patient # Radiolabeled AZT Radiolabeled AZT Percentage p value in Normal in Colon TumorAZT in Tumor AdjacentTissue Tissue Tissue of pmol/min/mg pmol/min/mg Normal protein protein 117.14 i 7.61 134.51 i 14.92 114.83 After THU:
30.56 i 4.22 2 128.76 i 2.17 153.34 i 8.70 119.09 < .05 After THU:
50.48 i 7.30 3 40.42 i 1.18304.04 i 17.63 752.17 < .01 After THU:
0.31 i 0.31 4 77.52 i 8.8380.01 i 20.23 103.33 polyp After THU:
6.47 i 6.47 178.88 < .01 138.58 i 1.44 After THU:
34.83 + 2.17 144.28 i 20.39 225.21 ~ 44.91 156.09 After THU:
90.90 i 26.41 6 126.75 i 9.75 235.07 ~ 11.74 185.46 < .01 After THU:
74.84 i 6.51 7 70.58 i 16.31 164.62 + 20.63 233.23 < .05 After THU:
26.81 ~t 1.46 8 51.91 i 14.01 88.59 i 13.80 170.64 After THU:
61.31 i 4.57 9 89.05 i 13.05 132.10 i 4.83 148.34 < .05 After THU:
39.88 + 24.18 64.28 i 6.00 147.80 i 10.95 229.93 < .01 After THU:
80.34 ~ 16.86 Average 91.1 i36.0 172.5 i63 229
34.83 + 2.17 144.28 i 20.39 225.21 ~ 44.91 156.09 After THU:
90.90 i 26.41 6 126.75 i 9.75 235.07 ~ 11.74 185.46 < .01 After THU:
74.84 i 6.51 7 70.58 i 16.31 164.62 + 20.63 233.23 < .05 After THU:
26.81 ~t 1.46 8 51.91 i 14.01 88.59 i 13.80 170.64 After THU:
61.31 i 4.57 9 89.05 i 13.05 132.10 i 4.83 148.34 < .05 After THU:
39.88 + 24.18 64.28 i 6.00 147.80 i 10.95 229.93 < .01 After THU:
80.34 ~ 16.86 Average 91.1 i36.0 172.5 i63 229
Claims (50)
1. An agent having the chemical formula C7N3H8O2R1R2XX1, wherein X and X1 are each independently C or N but both may not be C; R1 is selected from the group consisting of lower alkyl, alkenyl and alkynyl, halogen and haloalkyl;
and R2 is selected from the group consisting of H, -N3, -OH, amino and halogen; or pharmaceutically acceptable salts thereof.
and R2 is selected from the group consisting of H, -N3, -OH, amino and halogen; or pharmaceutically acceptable salts thereof.
2. The agent of claim 1, wherein R1 is methyl and R2 is -N3.
3. The agent of claims 1 or 2, wherein R1 is CF3
4. The agent of claims 1 to 3, which is selected from the group consisting of 5-methyl-2', 3-dideoxycytidine, 5-ethyl-2',3' dideoxy-3' - azidocytidine, 5-propyl-2',3-dideoxycytidine, 5-propyl-2',3'-dideoxy-3-azidocytidine, 5-propene-2', 3'-dideoxy-3'-azidocytidine, 5-propyne-2',3'-dideoxy-3'-azidocytidine, 5-propyne-2',3'-dideoxycytidine and 5-propyne-2',3'-dideoxy-3'-azidocytidine.
5. The agent of claims 1 to 4, or pharmaceutically acceptable salts thereof, having the chemical formula
6. The agent of claims 1 to 5, wherein X comprises N and X1 comprises C.
7. The agent of claims 1 to 5, wherein X comprises C and X1 comprises N.
8. The agent of claims 1 to 5, wherein X comprises N and X1 comprises N.
9. The agent of claims 1 to 8, further comprising a radiolabel.
10. The agent of claims 1 to 9, which is freeze-dried or lyophilized.
11. The agent of claims 1 to 10, wherein the R1 haloalkyl is selected from the group consisting of alkyl substituted by one or more chloro, fluoro, bromo and iodo.
12. The agent of claims 1 to 11, wherein the R1 fluoroalkyl comprises -CF3.
13. A composition, comprising the agent of claims 1 to 12; and a pharmaceutically acceptable carrier.
14. The composition of claim 13, comprising about 0.5 to about 99 % of the agent.
15. The composition of claims 13 to 14, in unit dosage form.
16. The composition of claims 13 to 15, in multi-dosage form.
17. The composition of claims 13 to 16, in a form selected from the group consisting of capsules, cachets, pastilles, lozanges, powder, granules, solution, suspension, emulsion and tablets.
18. The composition of claims 13 to 17, wherein the carrier is selected from the group consisting of solid and liquid carriers.
19. The composition of claim 13 to 18, further comprising an agent selected from the group consisting of other therapeutic agents, flavorings, lubricants, suspending and thickening agents, binders, inert diluents, surface active agents, dispersants, antioxidants, buffers, bacteriostats and solutes to attain isotonicity.
20. The composition of claims 13 to 19, wherein the therapeutic agent comprises an anti-leukemia agent.
21. The composition of claims 13 to 20, wherein the anti-leukemia agent comprises cytosine arabinoside.
22. An oral formulation comprising the composition of claims 13 to 21, and an enteric coating.
23. A sub-lingual formulation comprising the composition of claims 13 to 22, wherein the flavoring and inert diluent are selected from the group consisting of sucrose, acacia, tragacanth, gelatin and glycerin.
24. A parenteral formulation comprising the composition of claims 13 to 21, comprising a solution, suspension or emulsion.
25. An ampule or vial comprising the formulation of claims 13 to 21 and 24.
26. A rectal formulation comprising the composition of claims 13 to 21, in unit dosage form.
27. A transdermal formulation comprising the composition of claims 13 to 21, and an iontophoretic medium.
28. A transdermal device, comprising a patch which comprises the formulation of claims 13 to 21 and 27.
29. An iontophoretic device comprising the transdermal device of claim 28, and means for iontophoretic delivery.
30. A .therapeutic kit, comprising in a sealed container the composition of claims 13 to 29, and instructions for its administration.
31. A method of preventing or treating a disorder associated with cytidine diaminase or deoxycytidine deaminase over-expression, comprising administering to a subject in need of such treatment an anti-disorder effective amount of an agent selected from the group consisting of the agent of the chemical formula C7N3H6O3R1R2XX1, wherein X and X1 are each independently C or N, R1 is selected from the group consisting of lower alkyl, alkenyl and alkynyl, halogen and haloalkyl, and R2 is selected from the group consisting of H, -N3,-OH, amino and halogen; or pharmaceutically acceptable salts thereof.
32. The method of claim 31, wherein R1 is methyl and R2 is -N3.
33. The method of claim 31 or 32, wherein the agent is selected from the group consisting of 5- methyl- 2', 3-dideoxycytidine, 5- ethyl- 2', 3' dideoxy- 3' -azidocytidine, 5- propyl- 2', 3- dideoxycytidine, 5-propyl-2',3'-dideoxy-3-azidocytidine, 5-propene-2', 3'-dideoxy-3'-azidocytidine, 5-propyne-2',3'-dideoxy-3'-azidocytidine, 5-propyne-2',3'-dideoxycytidine and 5-propyne-2',3'-dideoxy-3'-azidocytidine.
34. The method of claims 31 to 33, wherein the disorder is cytosine arabinoside (araC) -resistant leukemia.
35. The method of claims 31 to 34, further comprising administering to the subject an anti-leukemia effective amount of an anti-leukemia agent.
36. The method of claims 31 to 35, wherein the anti-leukemia agent is administered concurrently with the agent.
37. The method of claims 31 to 36, wherein the anti-leukemia agent comprises cytosine arabinoside (araC).
38. The method of claims 31 to 37 , wherein the disorder comprises arthritis.
39. The method of claims 31 to 38, which is a prophylactic method.
40. The method of claims 31 to 38, which is a therapeutic method.
41. The method of claims 31 to 40, wherein the agent is administered parenterally.
42. The method of claims 31 to 40 , wherein the agent is administered orally.
43. The method of claims 31 to 40, wherein the agent is administered transdermally.
44. The method of claims 31 to 43, wherein the agent is administered in an amount of about 0.1 to about 1000 µmol/kg body weight.
45. The method of claims 31 to 44, wherein the subject is human.
46. The method of claims 31 to 45, wherein the subject is a non-human animal.
47. The method of claims 31 to 46, wherein the non-human animal is selected from the group consisting of animals in the care of a veterinarian.
48. The method of claims 31 to 47, for treating a cancer associated with cytidine deaminase or deoxycytidine deaminase over-expression, wherein the agent is administered in an anti-cancer effective amount.
49. The method of claims 31 to 47, for preventing or treating an HIV infection, wherein the agent is comprising administered in an anti-HIV effective amount.
50. The method of claims 51 to 49, for reducing the number or toxicity of proinflammatory cells which over-express cytidine deaminase or deoxycytidine deaminase, wherein the agent is administered in an anti-inflammation effective amount.
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US57718595A | 1995-12-22 | 1995-12-22 | |
US08/577,185 | 1995-12-22 |
Publications (1)
Publication Number | Publication Date |
---|---|
CA2241255A1 true CA2241255A1 (en) | 1997-07-03 |
Family
ID=24307627
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CA002241255A Abandoned CA2241255A1 (en) | 1995-12-22 | 1996-12-23 | Method of treating disorders characterized by overexpression of cytidine deaminase or deoxycytidine deaminase |
Country Status (8)
Country | Link |
---|---|
US (1) | US6136791A (en) |
EP (1) | EP0876149A4 (en) |
JP (1) | JPH11513992A (en) |
KR (1) | KR19990076695A (en) |
AU (1) | AU728377C (en) |
CA (1) | CA2241255A1 (en) |
RU (1) | RU98113785A (en) |
WO (1) | WO1997023230A1 (en) |
Families Citing this family (19)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6432924B1 (en) * | 1993-12-26 | 2002-08-13 | East Carolina University | Method of treating disorders characterized by overexpression of cytidine deaminase or deoxycytidine deaminase |
US6982253B2 (en) * | 2002-06-05 | 2006-01-03 | Supergen, Inc. | Liquid formulation of decitabine and use of the same |
ES2538486T3 (en) * | 2002-09-12 | 2015-06-22 | International Institute Of Cancer Immunology, Inc. | Preparation of antigenic peptides against cancer |
EP1556010A4 (en) * | 2002-10-31 | 2007-12-05 | Supergen Inc | Pharmaceutical formulations targeting specific regions of the gastrointestinal tract |
US20060063735A1 (en) * | 2004-09-17 | 2006-03-23 | Supergen, Inc. | Salts of 5-azacytidine |
US20060069060A1 (en) * | 2004-09-27 | 2006-03-30 | Sanjeev Redkar | Salts of decitabine |
US20060128654A1 (en) * | 2004-12-10 | 2006-06-15 | Chunlin Tang | Pharmaceutical formulation of cytidine analogs and derivatives |
US20060128653A1 (en) * | 2004-12-10 | 2006-06-15 | Chunlin Tang | Pharmaceutical formulation of decitabine |
US7250416B2 (en) * | 2005-03-11 | 2007-07-31 | Supergen, Inc. | Azacytosine analogs and derivatives |
US7700567B2 (en) | 2005-09-29 | 2010-04-20 | Supergen, Inc. | Oligonucleotide analogues incorporating 5-aza-cytosine therein |
BR112014004779B1 (en) | 2011-08-30 | 2022-01-18 | Astex Pharmaceuticals, Inc | DECITABINE DERIVATIVES FORMULATIONS, KIT AND RELATED PROCESSES |
EP2820240A4 (en) | 2012-02-29 | 2015-10-14 | Koskan Larry P | System and method for inhibiting scale formation in oil wells |
CN105683209B (en) | 2013-10-29 | 2019-03-08 | 大塚制药株式会社 | 2 '-deoxidations -2 ', the route of synthesis of 2 '-difluoro tetrahydrouridines |
WO2016195353A1 (en) * | 2015-06-01 | 2016-12-08 | Kainos Medicine, Inc. | A use of 1'-cyano-cytarabine for cancer treatment |
WO2017004538A1 (en) | 2015-07-02 | 2017-01-05 | Otsuka Pharmaceutical Co., Ltd. | Lyophilized pharmaceutical compositions |
WO2017158396A1 (en) | 2016-03-16 | 2017-09-21 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Cytidine deaminase inhibitors for the treatment of pancreatic cancer |
US11209421B2 (en) * | 2016-03-31 | 2021-12-28 | Centre National De La Recherche Scientifique | Cytidine deaminase expression level in cancer as a new therapeutic target |
AU2018310857A1 (en) | 2017-08-03 | 2020-02-13 | Otsuka Pharmaceutical Co., Ltd. | Drug compound and purification methods thereof |
WO2023069529A1 (en) | 2021-10-19 | 2023-04-27 | Akirabio, Inc. | Compositions comprising 2'-deoxycytidine analogs and use thereof for the treatment of sickle cell disease, thalassemia, and cancers |
Family Cites Families (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE2122991C2 (en) * | 1971-05-04 | 1982-06-09 | Schering Ag, 1000 Berlin Und 4619 Bergkamen | Process for the preparation of cytosine and 6-azacytosine nucleosides |
US4501744A (en) * | 1980-09-16 | 1985-02-26 | Ens Bio Logicals Inc. | Substituted-aza-cytosine compounds and anti-viral uses thereof |
US5084445A (en) * | 1986-05-01 | 1992-01-28 | University Of Georgia Research Foundation, Inc. | 3'-azido-2',3'-dideoxy-5-methylcytidine |
US4788181A (en) * | 1986-09-29 | 1988-11-29 | The United States Of America As Represented By The Department Of Health And Human Services | 5-substituted-2',3'-dideoxycytidine compounds with anti-HTLV-III activity |
GB8629892D0 (en) * | 1986-12-15 | 1987-01-28 | Wellcome Found | Antiviral compounds |
US5215971A (en) * | 1986-12-19 | 1993-06-01 | Medivir Ab | Antiviral pharmaceutical composition comprising 5-substituted pyrimidine nucleosides |
SE8802173D0 (en) * | 1988-06-10 | 1988-06-10 | Astra Ab | PYRIMIDINE DERIVATIVES |
US5652099A (en) * | 1992-02-12 | 1997-07-29 | Conrad; Michael J. | Probes comprising fluorescent nucleosides and uses thereof |
-
1996
- 1996-12-23 RU RU98113785/14A patent/RU98113785A/en not_active Application Discontinuation
- 1996-12-23 KR KR1019980704809A patent/KR19990076695A/en not_active Application Discontinuation
- 1996-12-23 CA CA002241255A patent/CA2241255A1/en not_active Abandoned
- 1996-12-23 US US08/772,455 patent/US6136791A/en not_active Expired - Fee Related
- 1996-12-23 WO PCT/US1996/020649 patent/WO1997023230A1/en not_active Application Discontinuation
- 1996-12-23 EP EP96945043A patent/EP0876149A4/en not_active Withdrawn
- 1996-12-23 AU AU13501/97A patent/AU728377C/en not_active Ceased
- 1996-12-23 JP JP9523869A patent/JPH11513992A/en not_active Ceased
Also Published As
Publication number | Publication date |
---|---|
KR19990076695A (en) | 1999-10-15 |
EP0876149A1 (en) | 1998-11-11 |
WO1997023230A1 (en) | 1997-07-03 |
RU98113785A (en) | 2000-06-10 |
AU728377B2 (en) | 2001-01-11 |
US6136791A (en) | 2000-10-24 |
AU728377C (en) | 2003-10-30 |
EP0876149A4 (en) | 2001-09-26 |
AU1350197A (en) | 1997-07-17 |
JPH11513992A (en) | 1999-11-30 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CA2241255A1 (en) | Method of treating disorders characterized by overexpression of cytidine deaminase or deoxycytidine deaminase | |
JP5357857B2 (en) | Anticancer agent and DNA replication inhibitor | |
JP2003201240A (en) | Method for reducing toxicity of chemotherapeutic agent and antiviral agent with acylated pyrimidine nucleoside | |
AU667676B2 (en) | Treatment of chemotherapeutic agent and antiviral agent toxicity with acylated pyrimidine nucleosides | |
SK281231B6 (en) | Monoester compound, pharmaceutical composition it containing, and use of this compound | |
CA2574032A1 (en) | Compounds and methods for the treatment of cancer | |
WO1994026761A1 (en) | Treatment of chemotherapeutic agent and antiviral agent toxicity with acylated pyrimidine nucleosides | |
JP5730854B2 (en) | Combination of decitabine and cytidine deaminase inhibitor and its use in the treatment of cancer | |
US6432924B1 (en) | Method of treating disorders characterized by overexpression of cytidine deaminase or deoxycytidine deaminase | |
WO1996028170A1 (en) | Reduced toxicity compositions and methods for treating hiv infections | |
JPH10507441A (en) | Method for treating sepsis or inflammatory disease using oxypurine nucleosides | |
JPH06511473A (en) | Oxypurine nucleosides and their congeners and their acyl derivatives for improving hematopoiesis | |
JP2022525156A (en) | Modified microRNAs and their use in the treatment of cancer | |
JP2002515892A (en) | Treatment of cytokine-related diseases | |
AU732120B2 (en) | Pyrimidine nucleotide precursors for treatment of systemic inflammation and inflammatory hepatitis | |
JP2020517716A (en) | Blood cancer treatment | |
JPS62426A (en) | Therapy for neoplasmic disease | |
US7157449B2 (en) | Medicament for the treatment of diseases caused by parasitic protozoa | |
JP5581200B2 (en) | Antitumor agent containing cytidine derivative and carboplatin | |
WO2022014025A1 (en) | Novel therapeutic method and novel therapeutic agent for hematological cancer | |
TW202140018A (en) | Antitumor effect enhancing agent containing uracil derivative compound | |
JPH0413328B2 (en) | ||
CZ246999A3 (en) | Method of reducing administered dosage of a first medicament when treating disease |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
EEER | Examination request | ||
FZDE | Discontinued |