CA1199274A - TREATMENT OF MALIGNANT TUMORS WITH 2-.beta.-D- RIBOFURANOSYL-THIAZOLE-4-CARBOXAMIDE AND RELATED COMPOUNDS - Google Patents
TREATMENT OF MALIGNANT TUMORS WITH 2-.beta.-D- RIBOFURANOSYL-THIAZOLE-4-CARBOXAMIDE AND RELATED COMPOUNDSInfo
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Abstract
ABSTRACT
The compound 2-.beta.-D-ribofuranosylthiazole-4-carboxamide is used to treat malignant tumors in warm blooded animals.
Esters of this compound such as 2-(2,3,5-tri-O-acetyl-.beta.-D-ribofuranosyl) thiazole-4-carboxamide and 2-(5-O-phosphoryl-.beta.-D-ribofuranosyl)-thiazole-4-carboxamide are also useful for treating tumors in warm blooded animals.
The compound 2-.beta.-D-ribofuranosylthiazole-4-carboxamide is used to treat malignant tumors in warm blooded animals.
Esters of this compound such as 2-(2,3,5-tri-O-acetyl-.beta.-D-ribofuranosyl) thiazole-4-carboxamide and 2-(5-O-phosphoryl-.beta.-D-ribofuranosyl)-thiazole-4-carboxamide are also useful for treating tumors in warm blooded animals.
Description
3~ 1 This is' a divisional application of copendin~ app1icatio r-~Seri~l No. 392,323, filed Decer~er 15, 198].
BAC:~GROUND OF THE INVENTION
This invention is directed -to a -treatment oE
malignant -tumors in vivo using the compound 2-~-D-ribo-fuxanosylthiazole-4-carboxamide and related deriva-~ives such as its estexs.
Control of malignant tumors in rnan and animals still remains as an unrealized goal. Within the last several decades, understanding of malignancy has made significant progress;
however, conquering of the malignant disease state has not been realized.
Con~entional therapy of both humans and other valuable animal species inflicted with malignant tumors presently includes surgical excising of the tumor, local radiation therapy oiF the afflicted animal, and chemotherapy by administration oF a chemctheraputic agent to the animal. The death of a significant number of patien-ts inflicted with malignant tumors is attributable no-t to the primary tumor bu-t instead to metastasis o~ the primary tumor -to secondary sites in the host.
If a primary tumor is detected early, it normally can be eliminated by surgery, radiation or chemotherapy or combinations of these. The metastatic colonies of these primary tumors, however, are exceedingly harder to detect and eliminate and the unsuccessful managemen-t of them remains a serious medical problem.
Tumors are normally classified either as benign or malignant. The mali~nant tumor is characterized -From the benign by its ability to invade both surrounding -tissue and to coloni2e distant sites via metas-tasis~ Certain organs are more prone -to metastasis -than o-thers. Included in this qroup would be the lung, the brain, the liver, the ovaries and -the adrenal glands. It has further been suggested that both surgery and radiation of a primary tumor in certain instances actually promotes metastasis.
In view of the inability of current cancer therapy to successfully control the malignant tumor and its metastasis, it is evident that there exists a need for additional chemotheraputic agents.
In a paper entitled Synthesis and Antiviral Activity of Certain Thiazole C-Nucleosides, J. Med. Chem. 1977, Volume 20, No. 2, 256, I and my co-workers disclosed the synthesis of and certain preliminary in vitro antiviral activity of the compounds 2-.beta.-D-ribofuranosylthiazole-4-carboxamide and 2-(2,3,5-tri-O-acetyle-.beta.-D-ribofuranosyl)-thiazole-4-carboxamide in an in vitro test system utilizing three viruses, type 1 herpes simplex virus, type 3 para-influenza virus and type 3 rhinovirus. The in vitro activity of the compound 2-.beta.-D-ribofuranosylthiazole-4-carboxamide against these three viruses was only moderate. With the compound 2-(2,3,5-tri-O-acetyle-.beta.-D-ribofuranosyl)thiazole-4-carboxamide, only moderate activity was seen with type 1 herpes simplex virus whereas with the type 3 parainfluenza and the type 3 rhinoviruses activity was negative. While certain marginal in vitro antiviral activity noted in the preceding was seen, quite to the contrary, in vivo antiviral testing for both 2-.beta.-D-ribofuranosylthiazole-4-carboxamide and 2-(2,3,5-tri-O-acetyl-.beta.-D-ribofuranosyl)thiazole-4-carboxamide, as judged by the number of test animal deaths, was negative. In the in vivo tests, the number of deaths for the test animals for both 2-.beta.-D-ribofuranosylthiazole-4-carboxamide and 2-(2,3,5-tri-O-acetyl-.beta.-D-ribofuranosyl)-
BAC:~GROUND OF THE INVENTION
This invention is directed -to a -treatment oE
malignant -tumors in vivo using the compound 2-~-D-ribo-fuxanosylthiazole-4-carboxamide and related deriva-~ives such as its estexs.
Control of malignant tumors in rnan and animals still remains as an unrealized goal. Within the last several decades, understanding of malignancy has made significant progress;
however, conquering of the malignant disease state has not been realized.
Con~entional therapy of both humans and other valuable animal species inflicted with malignant tumors presently includes surgical excising of the tumor, local radiation therapy oiF the afflicted animal, and chemotherapy by administration oF a chemctheraputic agent to the animal. The death of a significant number of patien-ts inflicted with malignant tumors is attributable no-t to the primary tumor bu-t instead to metastasis o~ the primary tumor -to secondary sites in the host.
If a primary tumor is detected early, it normally can be eliminated by surgery, radiation or chemotherapy or combinations of these. The metastatic colonies of these primary tumors, however, are exceedingly harder to detect and eliminate and the unsuccessful managemen-t of them remains a serious medical problem.
Tumors are normally classified either as benign or malignant. The mali~nant tumor is characterized -From the benign by its ability to invade both surrounding -tissue and to coloni2e distant sites via metas-tasis~ Certain organs are more prone -to metastasis -than o-thers. Included in this qroup would be the lung, the brain, the liver, the ovaries and -the adrenal glands. It has further been suggested that both surgery and radiation of a primary tumor in certain instances actually promotes metastasis.
In view of the inability of current cancer therapy to successfully control the malignant tumor and its metastasis, it is evident that there exists a need for additional chemotheraputic agents.
In a paper entitled Synthesis and Antiviral Activity of Certain Thiazole C-Nucleosides, J. Med. Chem. 1977, Volume 20, No. 2, 256, I and my co-workers disclosed the synthesis of and certain preliminary in vitro antiviral activity of the compounds 2-.beta.-D-ribofuranosylthiazole-4-carboxamide and 2-(2,3,5-tri-O-acetyle-.beta.-D-ribofuranosyl)-thiazole-4-carboxamide in an in vitro test system utilizing three viruses, type 1 herpes simplex virus, type 3 para-influenza virus and type 3 rhinovirus. The in vitro activity of the compound 2-.beta.-D-ribofuranosylthiazole-4-carboxamide against these three viruses was only moderate. With the compound 2-(2,3,5-tri-O-acetyle-.beta.-D-ribofuranosyl)thiazole-4-carboxamide, only moderate activity was seen with type 1 herpes simplex virus whereas with the type 3 parainfluenza and the type 3 rhinoviruses activity was negative. While certain marginal in vitro antiviral activity noted in the preceding was seen, quite to the contrary, in vivo antiviral testing for both 2-.beta.-D-ribofuranosylthiazole-4-carboxamide and 2-(2,3,5-tri-O-acetyl-.beta.-D-ribofuranosyl)thiazole-4-carboxamide, as judged by the number of test animal deaths, was negative. In the in vivo tests, the number of deaths for the test animals for both 2-.beta.-D-ribofuranosylthiazole-4-carboxamide and 2-(2,3,5-tri-O-acetyl-.beta.-D-ribofuranosyl)-
- 2 --thiazole~4-carboxamide was equal to or exceeded the number of deaths of -the p:Lacebo control arlimals indicatincJ tha-t both of the compounds 2-~ D--ribofuranosylthiazole-4-carboxamide and 2-(2~3,5-tri-O-ace-tyl- ~-D-ribofuranosyl~thiazole-4-carboxamide demonstrated no useful in vivo antiviral activity.
With regard to the above noted in vitro antiviral testing of both 2- ~-D-ribofuranosylthiazole-4-carboxamide and 2-(2,3,5-tri-O-acetyl ~-D~ribvfuranosyl)thiazole-4-carboxamide, these compounds were te~ted against viruses or which the known antiviral compound RIBAVIRIN~ i5 known to have positive antivir~. activity. In view of ~he preliminary marginal in vitro activity of 2- ~-D-ribofuranosyl-thiazole-4 carboxamide against these test viruses, it was expected that the spectrum of activity of 2- ~-D-ribofuranosylthiazole-4-carboxamide would be similar to the spectrum of activity of the compound RIBAVIRIN~. RIBAVIRIN is kno~n to be an active in vi-tro an-tiviral agent and in vivo antiviral and is further known to exhibit no siynificant antitumor ac-tivity~ Additionally, cer-tain derivatives of RIBAVIRIN~ such as its 5' monophosphate are also known to be inactive as antitumor compounds~ I-t was reasonable to expect, in comparing the preliminary în vitro antiviral activity of 2-D-ribofurallosylthiazole-4-carboxamide with that of RIBAVIRIN~, that ~-~-D-ribofurano~ylthiazole-4-carboxamide would exhibit positive in vivo antiviral activity and negative ~ntitumor activity similar to RIBA~IRIN~. Totally, contrary to this, the compound 2 ~-D-ribofuranosylthia~ole-4 carboxamide possessed no useful in vivo antivi.ral. ~ctivity and, quit.e unexpectedly, has demonstrated positive antitumor ac-tivity.
_ ,~ , . ..
i~q~''3~ ~
I have found that t:he compound 2-~-D-ribofurano-sylthiazole-4-carboxamide and its es-ters, including 2-(2,3,5--tri-O-acetyl~ D-riboEuranosyl)thiazole-4-carbo~a~ide and 2-(5~0 phosphoryl~-D-ribofuranosy:l)thiazole~-c~rboxamide exhibi-t antitumor activities of such significance as to be useful as antitumor agents in vivo.
BRIEF SUMl~RY OF THE INVENTION
The compound 2-~-D-ribofuranosyltniazole~4-carboxamide has been shown to exnibit significan-t antit7~mor activity in vivoO The present invention relates to the use of this compolmd and certain related derivatives in treating maligna~t tumors in warm blooded animals. According to this invention, the antitumor properties of 2-~-D-ribofuranosylthiazole~4 caxboxamide and its related esters are utilized ~y administerin~
to a warm blooded animal an effective amount of a pharmaceutical composition containing as the active compound at least about 0~1 percent by weight, based on the total weight of the composition, a compound of the structure:
2 0 2N >~\
N~,S
R30 _ / O \
\~ ~
R ?
wherein Rl and R2 are H or Cl-C18 aoyl and R3 is Hl Cl~C18 acyl or HQ ?- and physioloaically acceptable sal~s -thereo.
7~
In .l more preferred ~roup of compounds Rl and R2 are ~1 or ,~
Cl C8 acyl and R3 is H, Cl~C8 acyl or ~ and physiologically OH
aceeptable salts thereof.
Specifically noted for Rl, R2 and R3 as preferred acyl groups are acetyl, propionyl, isobu-tyryl and b~nzoyl.
Specifically noted as acceptable salts are the alkalai metals and ammonium or substituted ammonium salts sueh as sodium, pota~sium and ammonium ~alts.
Preferrably, when Rl and R2 are H, R3 is OH, Cl-C8 aeyl or HO-?~ , and when Rl and R2 is Cl-C~ aeyl, R3 is Cl-C8 aeyl~
For use in the pharmaceutieal composition of the invention, a pharmaceutieal carrier would be utili~ed sueh -that, preferredly, the pharmaceutical carrier would be ehosen -to allow administration of a suitable concentration of the ac-tive compounds of the invention as solutions or suspensions by injec~ion into an afflicted warm blooded animal. Depending on the host harboring the malignant tumor, the type of tumor, and the tumor site, administration by injeetion would be intraveneously, intra~uscularly, intracerebrally~ subeutaneously, or intraperitoneally.
Alternately r the eomposition of the invention mi~ht suitably be formulated in appropriate pharmaeeutical earriers allowing for adminis-tration by other routes such as oral administration, ophthalmic administration, topical administration or aclministration by suppository~
DETAILED DESCRlPTION
The parerlt compound of the invention, compound 2-~-U~
pC/ l''~' '' ~ ~g~ g ~
rlboEuranosylthiazo:le~4-carboxamicle, is preferredly prepared as described in Example l. An al-ternate sy~-thesis oE this compound appears in J. Org. Chem. r Vol~ 41~ ~0. 26 1976, 4074~
Cer-tain es-ters o:E 2-~-D-ribofuranosylthiazQ,le~4-carboxamide, compound 1, such as 2-(2,3,5-tri-O-acet~l-~-D-ri.bofuranosyl)thiazol.e-4-carboxamide, compound 2~ or 2-~5 O~
phosphoryl-~-D-ribofuraIlosyl~thiazole-4-carboxamide r compound
With regard to the above noted in vitro antiviral testing of both 2- ~-D-ribofuranosylthiazole-4-carboxamide and 2-(2,3,5-tri-O-acetyl ~-D~ribvfuranosyl)thiazole-4-carboxamide, these compounds were te~ted against viruses or which the known antiviral compound RIBAVIRIN~ i5 known to have positive antivir~. activity. In view of ~he preliminary marginal in vitro activity of 2- ~-D-ribofuranosyl-thiazole-4 carboxamide against these test viruses, it was expected that the spectrum of activity of 2- ~-D-ribofuranosylthiazole-4-carboxamide would be similar to the spectrum of activity of the compound RIBAVIRIN~. RIBAVIRIN is kno~n to be an active in vi-tro an-tiviral agent and in vivo antiviral and is further known to exhibit no siynificant antitumor ac-tivity~ Additionally, cer-tain derivatives of RIBAVIRIN~ such as its 5' monophosphate are also known to be inactive as antitumor compounds~ I-t was reasonable to expect, in comparing the preliminary în vitro antiviral activity of 2-D-ribofurallosylthiazole-4-carboxamide with that of RIBAVIRIN~, that ~-~-D-ribofurano~ylthiazole-4-carboxamide would exhibit positive in vivo antiviral activity and negative ~ntitumor activity similar to RIBA~IRIN~. Totally, contrary to this, the compound 2 ~-D-ribofuranosylthia~ole-4 carboxamide possessed no useful in vivo antivi.ral. ~ctivity and, quit.e unexpectedly, has demonstrated positive antitumor ac-tivity.
_ ,~ , . ..
i~q~''3~ ~
I have found that t:he compound 2-~-D-ribofurano-sylthiazole-4-carboxamide and its es-ters, including 2-(2,3,5--tri-O-acetyl~ D-riboEuranosyl)thiazole-4-carbo~a~ide and 2-(5~0 phosphoryl~-D-ribofuranosy:l)thiazole~-c~rboxamide exhibi-t antitumor activities of such significance as to be useful as antitumor agents in vivo.
BRIEF SUMl~RY OF THE INVENTION
The compound 2-~-D-ribofuranosyltniazole~4-carboxamide has been shown to exnibit significan-t antit7~mor activity in vivoO The present invention relates to the use of this compolmd and certain related derivatives in treating maligna~t tumors in warm blooded animals. According to this invention, the antitumor properties of 2-~-D-ribofuranosylthiazole~4 caxboxamide and its related esters are utilized ~y administerin~
to a warm blooded animal an effective amount of a pharmaceutical composition containing as the active compound at least about 0~1 percent by weight, based on the total weight of the composition, a compound of the structure:
2 0 2N >~\
N~,S
R30 _ / O \
\~ ~
R ?
wherein Rl and R2 are H or Cl-C18 aoyl and R3 is Hl Cl~C18 acyl or HQ ?- and physioloaically acceptable sal~s -thereo.
7~
In .l more preferred ~roup of compounds Rl and R2 are ~1 or ,~
Cl C8 acyl and R3 is H, Cl~C8 acyl or ~ and physiologically OH
aceeptable salts thereof.
Specifically noted for Rl, R2 and R3 as preferred acyl groups are acetyl, propionyl, isobu-tyryl and b~nzoyl.
Specifically noted as acceptable salts are the alkalai metals and ammonium or substituted ammonium salts sueh as sodium, pota~sium and ammonium ~alts.
Preferrably, when Rl and R2 are H, R3 is OH, Cl-C8 aeyl or HO-?~ , and when Rl and R2 is Cl-C~ aeyl, R3 is Cl-C8 aeyl~
For use in the pharmaceutieal composition of the invention, a pharmaceutieal carrier would be utili~ed sueh -that, preferredly, the pharmaceutical carrier would be ehosen -to allow administration of a suitable concentration of the ac-tive compounds of the invention as solutions or suspensions by injec~ion into an afflicted warm blooded animal. Depending on the host harboring the malignant tumor, the type of tumor, and the tumor site, administration by injeetion would be intraveneously, intra~uscularly, intracerebrally~ subeutaneously, or intraperitoneally.
Alternately r the eomposition of the invention mi~ht suitably be formulated in appropriate pharmaeeutical earriers allowing for adminis-tration by other routes such as oral administration, ophthalmic administration, topical administration or aclministration by suppository~
DETAILED DESCRlPTION
The parerlt compound of the invention, compound 2-~-U~
pC/ l''~' '' ~ ~g~ g ~
rlboEuranosylthiazo:le~4-carboxamicle, is preferredly prepared as described in Example l. An al-ternate sy~-thesis oE this compound appears in J. Org. Chem. r Vol~ 41~ ~0. 26 1976, 4074~
Cer-tain es-ters o:E 2-~-D-ribofuranosylthiazQ,le~4-carboxamide, compound 1, such as 2-(2,3,5-tri-O-acet~l-~-D-ri.bofuranosyl)thiazol.e-4-carboxamide, compound 2~ or 2-~5 O~
phosphoryl-~-D-ribofuraIlosyl~thiazole-4-carboxamide r compound
3, are prepared as described in Examples 2 an~ 3 res~ctively.
1~ Additional.ly, other esters, such as the monoester 2~ O-acetyl-~-D-ribofuranosyl)thiazole-4-carboxamide, comp~und 4~
would be prepared as in the synthesis described in e~mple 4.
For other preferred carboxylic esters of the inventio~
substitution of acetic anhydrîde with a sui-ta~le an~ride such as propionic anhydride, butyric anhydri~e or ~e~oic anhydride is made. Alternately, the appropriate aci~ chloride could be substituted for the acid anhydride.
The esters of compound 1 could assist in de~ivery of the compound in ar. afflicted host. Such esters of the compound could be formed by reacting olle or more of t~e hydroxyl groups of the sugar moiety of compound 1 wit~ suitable reversible blocking groups which could be cleaved in ~ivo from the parent compound 1 by certain in situ chemical or ~nzymatic reactions.
For reaction with the hydroxyl groups/ es-te~s such as~ but not necessarily limited to, acyl and phosphor~l esters would be cons.idered The acyl groups can be ~elected from a group consisting of straight chain, branch chain, su~-tituted~
unsaturat.ed, saturated or aromatic acids such as, but not ~~ necessarily limited to, acetic, trifluoroacetic, propiLollic, (~
n-butyric, isob-ltyric, valeric, caproict pelargonic, enan-thic capyxilic, lactic, acrylic, propargylic, palmitic~ benzoic, phthalic, sa:Licylic~ cinnamlc and naphthoic acids. If a phosphoryl group is chosen, the phosphoryl ester could be as a free acid or as a salt form. Acceptable salts of the phcsphate moiety of the phosphoryl ester can be selected from, but not necessarily limited to, the group consisting of alkali and alkaline earths, e.g., sodium, potassium~
calcium, magnesium, lithium~ ammonium and subs-tituted ammonium, -trialkylammonium~ dialky'lammonium, alkylammonium~
e.g., triethylammonium, trimethylammonium, diethylammonium~
octylammonium, cetyltrimethylammonium and cetylpyridium.
As preferred forms of the esters of the inventi~n, compounds 2, 3 and 4 are mentioned. In addi-tion to these, other tri-O-acyl esters such as the 2',3',5'~tri-O'ben~oyl are mentioned. Additionally, other mono-es-ters such as the 5'-O-benzoyl is mentioned. Generally, for carboxylic esters the preferred esters would include Cl C18 acyls. A more preferred group includes Cl-C8 acylsO Preferredly, when phosphoryl esters ~ere utilized, the phosphate groups would be formed as a salt preferredly as a sodium salt or other alkali metal s~lt or ammonium.
Ester forms of compound 1, as is shown in the examples herein, are useful in delivering the compoun~ to the af~ected site in an affected host. As is shown in the examplesj the tri-acetyl ester, compounc~ 2~ is indicated as being an efEective antitumor agent when injected intra-peri~oneally into an affected host. The described triacetyl compound and any other acyl ester of compound 1 woulc~ ~e c~p^.ctecl -to be hydrol.i7.ed to compollncl ] in certain bioloyicaL
fluids such as the acid environrne.lt of the s-tomach or an environment which inclwcles an appropriate enZyMe capable o:~
in vivo enzymatic cleaY~cJ2 of ~he ester -to compound 1. While I do not wish -to be bound by theory, if the phosphoryl ester of compound I~ such as the 5' phosphate~ were used~ other en2ymes present in vivo migh-t also be expected to appropriately en~ymatically cleave the phosphate -to yield an in situ delivery of compound lo Compound 3, the phosphoryl ester of compound 1, as is shown in the examples~ is indicated as being an effective antitumor a~ent when injected in an effected hostO At this time it is not known whe-ther its ac-tivity is expressed as the 5I phcsphate or whether it i~ enzymatically cleaved to compound ]. Further, it is possible khat compound 1 might be promoted in situ by other enzymatic reactions to compound 3. In any event, both compound 1 and compound 3 are indicated as being effective in vivo antitumor ayents as is indicated by the examples.
In performing the invention, compound 1, or a selected ester form thereof, is appropriately mixed with a suitable pharmaceutical carrier which may be as si~ple as sterilized water or could be a complex carrier havin~ appropriate agents to suita~ly mimic certain biological environments J
i.e~, pH and salt adjusted solution sui-table for i~traveneou~, intramuscular or other injections.
In selecting a suitable pharmaceutiGal carrier, consideration of the type of tumor/ the site of the tumor and the health and age of the host would be gi~en. Ad~itionally, if an ester :Eorm o compound 1 is used~ considerati~n of the pc/~
chemical reactivity o:E the est-er would also he t3iven Thus, carbo~ylic acyl ester wou:Ld preferredly be suspended or solubilized in an appropriate non-clcidic medium. A phosphoryl ester miyht be appropriately used in the presence of a suitable buffer or as a salt as discussed above.
Preferredly, compound 1 or any of the other ~ompounds of the invention, would be mixed with an appropriate pharmaceutical carrier such that compound l or a derivative thereof would be suitably soluble in the carrier. Alternately, however, suspensions, emulsions and other formulations of the compounds of the invention could be used where indicated.
The pharmaceutical carrier, in addition to having a ~lubilizing or suspending agent therein, might also include ~ui~a~le dilutants, buffers, surface active agents and other similar agents as are typically used in pharmaceu-tical carriers. The total composition of the pharmaceutical carrier would, however, be chosen to be compatible with the site of delivery, 1the concentration of -the active ingredient and other parameters as are s'andard in pharmaceutical industry.
Compound l, or the other compounds of the invention, would be suitably admixed with the pharmaceutical carrier such that i~ would be present in a concentrat.ion of at least 0.1 percent by weight of the total composition. Preferredly, it would be present in the pharmaceutical carrier at a concentration oE about 10% to about 90% by we-ght o~ the total composition~
Effective amounts oE compound l, or the other compounds of the invention, typically would range f.rom about 2~5 milligrams per kilogram (mg/Kg) of the total body weight P~ P
~ 7 ~
oE the -treated warm blooded animal -to about 200 mg/K~ per day.
Preferredly, the ran~e would be from 12.5 mg/Kg to about 100 mg/Kg An even more preferred range would be from about 15 mg/K$ to about 50 mg~Kg. As with other fackors noted above, the amount of compound utilized in treating an afflicted animal would take into account parameters such as the type of tumor~ the t~mor site 9 the form of administerin~ the compound and the physical size and condition of the host~ In any event, the actual amount should be sufficient to provide a chemothera-peut.ically effective amount of the agent to the host in a convenient vol~ne, which will be readily within the ability of thos~ skilled in the art to determine given the disclosure herein.
In at least one study, compound 1 of the in~ention has been injected at dosages up to 2000 mg/Kg in-to tumor bearing animals and no deaths of the animals were attributed to the toxicity of compound 1 on the toxicity day of the test.
In a host which has been diagnosed as being terminally ill with a malignant tumor excessive amounts beyond any toxicity range might be indica-ted if there is ~ny probability of cure of the terminally ill host as is commonly practiced in current cancer chemotherapy.
As in the examples used for illustra-tive purposes below, wherein a tumor bearing host was treated once daily with the indicated test compound. Depending upon the clinical situation, the daily dose of compound 1 or any o~ the other ompounds of the invention~ might be similarly given; however, the daily dose cou:ld also be broken up into sub-unit doses mab/ ,l~
which, in their totality, e~ual ~he daily dose~ Thus r for example, at a 50 mg/Kg dose level the patient might be appropriately treated follr times a day with doses of 12~5 rng/KgO
A composition used Eor inhibiting ma~ignant tumors in warm blooded animals might be suitably pr pared by adding compound 1 or any of the other compounds o the inven-tion~ to a pharmacologically compatible solvent followed by sterilization and packaging in appropriate sealable vials at a known concentration. Appropriate doses of the compound are then withdrawn from the vial and administered by injection to -the host.
2-~-D-Ribofuranosylthia~ole~4--Carboxamide, COMPOUND 1 Ethyl 2-(2,3,5-tri V-benzoyl-~-D-ribofuranosyl)-thiazole-4-carboxamide was utilized as prepared in Srivastova et. al. J. Medu Chem. 1977, Volume 20, No. 2, 256.
concentrated solution of ethyl 2-(2/3~5-tri-o-benzyl-B-D-rib furanosyl)thia~ole-4-carboxamide (5O0 g, 8.31 mmol) in methanol (15 ml) was stirred with methanolic ammonia (satur-ated at 0C, 100 ml) in a pressure bottle at room temperature for 2 days. The solvent was evaporated and the residue wa~
chromatographed through a column (2.5 x 35 cm3 o silica gel (100 g) packed in ethyl acetate. E-~ution of the column with a solvent system (eth~l acetate-l propanol-water, 4:1:2; v~v;
top layer3 removed tne fast-moving methyl benzo~te and benz-amide~ The slower moving, major, W and sugar-positive fractions were collected and the solvent was evaporated in vacuo. The residue (syrup), thus obtained, was readily crys-tallized from e-th~nol-ethyl ace-tate to provide 106 g (74%~
of pure product, compound 1: mp 144-145 C.; la~ Sp-1'I.3 ( 1 DMF); W ~ pHl 237 nm (8640); UV ~maXP
H ~R(Me2SOd~)~7.5 7.8 [S(br),2,CONH2]~ Me2SO-d6-D2O~-
1~ Additional.ly, other esters, such as the monoester 2~ O-acetyl-~-D-ribofuranosyl)thiazole-4-carboxamide, comp~und 4~
would be prepared as in the synthesis described in e~mple 4.
For other preferred carboxylic esters of the inventio~
substitution of acetic anhydrîde with a sui-ta~le an~ride such as propionic anhydride, butyric anhydri~e or ~e~oic anhydride is made. Alternately, the appropriate aci~ chloride could be substituted for the acid anhydride.
The esters of compound 1 could assist in de~ivery of the compound in ar. afflicted host. Such esters of the compound could be formed by reacting olle or more of t~e hydroxyl groups of the sugar moiety of compound 1 wit~ suitable reversible blocking groups which could be cleaved in ~ivo from the parent compound 1 by certain in situ chemical or ~nzymatic reactions.
For reaction with the hydroxyl groups/ es-te~s such as~ but not necessarily limited to, acyl and phosphor~l esters would be cons.idered The acyl groups can be ~elected from a group consisting of straight chain, branch chain, su~-tituted~
unsaturat.ed, saturated or aromatic acids such as, but not ~~ necessarily limited to, acetic, trifluoroacetic, propiLollic, (~
n-butyric, isob-ltyric, valeric, caproict pelargonic, enan-thic capyxilic, lactic, acrylic, propargylic, palmitic~ benzoic, phthalic, sa:Licylic~ cinnamlc and naphthoic acids. If a phosphoryl group is chosen, the phosphoryl ester could be as a free acid or as a salt form. Acceptable salts of the phcsphate moiety of the phosphoryl ester can be selected from, but not necessarily limited to, the group consisting of alkali and alkaline earths, e.g., sodium, potassium~
calcium, magnesium, lithium~ ammonium and subs-tituted ammonium, -trialkylammonium~ dialky'lammonium, alkylammonium~
e.g., triethylammonium, trimethylammonium, diethylammonium~
octylammonium, cetyltrimethylammonium and cetylpyridium.
As preferred forms of the esters of the inventi~n, compounds 2, 3 and 4 are mentioned. In addi-tion to these, other tri-O-acyl esters such as the 2',3',5'~tri-O'ben~oyl are mentioned. Additionally, other mono-es-ters such as the 5'-O-benzoyl is mentioned. Generally, for carboxylic esters the preferred esters would include Cl C18 acyls. A more preferred group includes Cl-C8 acylsO Preferredly, when phosphoryl esters ~ere utilized, the phosphate groups would be formed as a salt preferredly as a sodium salt or other alkali metal s~lt or ammonium.
Ester forms of compound 1, as is shown in the examples herein, are useful in delivering the compoun~ to the af~ected site in an affected host. As is shown in the examplesj the tri-acetyl ester, compounc~ 2~ is indicated as being an efEective antitumor agent when injected intra-peri~oneally into an affected host. The described triacetyl compound and any other acyl ester of compound 1 woulc~ ~e c~p^.ctecl -to be hydrol.i7.ed to compollncl ] in certain bioloyicaL
fluids such as the acid environrne.lt of the s-tomach or an environment which inclwcles an appropriate enZyMe capable o:~
in vivo enzymatic cleaY~cJ2 of ~he ester -to compound 1. While I do not wish -to be bound by theory, if the phosphoryl ester of compound I~ such as the 5' phosphate~ were used~ other en2ymes present in vivo migh-t also be expected to appropriately en~ymatically cleave the phosphate -to yield an in situ delivery of compound lo Compound 3, the phosphoryl ester of compound 1, as is shown in the examples~ is indicated as being an effective antitumor a~ent when injected in an effected hostO At this time it is not known whe-ther its ac-tivity is expressed as the 5I phcsphate or whether it i~ enzymatically cleaved to compound ]. Further, it is possible khat compound 1 might be promoted in situ by other enzymatic reactions to compound 3. In any event, both compound 1 and compound 3 are indicated as being effective in vivo antitumor ayents as is indicated by the examples.
In performing the invention, compound 1, or a selected ester form thereof, is appropriately mixed with a suitable pharmaceutical carrier which may be as si~ple as sterilized water or could be a complex carrier havin~ appropriate agents to suita~ly mimic certain biological environments J
i.e~, pH and salt adjusted solution sui-table for i~traveneou~, intramuscular or other injections.
In selecting a suitable pharmaceutiGal carrier, consideration of the type of tumor/ the site of the tumor and the health and age of the host would be gi~en. Ad~itionally, if an ester :Eorm o compound 1 is used~ considerati~n of the pc/~
chemical reactivity o:E the est-er would also he t3iven Thus, carbo~ylic acyl ester wou:Ld preferredly be suspended or solubilized in an appropriate non-clcidic medium. A phosphoryl ester miyht be appropriately used in the presence of a suitable buffer or as a salt as discussed above.
Preferredly, compound 1 or any of the other ~ompounds of the invention, would be mixed with an appropriate pharmaceutical carrier such that compound l or a derivative thereof would be suitably soluble in the carrier. Alternately, however, suspensions, emulsions and other formulations of the compounds of the invention could be used where indicated.
The pharmaceutical carrier, in addition to having a ~lubilizing or suspending agent therein, might also include ~ui~a~le dilutants, buffers, surface active agents and other similar agents as are typically used in pharmaceu-tical carriers. The total composition of the pharmaceutical carrier would, however, be chosen to be compatible with the site of delivery, 1the concentration of -the active ingredient and other parameters as are s'andard in pharmaceutical industry.
Compound l, or the other compounds of the invention, would be suitably admixed with the pharmaceutical carrier such that i~ would be present in a concentrat.ion of at least 0.1 percent by weight of the total composition. Preferredly, it would be present in the pharmaceutical carrier at a concentration oE about 10% to about 90% by we-ght o~ the total composition~
Effective amounts oE compound l, or the other compounds of the invention, typically would range f.rom about 2~5 milligrams per kilogram (mg/Kg) of the total body weight P~ P
~ 7 ~
oE the -treated warm blooded animal -to about 200 mg/K~ per day.
Preferredly, the ran~e would be from 12.5 mg/Kg to about 100 mg/Kg An even more preferred range would be from about 15 mg/K$ to about 50 mg~Kg. As with other fackors noted above, the amount of compound utilized in treating an afflicted animal would take into account parameters such as the type of tumor~ the t~mor site 9 the form of administerin~ the compound and the physical size and condition of the host~ In any event, the actual amount should be sufficient to provide a chemothera-peut.ically effective amount of the agent to the host in a convenient vol~ne, which will be readily within the ability of thos~ skilled in the art to determine given the disclosure herein.
In at least one study, compound 1 of the in~ention has been injected at dosages up to 2000 mg/Kg in-to tumor bearing animals and no deaths of the animals were attributed to the toxicity of compound 1 on the toxicity day of the test.
In a host which has been diagnosed as being terminally ill with a malignant tumor excessive amounts beyond any toxicity range might be indica-ted if there is ~ny probability of cure of the terminally ill host as is commonly practiced in current cancer chemotherapy.
As in the examples used for illustra-tive purposes below, wherein a tumor bearing host was treated once daily with the indicated test compound. Depending upon the clinical situation, the daily dose of compound 1 or any o~ the other ompounds of the invention~ might be similarly given; however, the daily dose cou:ld also be broken up into sub-unit doses mab/ ,l~
which, in their totality, e~ual ~he daily dose~ Thus r for example, at a 50 mg/Kg dose level the patient might be appropriately treated follr times a day with doses of 12~5 rng/KgO
A composition used Eor inhibiting ma~ignant tumors in warm blooded animals might be suitably pr pared by adding compound 1 or any of the other compounds o the inven-tion~ to a pharmacologically compatible solvent followed by sterilization and packaging in appropriate sealable vials at a known concentration. Appropriate doses of the compound are then withdrawn from the vial and administered by injection to -the host.
2-~-D-Ribofuranosylthia~ole~4--Carboxamide, COMPOUND 1 Ethyl 2-(2,3,5-tri V-benzoyl-~-D-ribofuranosyl)-thiazole-4-carboxamide was utilized as prepared in Srivastova et. al. J. Medu Chem. 1977, Volume 20, No. 2, 256.
concentrated solution of ethyl 2-(2/3~5-tri-o-benzyl-B-D-rib furanosyl)thia~ole-4-carboxamide (5O0 g, 8.31 mmol) in methanol (15 ml) was stirred with methanolic ammonia (satur-ated at 0C, 100 ml) in a pressure bottle at room temperature for 2 days. The solvent was evaporated and the residue wa~
chromatographed through a column (2.5 x 35 cm3 o silica gel (100 g) packed in ethyl acetate. E-~ution of the column with a solvent system (eth~l acetate-l propanol-water, 4:1:2; v~v;
top layer3 removed tne fast-moving methyl benzo~te and benz-amide~ The slower moving, major, W and sugar-positive fractions were collected and the solvent was evaporated in vacuo. The residue (syrup), thus obtained, was readily crys-tallized from e-th~nol-ethyl ace-tate to provide 106 g (74%~
of pure product, compound 1: mp 144-145 C.; la~ Sp-1'I.3 ( 1 DMF); W ~ pHl 237 nm (8640); UV ~maXP
H ~R(Me2SOd~)~7.5 7.8 [S(br),2,CONH2]~ Me2SO-d6-D2O~-
4.g9(d,1,J=5 HZr Hle) t 8.25 ~s,ltH5)~ Anal. ~CgHl~N2O5S) C,H,N,S.
2~(2,3~5-Tri-O-Acetyl~-D-Ribofuranosyl3-thiazole-4-Carhoxamide, COMPOUND 2 Acetic anhydride (2.G ml) was added to an ice-cold solution of compound 1 (1.04 g, 4 mmol) in anhydrous pyridine tl6 m]) and the reaction solution was stirred at room temperature for 17 h. The solvent was evaporated in vacuo, the residue was dissolved in ethyl acetate, and the solution was washed with water and dried (MgSO4) The ethyl acetate portion was e~aporated in vacuo and the resi~ue thus obtained was crys~alized from water to provide 1~4 g (90%~ o~
compound 2 as white needles; mp 103C;lH NMR ~CDC13) 2.1 (3 s, 9,tri-O-acetyl), 6.2 and 7.15 [pair of s~br), 2, CONH2J, 8-2 (s, 1, H5)- Anal- (ClsHl~N2O8s) C~H~N~S-EXP~IPLE 3 2-(5-O-Phosphoryl-~-D~Ribofuranosyl)thiazole-4-Carboxamide ~2-~-D-Ribofuranosylthiazole-4-Carboxamide 5'~Phosphate), Water (151 mg, 8.4 mmol) was added carefully to a solution (maintained at 0C with stirring) of freshly dis-tilled phosphoryl chloride (2.0 g, 13.2 mmol), pyridine ~1~21 g, 14,4 mrnol) and acetonitrile (2 3 g, 56~7 mmol) 2-~-D--Ribofuranosy]-thiazole-4-caI^bo~lmide, compound 1, (clri~d over P2O5 arld powdered, 800 mg~ :3.0 mmol) was added t~ the solution and the xeaction mixture was stirred ~ontinuously for ~l hrs. at 0C. The reaction mixture was poured into ice water ~ca. 50 ml) and the pH was adjusted to 2.0 with 2N
sodium hydroxide. The solu-~ion was applied to a col~nn o~
activated charcoal (20 g3, and the column was washed thoroughly with water until t~le elua~e was salt~free. The column was eluted with a solution ethanol-water concentrated ammonlum hydroxide (10^10:1) and the fractions (25 ml each) were collected. The fractions containing pure (tlc, silica gel, ace-tonitrile~0.1 N ammonium chloride (7:3)~ nucleotide, com-pound 3, were collected and evaporated to dryn(ess under vacuum. The anhydrous residue was dissolved in water and passed through a column of Dowex* 50-W-X8 120-50 mesh, H form, 15 ml). The column was washed with water and the fraction containing the nucleotide was collected. The solution was concentrated to a small volume (5 ml) and passed through a column of Dowex* 50W-X8 (20 ~ 50 mesh, Na form, 15 ml). The column was washed with water. The nucleotide containing fraction was lyophilized. The residue was triturated with ethanol, collected by filtration and dried (P2O5)~ to provide 560 mg (47~) of compound 3 as monosodium dihy~rate in the crystalline form.
AnaL. calcd~ for CgH]2N~O8PSNa-2H~O: C, 27~13;
H, 4.0g; N,7~04; P, 7.78; S, $.05. Found: C, 27~42; ~I, 3.87;
N, 7~07i P, 8.03; S, 8.41.
*trade mark - 13 -^
mab/ ~' 4I'~
2-(5-O-Acetyl~-D-Ribofllranosyl)thiazole-4-Caxboxamide A solution of 2~(~,3-O-isopropylidene-~-D-ribo--furanosyl)thiazole-4-carboxamide 11.5 g, 5 mmol) (prepared as per Fuertes, et al; ~. Org. Chem., Volume 41, NO~ 26, 1976, 4074) in anhydrous pyridine (2Q ml) was chilled in an ice~
water bath and acetic anhydride (2.5 ml) was slowly added with stirring. The reaction solukion was allowed to warm to roo~ temperature and stirriny was continued for 15 h. The solvent was evaporated in vacuo and the residue was dissolved in ethyl acetate and washed with water. The ethyl acetate portion was evaporated in vacuo and the residue was dissolved in 80% acetic acid ~25 ml). The solution was heate~ on a steam bath for 30 mins. and the solvent evaporated in ~acuo. The residue was dissolved in ethyl acetate, washed once with water and dried over MgSO4. The ethyl acetate portiorl was evaporated and the crude product was passed through a column of silica gel ~100 g, packed in chloroform) and eluted with 20~ (V/V) ethyl acetate in chloroEormr The nucleoside bearing fractions were pooled and evaporated to yield 1~05 g (70%~
o compound 4. (CllH14N2O6S~o As illus-trative examples of the use of compound 1 and other illustrative compounds of the invention 3 examples
2~(2,3~5-Tri-O-Acetyl~-D-Ribofuranosyl3-thiazole-4-Carhoxamide, COMPOUND 2 Acetic anhydride (2.G ml) was added to an ice-cold solution of compound 1 (1.04 g, 4 mmol) in anhydrous pyridine tl6 m]) and the reaction solution was stirred at room temperature for 17 h. The solvent was evaporated in vacuo, the residue was dissolved in ethyl acetate, and the solution was washed with water and dried (MgSO4) The ethyl acetate portion was e~aporated in vacuo and the resi~ue thus obtained was crys~alized from water to provide 1~4 g (90%~ o~
compound 2 as white needles; mp 103C;lH NMR ~CDC13) 2.1 (3 s, 9,tri-O-acetyl), 6.2 and 7.15 [pair of s~br), 2, CONH2J, 8-2 (s, 1, H5)- Anal- (ClsHl~N2O8s) C~H~N~S-EXP~IPLE 3 2-(5-O-Phosphoryl-~-D~Ribofuranosyl)thiazole-4-Carboxamide ~2-~-D-Ribofuranosylthiazole-4-Carboxamide 5'~Phosphate), Water (151 mg, 8.4 mmol) was added carefully to a solution (maintained at 0C with stirring) of freshly dis-tilled phosphoryl chloride (2.0 g, 13.2 mmol), pyridine ~1~21 g, 14,4 mrnol) and acetonitrile (2 3 g, 56~7 mmol) 2-~-D--Ribofuranosy]-thiazole-4-caI^bo~lmide, compound 1, (clri~d over P2O5 arld powdered, 800 mg~ :3.0 mmol) was added t~ the solution and the xeaction mixture was stirred ~ontinuously for ~l hrs. at 0C. The reaction mixture was poured into ice water ~ca. 50 ml) and the pH was adjusted to 2.0 with 2N
sodium hydroxide. The solu-~ion was applied to a col~nn o~
activated charcoal (20 g3, and the column was washed thoroughly with water until t~le elua~e was salt~free. The column was eluted with a solution ethanol-water concentrated ammonlum hydroxide (10^10:1) and the fractions (25 ml each) were collected. The fractions containing pure (tlc, silica gel, ace-tonitrile~0.1 N ammonium chloride (7:3)~ nucleotide, com-pound 3, were collected and evaporated to dryn(ess under vacuum. The anhydrous residue was dissolved in water and passed through a column of Dowex* 50-W-X8 120-50 mesh, H form, 15 ml). The column was washed with water and the fraction containing the nucleotide was collected. The solution was concentrated to a small volume (5 ml) and passed through a column of Dowex* 50W-X8 (20 ~ 50 mesh, Na form, 15 ml). The column was washed with water. The nucleotide containing fraction was lyophilized. The residue was triturated with ethanol, collected by filtration and dried (P2O5)~ to provide 560 mg (47~) of compound 3 as monosodium dihy~rate in the crystalline form.
AnaL. calcd~ for CgH]2N~O8PSNa-2H~O: C, 27~13;
H, 4.0g; N,7~04; P, 7.78; S, $.05. Found: C, 27~42; ~I, 3.87;
N, 7~07i P, 8.03; S, 8.41.
*trade mark - 13 -^
mab/ ~' 4I'~
2-(5-O-Acetyl~-D-Ribofllranosyl)thiazole-4-Caxboxamide A solution of 2~(~,3-O-isopropylidene-~-D-ribo--furanosyl)thiazole-4-carboxamide 11.5 g, 5 mmol) (prepared as per Fuertes, et al; ~. Org. Chem., Volume 41, NO~ 26, 1976, 4074) in anhydrous pyridine (2Q ml) was chilled in an ice~
water bath and acetic anhydride (2.5 ml) was slowly added with stirring. The reaction solukion was allowed to warm to roo~ temperature and stirriny was continued for 15 h. The solvent was evaporated in vacuo and the residue was dissolved in ethyl acetate and washed with water. The ethyl acetate portion was evaporated in vacuo and the residue was dissolved in 80% acetic acid ~25 ml). The solution was heate~ on a steam bath for 30 mins. and the solvent evaporated in ~acuo. The residue was dissolved in ethyl acetate, washed once with water and dried over MgSO4. The ethyl acetate portiorl was evaporated and the crude product was passed through a column of silica gel ~100 g, packed in chloroform) and eluted with 20~ (V/V) ethyl acetate in chloroEormr The nucleoside bearing fractions were pooled and evaporated to yield 1~05 g (70%~
o compound 4. (CllH14N2O6S~o As illus-trative examples of the use of compound 1 and other illustrative compounds of the invention 3 examples
5 through 12~ below, are given. In these examples~ the efficacy of the compounds is demonstrated using the standaxd tests against cextain malignant tumors. The tesks utilized in these illu~trative examples were conducted by the Develop-mental ~'herapeutics Program, Division of Cancer Treat~ent, Nakîonal Cancer Institute. The tests were supervised by this ~4 --agency utiliz:ing the:ir standarcl protocols and procedures. All tests conformed to these protoco:Ls and all tests were evaluated under -the criteria defined by these protocols. I'he fol]owin~
representative examples illustrate confirmed activity of the illustrative compounds of the in~ention against screenin~
tumor systems of the National Cancer Institute.
For purposes of the Eollowing examples, the abbre-viation IP stands for intraperitoneal and IV stands for intra-venousO The mean and median survival times are calculated in instruction 14 (revised 6/78) of the Screening Data Su~mary9 Devel3pmental Therapeutics Prog~am, Division of ~ancer '~reat-ment, National Cançer Ins-titute. The contents of this Screening Data Summ,ary including appropriate revisions are herein incorporated by ref~rence.
In the illustrated examples below, the vehicle used as carrier for the drug was, injected (minus any drug therein) into the control animals at the same level of use o~
the yehicle in the drug treated animals in order to eliminate any vehicle effect of the tests.
As an indicator of reproducible activity, compou~d 1 of the invention was screened against L-1210 lymphoid leuke mia in yivo using CD2Fl male mice as the testing species.
The parameter o~ efficacy chosen was based upon the median suxvival time of the animals treated wi-th the drug vs. appr~-priate control group animals. Both drug treated animals and control ~roup animals were inoculated IP ~itA 10 seed cells o~ 1210 lymphoid leukemia in Ascitic fluid~
One day after tumor inoculation, the drug ~roup -- 15 ~ ~ ~ a a,t of animals was pl~ced on a reqimell of trecltrnent of compound 1 at the dose levels as no-ted below, table 1. The druy tr~ated group of animals was inocula-ted once daily for five days at the doses noted by IP injection of the test compound appro-priately d-luted with water.
Day six was chosen as an indication of druy toxicity. In this example, all drug treated animals survi~;ed - through day six. Death of drug treated ani~als after day six was, there~ore, attributed to tumor death$ and n~t d~u~
1~t~xi~i`ty~
The median death day of the control group was day 8~5. As noted in table 1 below, the median death day of the dru~ treated group was longer at all levels of drug tested and was si~nificantly longer at greater than 50 mg/Kg (amount of drug/wei~ht of test animal). The results shown in table 1 below indicate that in this multiple dose assay, t~e drug showed positive activity. A percent of drug treated animals/-control anin)als greater than 1~5% is taken as positive drug activity.
TABI,E 1 Percent Dru~ DoseTreated Group Control Group Treated ~ni~als/
mg~KgSurvi~val Time Survival Time Control Animals 200 14O3 8,5 168%
100 12.~ 1~9%
11~0 129%
10.2 1~%
12 7 5 9.5 111%
Compound2r 2-(2~3,5 t:ri-O-ace-tyl-~D-ribofurano-syl)thiazo]e-4-carboxamide was screened in a manner similar to that shown in Example 5, howeve.r, ~he tumor system used as the test tumor was P388 lymphocytic leukemia. 106 seed cells were used to initiate the tumor in bo-th the control group and the drug treated group of animals. The same strain of ~ice was used exoept female mice were substituted for male~. Test results were based on mean survival time and are expressed as T/C percentages (treated ani.mals~contxol animals~
as per Example 5.
In the drug treated animals, treatmen~ was initiated one day after tumor inocu]ation and the drug was given at the dose levels noted below in table 2. Drug treat-ment was fo:r nine days and drug toxici.ty, as in Example 5, was measured on day six. At the 100 mg~Kg level, one animal did not survive the toxicity cut-off date.
Average day of death for the control group was 10.2 days whereas at the lowes-t level of drug treatment -the ~0 treated animals su.rYived or more than 15 days. As with Ex~mple 5, 125% increase in longevity of treated animals over oontrol animals was taken as indicative of positive drug response.
TABI,E 2 Peroent Drug Dose Treated Group Control Group Trea-ted Animals/
mg/Kg Survival Time Survival Time Control Animals 2~0 1~.3 10.2 17~%
1~0 18.0 176~
15.3 150%
mab/~ll Compound 1 is al.so indica-ted as being active agains-t P38g lymphocy-tic leu~emia as per Examples 6a, b, an~
c, ancl 2-(2,3,5-tri-0-acetyl. ~-D-ribofuranosyl)thiazole 4-carboxamide, compound 2, is indicated also as being active agains-t P388 lymphocyti.c leukemi.a as per Example 7. In both of these examples, the compound successfully passed the DN
2 ~Decision Network) criteria of the National Cancer Institute Testing. For examples 7 and 8, CD2Fl femaie mice were used and challenged with P388 lymphocytic leukemia tumors. The median survival time of the drug -treated ~nimals w~s compared to appropriate control animals and based on this criteria both of the compounds -tested were considered as active antitumor agents~ The test period was for thirty days in both Examples 7 and 8.
For Examples 7 and 8, as well as Examples 9 and 10 below, any animal of the d.rug treated group which survived beyond the end of the testing period was then evaluated and placed in one of three groups. The first group wa.s designated as cured, meaning the animal was successfully cured of the tumor. I'he second group designation was no--takes, meanins survival of the animal was considered to be due to failure of tumor implant. The remaining group was designated as tumor survivors meaning the animals lived beyond the test cut-off date but could not be classified as either cured or no-takes.
For both Examples ~ and 8, thirty anim~ls were used as -the control group and six animals each were used at each dose level indicated in tables 3 and 4 below in the dru~
treated groups. In bo-th Examples 7 and 8, for both the control group and the dru~ treated groups, tumors were induced - 18 ~
by IP i~oculation of -tumor seed cells on day ~ero followed by initiation of drug t~eatment orl day one. For bvth E~amples 7a and 8, sallne Wit~l -tween/80 was used as the dru~ vehicle.
~or Examples 7b and 7c, water was used as the druy vehicle.
In both ~he control grQUp and the drug treated group in Examples 7 and 8, the test animals were inoculated on day zero IP with 106 seed cells of P33B lymphocytio leukemia. In both Examples 7 and 8, treatment of the drug ~roup was started on day one and the drug was given IP once daily for nine days. Day six was utilized as the cut-off date for de~th attributable to toxicity of the drug. In only one i~nstance, in Example 7b, was animal mortality attribute~ to drug toxicity. Efflcacy of treatment was determined by comparing median survival time of the drug treated animals compared to median survival time of the control animals, and is expressed as percentage increase of treated animals/control animals (T/C1 as per Example 5.
EXAMPLE 7a In this example drug treated animals were injected IP with the dose level noted in Table 3 below. Six animals were treated at each dose level. No control animals survived beyond day 18 with a median death date of day 12~5~ The median death day of the drug treated animals is as shown in Table 3a below. At the 50 mg~Kg level, one drug treated animal survi~ed and was ~udged as a no-take.
EXAMPLE 7b This example was performed as per Example 7a at dos~ leYels as noted in Table 3b below A sur~ivor at bot~
the 700 and 800 my~Kg level was judged as cured~ No controls ~ 19 -Iilab/~ ,` t~Q~
survived beyond day :L2 and the mean death day of -the control group was day 11.
E~AMPLE 7c This example was run as per Example 7a above at dose levels noted in Table 3c be:Low. All controls were dead ~y day 14 with a mean death date of day 11.9~ At the 500 mg/Kg level, one animal was judged as a cure.
Compound 2, 2-(2 ! 3,5-tri-0-acetyl ~-D-ribofuranosyl~-thiazole-4-carboxamide was tested as per Example 7a above at dose levels noted in Table 4 below. No controls survived be-yond day lB with an average death date of day 12.6. At the 50 mg/Kg level, one surv~`ving animal was judged as a no take.
Both compounds 1 and 2 are indicated as being active antitumor agents in the multiple dose studies noted in Examples 7 and 8.
TABLE 3a Percen-t Drug Dose Treated Group Control Group Treated Animals/
mg/Kg Sur~ival Time Survival Time Control Animals 400 20.3 12.6 161%
200 1~.0 150%
100 18~3 1~5%
15.3 121%
~5 14.3 ~13%
12.5 13.9 110%
TABLE 3b Percen~
Drug Dose Treated Group Control Group Treated Anima~s/
mg/Kg Survival Time Survival Time Control Animals 800 2700 11.~ 2~5%
700 ~7.0 2~5%
~00 2S,8 ~3~%
500 21.~ 190%
400 24.7 2~4 300 21.8 198 -- :20 --mctb/~
~ ~3~ ~t7~ ~
TABLE 3c Perc~nt Dr-lg Dose Treated Group Control Group Trea~ed Animals/
mg/Kg Survival Time Survival Time Con-trol ~nimals 800 11.8 ~1.9 99~
700 lOo ~36~i 600 28 ~ 3 ;23~%
5G0 25~0 210~
40G 24.0 201%
3~0 2300 lg3 Percent Drug Dose Treated Group Control Group Treated Animals/
mg/Kg Survival Time Survival Time Con~rol Animals 400 2~.3 1~.6 161 1~ 200 19~0 150 lOQ 1~.3 1~5%
15.3 1~1%
14.3 113%
12.5 13.7 ~10~
Compound 1 is indicated as being active against L-1210 lymphoid leukemia as per Example 9, and successfully passed the DN 2 criteria of the National Cancer Institute testing. For Example 9a and 9b, CD2Fl male mi.ce were used and challenged with L-1210 lymphoid leukemia. The mean survival ~ime of the test animals was co~pared to appropriate control animals and based on this cri.teria, compound 1 was considered as an active an~itumor agent. The test period was for thirty days. Test results are expressed as T/C as per Example 5.
In Examp].e 9a, 24 control animals were used and six test animals at each drug dose level as is indicated below in table 9a. I.n ~xample 9b, forty control animals were used and ten test animals each at drug dose levels as shown in table 9b helow. For both the control ~roup and the drug test group, tumo~s were induced by IP inoculation of tumor seed ~ells on day zexo followed by initiation of drug tre~tment on day one. For Example 9a, water was used as -the druc~ vehicle , .1 ~ 3~
and for Exarnple 9b saline was used as the druy vehicle In bo-th -the cont:rol ~roups and the dru~J treated groups in Examp1es 9a and 9b, the test animals were inoculated on day zero IP with 10 seed cel:Ls of L~1210 lymphoi.d leukemia.
.~or Example 9a~ drug -treatment was started on day one and cvmpound 1 given once daily Eor nine days. Day five was utilized as the cut-o~f date for death at-tributable to toxicity of the drug. In only one instance in Ex2mple 9~ was mortality attributed to drug toxicity r Efficacy of treatment was deter-mined by comparing mean survival time of drug treated animals with mean survival time of tne con-trol animals and is expressed as percentage increase of treated animals/control a~imals (T~C) as per Example 5.
EXAMPLE 9a In this example, the drug treated animals wer~
injected IP with dose levels noted in table 5a below. Six animals were treated at each dose levelO No control animals survived beyond day ten with a mean death date of day 9.7.
The mean death day of the drug treated animals is as shown in table 5a below.
EXAMPLE 9b In this example, druy treated animals were injected IP with the dose level noted in table 5b below. Ten animals were treated at each dose leYel. No control animals sur~ived beyond day ten with a mean death date of day 9Ø The mean death day of the treated animals is as shown in table Sb below.
TABLE 5a Percent Drug Do~e Trea-ted Group Control Group Treated Animals/
mg/Kg 5urvival Time Survival Time Colltrol Animals 400 lg.7 3.7 19~%
200 15.3 1~7%
100 1~.0 144%
13.2 136~
12.8 131%
T~BLE 5b Percent Drug Dose Treated Group Control ~roup Trea-ted Animals~
mg/KgSurvival Time Survival TimeControl Animals 800 15~4 ~.0 171%
600 20.7 ~30%
400 20.2 224%
2~0 16.4 182%
100 16~5 183%
Compound 1 is indicated as being active against I,ewis lung carcinoma as per ~xample 10 and successfully passed the DN 2 criteria of the National Cancer Institut~ Testing~
E'or Example 10, B6D~Fl male mice were used and challenged with Lewis lung carcinoma. The median survival time of the test animals was compared to appropriate control animals and based on this criteria compound 1 was considered as an effective antitumor agent.
In Exarnple 10, forty control animals were used and ten test animals each at dose levels indicated below in Ta~le
representative examples illustrate confirmed activity of the illustrative compounds of the in~ention against screenin~
tumor systems of the National Cancer Institute.
For purposes of the Eollowing examples, the abbre-viation IP stands for intraperitoneal and IV stands for intra-venousO The mean and median survival times are calculated in instruction 14 (revised 6/78) of the Screening Data Su~mary9 Devel3pmental Therapeutics Prog~am, Division of ~ancer '~reat-ment, National Cançer Ins-titute. The contents of this Screening Data Summ,ary including appropriate revisions are herein incorporated by ref~rence.
In the illustrated examples below, the vehicle used as carrier for the drug was, injected (minus any drug therein) into the control animals at the same level of use o~
the yehicle in the drug treated animals in order to eliminate any vehicle effect of the tests.
As an indicator of reproducible activity, compou~d 1 of the invention was screened against L-1210 lymphoid leuke mia in yivo using CD2Fl male mice as the testing species.
The parameter o~ efficacy chosen was based upon the median suxvival time of the animals treated wi-th the drug vs. appr~-priate control group animals. Both drug treated animals and control ~roup animals were inoculated IP ~itA 10 seed cells o~ 1210 lymphoid leukemia in Ascitic fluid~
One day after tumor inoculation, the drug ~roup -- 15 ~ ~ ~ a a,t of animals was pl~ced on a reqimell of trecltrnent of compound 1 at the dose levels as no-ted below, table 1. The druy tr~ated group of animals was inocula-ted once daily for five days at the doses noted by IP injection of the test compound appro-priately d-luted with water.
Day six was chosen as an indication of druy toxicity. In this example, all drug treated animals survi~;ed - through day six. Death of drug treated ani~als after day six was, there~ore, attributed to tumor death$ and n~t d~u~
1~t~xi~i`ty~
The median death day of the control group was day 8~5. As noted in table 1 below, the median death day of the dru~ treated group was longer at all levels of drug tested and was si~nificantly longer at greater than 50 mg/Kg (amount of drug/wei~ht of test animal). The results shown in table 1 below indicate that in this multiple dose assay, t~e drug showed positive activity. A percent of drug treated animals/-control anin)als greater than 1~5% is taken as positive drug activity.
TABI,E 1 Percent Dru~ DoseTreated Group Control Group Treated ~ni~als/
mg~KgSurvi~val Time Survival Time Control Animals 200 14O3 8,5 168%
100 12.~ 1~9%
11~0 129%
10.2 1~%
12 7 5 9.5 111%
Compound2r 2-(2~3,5 t:ri-O-ace-tyl-~D-ribofurano-syl)thiazo]e-4-carboxamide was screened in a manner similar to that shown in Example 5, howeve.r, ~he tumor system used as the test tumor was P388 lymphocytic leukemia. 106 seed cells were used to initiate the tumor in bo-th the control group and the drug treated group of animals. The same strain of ~ice was used exoept female mice were substituted for male~. Test results were based on mean survival time and are expressed as T/C percentages (treated ani.mals~contxol animals~
as per Example 5.
In the drug treated animals, treatmen~ was initiated one day after tumor inocu]ation and the drug was given at the dose levels noted below in table 2. Drug treat-ment was fo:r nine days and drug toxici.ty, as in Example 5, was measured on day six. At the 100 mg~Kg level, one animal did not survive the toxicity cut-off date.
Average day of death for the control group was 10.2 days whereas at the lowes-t level of drug treatment -the ~0 treated animals su.rYived or more than 15 days. As with Ex~mple 5, 125% increase in longevity of treated animals over oontrol animals was taken as indicative of positive drug response.
TABI,E 2 Peroent Drug Dose Treated Group Control Group Trea-ted Animals/
mg/Kg Survival Time Survival Time Control Animals 2~0 1~.3 10.2 17~%
1~0 18.0 176~
15.3 150%
mab/~ll Compound 1 is al.so indica-ted as being active agains-t P38g lymphocy-tic leu~emia as per Examples 6a, b, an~
c, ancl 2-(2,3,5-tri-0-acetyl. ~-D-ribofuranosyl)thiazole 4-carboxamide, compound 2, is indicated also as being active agains-t P388 lymphocyti.c leukemi.a as per Example 7. In both of these examples, the compound successfully passed the DN
2 ~Decision Network) criteria of the National Cancer Institute Testing. For examples 7 and 8, CD2Fl femaie mice were used and challenged with P388 lymphocytic leukemia tumors. The median survival time of the drug -treated ~nimals w~s compared to appropriate control animals and based on this criteria both of the compounds -tested were considered as active antitumor agents~ The test period was for thirty days in both Examples 7 and 8.
For Examples 7 and 8, as well as Examples 9 and 10 below, any animal of the d.rug treated group which survived beyond the end of the testing period was then evaluated and placed in one of three groups. The first group wa.s designated as cured, meaning the animal was successfully cured of the tumor. I'he second group designation was no--takes, meanins survival of the animal was considered to be due to failure of tumor implant. The remaining group was designated as tumor survivors meaning the animals lived beyond the test cut-off date but could not be classified as either cured or no-takes.
For both Examples ~ and 8, thirty anim~ls were used as -the control group and six animals each were used at each dose level indicated in tables 3 and 4 below in the dru~
treated groups. In bo-th Examples 7 and 8, for both the control group and the dru~ treated groups, tumors were induced - 18 ~
by IP i~oculation of -tumor seed cells on day ~ero followed by initiation of drug t~eatment orl day one. For bvth E~amples 7a and 8, sallne Wit~l -tween/80 was used as the dru~ vehicle.
~or Examples 7b and 7c, water was used as the druy vehicle.
In both ~he control grQUp and the drug treated group in Examples 7 and 8, the test animals were inoculated on day zero IP with 106 seed cells of P33B lymphocytio leukemia. In both Examples 7 and 8, treatment of the drug ~roup was started on day one and the drug was given IP once daily for nine days. Day six was utilized as the cut-off date for de~th attributable to toxicity of the drug. In only one i~nstance, in Example 7b, was animal mortality attribute~ to drug toxicity. Efflcacy of treatment was determined by comparing median survival time of the drug treated animals compared to median survival time of the control animals, and is expressed as percentage increase of treated animals/control animals (T/C1 as per Example 5.
EXAMPLE 7a In this example drug treated animals were injected IP with the dose level noted in Table 3 below. Six animals were treated at each dose level. No control animals survived beyond day 18 with a median death date of day 12~5~ The median death day of the drug treated animals is as shown in Table 3a below. At the 50 mg~Kg level, one drug treated animal survi~ed and was ~udged as a no-take.
EXAMPLE 7b This example was performed as per Example 7a at dos~ leYels as noted in Table 3b below A sur~ivor at bot~
the 700 and 800 my~Kg level was judged as cured~ No controls ~ 19 -Iilab/~ ,` t~Q~
survived beyond day :L2 and the mean death day of -the control group was day 11.
E~AMPLE 7c This example was run as per Example 7a above at dose levels noted in Table 3c be:Low. All controls were dead ~y day 14 with a mean death date of day 11.9~ At the 500 mg/Kg level, one animal was judged as a cure.
Compound 2, 2-(2 ! 3,5-tri-0-acetyl ~-D-ribofuranosyl~-thiazole-4-carboxamide was tested as per Example 7a above at dose levels noted in Table 4 below. No controls survived be-yond day lB with an average death date of day 12.6. At the 50 mg/Kg level, one surv~`ving animal was judged as a no take.
Both compounds 1 and 2 are indicated as being active antitumor agents in the multiple dose studies noted in Examples 7 and 8.
TABLE 3a Percen-t Drug Dose Treated Group Control Group Treated Animals/
mg/Kg Sur~ival Time Survival Time Control Animals 400 20.3 12.6 161%
200 1~.0 150%
100 18~3 1~5%
15.3 121%
~5 14.3 ~13%
12.5 13.9 110%
TABLE 3b Percen~
Drug Dose Treated Group Control Group Treated Anima~s/
mg/Kg Survival Time Survival Time Control Animals 800 2700 11.~ 2~5%
700 ~7.0 2~5%
~00 2S,8 ~3~%
500 21.~ 190%
400 24.7 2~4 300 21.8 198 -- :20 --mctb/~
~ ~3~ ~t7~ ~
TABLE 3c Perc~nt Dr-lg Dose Treated Group Control Group Trea~ed Animals/
mg/Kg Survival Time Survival Time Con-trol ~nimals 800 11.8 ~1.9 99~
700 lOo ~36~i 600 28 ~ 3 ;23~%
5G0 25~0 210~
40G 24.0 201%
3~0 2300 lg3 Percent Drug Dose Treated Group Control Group Treated Animals/
mg/Kg Survival Time Survival Time Con~rol Animals 400 2~.3 1~.6 161 1~ 200 19~0 150 lOQ 1~.3 1~5%
15.3 1~1%
14.3 113%
12.5 13.7 ~10~
Compound 1 is indicated as being active against L-1210 lymphoid leukemia as per Example 9, and successfully passed the DN 2 criteria of the National Cancer Institute testing. For Example 9a and 9b, CD2Fl male mi.ce were used and challenged with L-1210 lymphoid leukemia. The mean survival ~ime of the test animals was co~pared to appropriate control animals and based on this cri.teria, compound 1 was considered as an active an~itumor agent. The test period was for thirty days. Test results are expressed as T/C as per Example 5.
In Examp].e 9a, 24 control animals were used and six test animals at each drug dose level as is indicated below in table 9a. I.n ~xample 9b, forty control animals were used and ten test animals each at drug dose levels as shown in table 9b helow. For both the control ~roup and the drug test group, tumo~s were induced by IP inoculation of tumor seed ~ells on day zexo followed by initiation of drug tre~tment on day one. For Example 9a, water was used as -the druc~ vehicle , .1 ~ 3~
and for Exarnple 9b saline was used as the druy vehicle In bo-th -the cont:rol ~roups and the dru~J treated groups in Examp1es 9a and 9b, the test animals were inoculated on day zero IP with 10 seed cel:Ls of L~1210 lymphoi.d leukemia.
.~or Example 9a~ drug -treatment was started on day one and cvmpound 1 given once daily Eor nine days. Day five was utilized as the cut-o~f date for death at-tributable to toxicity of the drug. In only one instance in Ex2mple 9~ was mortality attributed to drug toxicity r Efficacy of treatment was deter-mined by comparing mean survival time of drug treated animals with mean survival time of tne con-trol animals and is expressed as percentage increase of treated animals/control a~imals (T~C) as per Example 5.
EXAMPLE 9a In this example, the drug treated animals wer~
injected IP with dose levels noted in table 5a below. Six animals were treated at each dose levelO No control animals survived beyond day ten with a mean death date of day 9.7.
The mean death day of the drug treated animals is as shown in table 5a below.
EXAMPLE 9b In this example, druy treated animals were injected IP with the dose level noted in table 5b below. Ten animals were treated at each dose leYel. No control animals sur~ived beyond day ten with a mean death date of day 9Ø The mean death day of the treated animals is as shown in table Sb below.
TABLE 5a Percent Drug Do~e Trea-ted Group Control Group Treated Animals/
mg/Kg 5urvival Time Survival Time Colltrol Animals 400 lg.7 3.7 19~%
200 15.3 1~7%
100 1~.0 144%
13.2 136~
12.8 131%
T~BLE 5b Percent Drug Dose Treated Group Control ~roup Trea-ted Animals~
mg/KgSurvival Time Survival TimeControl Animals 800 15~4 ~.0 171%
600 20.7 ~30%
400 20.2 224%
2~0 16.4 182%
100 16~5 183%
Compound 1 is indicated as being active against I,ewis lung carcinoma as per ~xample 10 and successfully passed the DN 2 criteria of the National Cancer Institut~ Testing~
E'or Example 10, B6D~Fl male mice were used and challenged with Lewis lung carcinoma. The median survival time of the test animals was compared to appropriate control animals and based on this criteria compound 1 was considered as an effective antitumor agent.
In Exarnple 10, forty control animals were used and ten test animals each at dose levels indicated below in Ta~le
6. For both the control group and the drug treated group~
tu~oxs were induced by IV in~ection on day zero followed by initiating of drug treatment on day one. For Example 10 water was used as the drug ~ehicle.
In both the control group and the drug treated ~roup in Example 10, the animals were inoculated on day ~ero with a homo~enate of 10 seed cells of Lewis lung carcinoma.
'For Example 10, drug treatment was started on day one and compound 1 gi-ven once daily for nine days. Day ~ive was utilized as the cut-off date for deaths attributable to toxicity of thP dru~. There was no mortality attributable to drug toxicity in this example~ Efficacy of treatment was de-termined by comparing median survlval time of dru~ treated animals with mab~',' ~ 3,~Ji~
median survival time of the control animals and is expressed as pe~rcentac3e increase of treatecl animals/con-trol animals (T/C) as per Example 5.
The test period was for sixty days and ~t the end of the sixty day perivd all animals surviving in the test yroups were evaluated as either cured, no-takes~ or tumor sur-vivors as per Example 5 a~ove.
In th.is example, the drug treat.ed animals were injected IP with the dose level noted in table 6 below. Ten animals were treated at each dose level. No control animals survived beyond day 23 with a medi.an death date of day 18.4.
At the test levels of 400, 200 and 25 mg/Kg all test animals survive~ tIIe s xty day test period~ Because of this f.~ct, the T/C ratio noted in table 6 below is a constant figure based on assigning survival day rate of sixty to the treated animal.s and a median death date of 18.4 days to the control animals~
In Example 10 at both the 200 and 400 my~Kg level, all ten surviving test animals were judged as cured. At the 100 mg/Kg level, there were eight cures and one tumor survivor with one death noted on day 46. At the 50 mg/Kg level, there were nilIe cures and one death on day 47.
Compound 1 i.s indicated as being an active antitumor agent in the multiple dose studies noted in Example 1 Percent ~rug Dose Treated Group Control Group Treated Animals~
mg/Kg Survival Time Survival Time Control Animals ~00 60 18.~ 326%
20~ 60 326%
100 60 32~%
326~
3~%
As is shown in Example 10 above, compound 1 shows outstanding activity against Lewis l.ung carcinoma. Lewis lun~
carcinoma i~ an excellent example of a metastatic tumor system. The tests and control animals of Example 10 were inoculated 1~ with a homogenate of the -tumor. Dramatic expres-sion oE this tumor is then seen in the lungs. As was noted - ~4 -mab/l~
p.revi.ous:Ly, ~he ahil:ity to m~tastasize is a property -that uni.q~lely charac-terizes a n~alignan-t tu~or f.rom a beniyn tumor. In Example 10, n~t only was -~he median survival time of d~lg treated animals dralratically extended but, a-t the cessation of the -test period, except at one dose level, at least 80% cures w~re no-ted and at two of those levels lOQ~ cures were present.
:~X~MELE 11 Compound 3~ 2-(5-O-phosphoryl-~-D-riboEuranosyl~thia~ole-4-carboxamide is i~licated as being active against L-1210 Lymphoid leukemia æ per e~mnple lla and llb. These exc~mples were per~ormed essentiall~
as per exa~ple 9 c~bove except as noted. T~e compourld test ~osage levels axe as no-ted in tables 7a and 7b belcw Eor Example lla a~d llb respec-tive7y.
In these exlmples, ~hirty-six control an~als wexe used and six tes~ ~n~m~
each at the drug dose levels as is shawn in tables 7a and 7b~ Saline was used as the drug vehicle. No drug -toxicity was nobed in the test ~ni~ for either exampl~s lla or llb. No control ~n;~l~ survived beyond day 10 in example lL~ with a mean death date of day 8~3 ~nd ~eyond day ele~en in ~x~rle llb with a mean death date of day 10~1.
For bo~h the control groups and the drug test gro~sg tu~rs were ;n~ur.~ by IP inocula-tion of tumor seed cells on day 2ero follcwed by i~ tiation of drug -trea~ ~ t on day one wherein compolmd 3 was given once daily for 5 days. Test results are expressed as T/C as per example 5.
TABLE 7a Percent Dl'Ug Dose Treated Group Control Group Treated Animals/
mg/Kg Survival Time Survival Time Con-trol Animals 80~ 16.3 ~.3 196%
400 15~2 183%
200 21.5 259%
100 15.5 1~6%
11.3 136%
TABLE 7b Percent Dru~ Dose Treated Group Control Group Treated Animals/
mg/Kg Sur~i~al Time Survival Time Control Animals 6~Q 16.2 10.1 160%
400 15.7 15~%
~00 14.7 145%
100 12.3 121~
13.0 128%
~or Example 12, co~pound 1 was given IP to a ~roup of AKD2F~ r~ce which were i~flicted by intercra~ial inoculation with L~;s ..~, rnah/, ~
~ 7,~
l~mg seed cells to est~ lish brain tu~.ors. The resul-ts in Table 8 bel~w indicate that the IP inoculation of the afflic-ted ani~ls with cQmpound 1 resulted in reduction of the brain tumors indicating succ,essful crossing of the blood brain barrier by compound 1 follawing the IP in~ection lnto the afflic-ted allLmals~
For this -test, 32 control An;mAl~ were used and no control ~n;~ . survived beyond day eleven with a mean death date of da~ 9.6 for the controls~ Eight test ~ni~r~l~ were used for each drug dose le~el with the exception of the 300 mg/Kg level as is shown in table 8 ~elow~ Water wa5 used as the drug vehicle. For both the control ~roup and ~he -test gro~p~ .~3rs were induced on day zeno with i~itiation cf dru~ treatment on day one where compoun~ 1 was given once daily ~or nin2 days. Test results are expressed as T/C as per example 5.
Percent Drug Dose Treated Group ContrQl Group Treated Animalsf mg~Kg Survival Time Survival Time Control Animals 800 21O3 9.6 221%
700 20.3 211%
600 20.3 211%
~0 5 213%
150 20.5 197%
~5 19.0 185%
37.5 16.0 1~6%
25.0 16.6 172%
In a significant number of disease states of the brain of both pa~h~nic and host dysfunction origins, treatment is inhibited b~
the lack of drug transfer across the blood brain barrier. In. certain in-stances wherein appropriate -treatment of a disease state is k2~0wn, com~
plications can arise in treating these diseases when they are located intercra~ially because of the lack of ~ransfer across ~he blocd brain b~rrier of effective concentrations of appropriate che~otheraputic agen~s.
The indication, as seen in table 8, that coI~?~und 1 success~ ~ly crosses t~e blood brain barrier i5 thus very prorrlising for the trea-bment of brain h ~
DJ~
t~nors.
~ he :Eollcwinc3 representat:i,ve e,~ mples, 13 throuyh 17, are gi.ven for the fo~ lations of the active csmpo~md of the inven-ti.on in illustrative pharmaceutical compositions utiliziny illustrative carriers~
In these examples~ ex~nple 13 illustrat:es the use of the compou~lds of the inven-tion in injectables suitable for intra~enous or o~he~ type5 of injec-tion into the host anLmaL. Example 14 is directed to an oral syrup prepar-ation. Example 15 to an oral capsule p.reparation and E~3mple 16 to oral tablets. Exa~ple 17 is di.r.ec-t~d to use of the compoun~s of the invention in suitable suppositories. For Examples 13 through 17 the in~redients a~e listed follcwed by ~he me-thods of preparing the co~çc~ition.
E~MPLE 13 INJECTABLES
Example 13a Csmpound 1 Qo~pound 1 250mg - 1000 mg Water ~or Injectio~ USP q.s.
Cc~npound 1 is dissolved in the water and passecl through a 0.22 ~ filter. Ihe -filtered solution is added to ampoules or vials, sealed and sterilized.
Example 13b Compound 3 Canpound 3 as a Sodium Salt 250 mg - 1000 mg Water for Injection USP q.s.
~repared as per Example 3a, above.
E~AMPLE 14 SYRUP
Example 14a Compound 1 250 mg Active ingredient/5 ml syrup Comp~l~nd 1 sOg Purified Water USP 200 ml Cherry Syr~p ~s. or lOOO ml Cc~npow1d 1 is clissolved in the wa-ter and to ~his solu~ion the syrup is added with mild s-tirring.
Example 14b Compound 3 250 mg Ac-tive ingred:Lerlt/5 ml syrup Compound 3 as a Sodium Salt 50.0 g Purified Water USP q.s. or 200 ml Cherry Syrup q.s. ad 1000 ml Prepared as per Example 14a above.
CAPSULES
Example 15a Compound 1 100 mg, 250 mg or 500 mg Compound 1 50U g Lactose USP, Anhydrous qOs. or 200 g Stero-tex* Powder HM 5 g Combine compound 1 and the Lactose in a t~in-shell blender equipped with an intensifier bar~ Tumble ble~d for two minutes, Eollowed by blending for one minute with ~he intensifier bar and then tumble blend again for one mi~ute. A
port~on of the blend i~s then mixed with the Sterotex* ~owder, passed through a #30 screen and added back to the rema~Lnder o~
the blend. The mixed ingredients are then blended for one minute, blended with -the intensifier bar for thirty seconds and tumble bl.ended for an additional minute Appropri~te si~ed capsules are filled with 141 mg, 352 5 mg or 705 mg of the blend, respectively~ for the 100 mg, 250 mg an~ 500 mg con~
taining capsules.
EXAMPLE 15b Example 15b Compound 2 100 mg, 250 mg or 500 mg Compound 2 *trade marJc 28 -3~
I,actose USP, Anhydrous q.s. or 200 g Sterotex* Powder ~IM 5 ~J
Mix and fill as per Example 15 a.
Example 15c Compound 4 100 mg, 250 mg or 500 mg Compound 4 5~0 g Lactose USP~ ~nhydrous q.sO or 20Q g 5terotex* Powder HM 5 g Mix and fill as per Example 15a EX~MPLE 16 TABLETS
Example 16a Compound 1 100 mgt 200 mg or 500 mg Compound 1 500 g Corn Starch NF 200 g Cellullose, Microcrystalline 46.0 g Sterotex* Powder HM 4.0 g Purified Water q.s. or 300 ml Combine the corn starch, the cellulose and Compoun~
1 -together .in a planetary mixer and mix for two minutes~ Add the water to this combination and mix for one minute. The resulting mix is spread on trays and dried in a hot air oven at 50C until a moisture level of 1 to ? percent is o~tained.
The dried mix is then milled with a Fitzmill through a ~RH2B
screen at medium speed. The Sterotex~ Powder is adde~ to a po~tion o~ the mix and passed through a #30 screen, and added ~ack to the milled mixture and the total blended for five minutes by drum rol.lincJ. Compressed tablets oE 150 m~, 375 mcJ
~ 29 -*trade mark rn;~ 3
tu~oxs were induced by IV in~ection on day zero followed by initiating of drug treatment on day one. For Example 10 water was used as the drug ~ehicle.
In both the control group and the drug treated ~roup in Example 10, the animals were inoculated on day ~ero with a homo~enate of 10 seed cells of Lewis lung carcinoma.
'For Example 10, drug treatment was started on day one and compound 1 gi-ven once daily for nine days. Day ~ive was utilized as the cut-off date for deaths attributable to toxicity of thP dru~. There was no mortality attributable to drug toxicity in this example~ Efficacy of treatment was de-termined by comparing median survlval time of dru~ treated animals with mab~',' ~ 3,~Ji~
median survival time of the control animals and is expressed as pe~rcentac3e increase of treatecl animals/con-trol animals (T/C) as per Example 5.
The test period was for sixty days and ~t the end of the sixty day perivd all animals surviving in the test yroups were evaluated as either cured, no-takes~ or tumor sur-vivors as per Example 5 a~ove.
In th.is example, the drug treat.ed animals were injected IP with the dose level noted in table 6 below. Ten animals were treated at each dose level. No control animals survived beyond day 23 with a medi.an death date of day 18.4.
At the test levels of 400, 200 and 25 mg/Kg all test animals survive~ tIIe s xty day test period~ Because of this f.~ct, the T/C ratio noted in table 6 below is a constant figure based on assigning survival day rate of sixty to the treated animal.s and a median death date of 18.4 days to the control animals~
In Example 10 at both the 200 and 400 my~Kg level, all ten surviving test animals were judged as cured. At the 100 mg/Kg level, there were eight cures and one tumor survivor with one death noted on day 46. At the 50 mg/Kg level, there were nilIe cures and one death on day 47.
Compound 1 i.s indicated as being an active antitumor agent in the multiple dose studies noted in Example 1 Percent ~rug Dose Treated Group Control Group Treated Animals~
mg/Kg Survival Time Survival Time Control Animals ~00 60 18.~ 326%
20~ 60 326%
100 60 32~%
326~
3~%
As is shown in Example 10 above, compound 1 shows outstanding activity against Lewis l.ung carcinoma. Lewis lun~
carcinoma i~ an excellent example of a metastatic tumor system. The tests and control animals of Example 10 were inoculated 1~ with a homogenate of the -tumor. Dramatic expres-sion oE this tumor is then seen in the lungs. As was noted - ~4 -mab/l~
p.revi.ous:Ly, ~he ahil:ity to m~tastasize is a property -that uni.q~lely charac-terizes a n~alignan-t tu~or f.rom a beniyn tumor. In Example 10, n~t only was -~he median survival time of d~lg treated animals dralratically extended but, a-t the cessation of the -test period, except at one dose level, at least 80% cures w~re no-ted and at two of those levels lOQ~ cures were present.
:~X~MELE 11 Compound 3~ 2-(5-O-phosphoryl-~-D-riboEuranosyl~thia~ole-4-carboxamide is i~licated as being active against L-1210 Lymphoid leukemia æ per e~mnple lla and llb. These exc~mples were per~ormed essentiall~
as per exa~ple 9 c~bove except as noted. T~e compourld test ~osage levels axe as no-ted in tables 7a and 7b belcw Eor Example lla a~d llb respec-tive7y.
In these exlmples, ~hirty-six control an~als wexe used and six tes~ ~n~m~
each at the drug dose levels as is shawn in tables 7a and 7b~ Saline was used as the drug vehicle. No drug -toxicity was nobed in the test ~ni~ for either exampl~s lla or llb. No control ~n;~l~ survived beyond day 10 in example lL~ with a mean death date of day 8~3 ~nd ~eyond day ele~en in ~x~rle llb with a mean death date of day 10~1.
For bo~h the control groups and the drug test gro~sg tu~rs were ;n~ur.~ by IP inocula-tion of tumor seed cells on day 2ero follcwed by i~ tiation of drug -trea~ ~ t on day one wherein compolmd 3 was given once daily for 5 days. Test results are expressed as T/C as per example 5.
TABLE 7a Percent Dl'Ug Dose Treated Group Control Group Treated Animals/
mg/Kg Survival Time Survival Time Con-trol Animals 80~ 16.3 ~.3 196%
400 15~2 183%
200 21.5 259%
100 15.5 1~6%
11.3 136%
TABLE 7b Percent Dru~ Dose Treated Group Control Group Treated Animals/
mg/Kg Sur~i~al Time Survival Time Control Animals 6~Q 16.2 10.1 160%
400 15.7 15~%
~00 14.7 145%
100 12.3 121~
13.0 128%
~or Example 12, co~pound 1 was given IP to a ~roup of AKD2F~ r~ce which were i~flicted by intercra~ial inoculation with L~;s ..~, rnah/, ~
~ 7,~
l~mg seed cells to est~ lish brain tu~.ors. The resul-ts in Table 8 bel~w indicate that the IP inoculation of the afflic-ted ani~ls with cQmpound 1 resulted in reduction of the brain tumors indicating succ,essful crossing of the blood brain barrier by compound 1 follawing the IP in~ection lnto the afflic-ted allLmals~
For this -test, 32 control An;mAl~ were used and no control ~n;~ . survived beyond day eleven with a mean death date of da~ 9.6 for the controls~ Eight test ~ni~r~l~ were used for each drug dose le~el with the exception of the 300 mg/Kg level as is shown in table 8 ~elow~ Water wa5 used as the drug vehicle. For both the control ~roup and ~he -test gro~p~ .~3rs were induced on day zeno with i~itiation cf dru~ treatment on day one where compoun~ 1 was given once daily ~or nin2 days. Test results are expressed as T/C as per example 5.
Percent Drug Dose Treated Group ContrQl Group Treated Animalsf mg~Kg Survival Time Survival Time Control Animals 800 21O3 9.6 221%
700 20.3 211%
600 20.3 211%
~0 5 213%
150 20.5 197%
~5 19.0 185%
37.5 16.0 1~6%
25.0 16.6 172%
In a significant number of disease states of the brain of both pa~h~nic and host dysfunction origins, treatment is inhibited b~
the lack of drug transfer across the blood brain barrier. In. certain in-stances wherein appropriate -treatment of a disease state is k2~0wn, com~
plications can arise in treating these diseases when they are located intercra~ially because of the lack of ~ransfer across ~he blocd brain b~rrier of effective concentrations of appropriate che~otheraputic agen~s.
The indication, as seen in table 8, that coI~?~und 1 success~ ~ly crosses t~e blood brain barrier i5 thus very prorrlising for the trea-bment of brain h ~
DJ~
t~nors.
~ he :Eollcwinc3 representat:i,ve e,~ mples, 13 throuyh 17, are gi.ven for the fo~ lations of the active csmpo~md of the inven-ti.on in illustrative pharmaceutical compositions utiliziny illustrative carriers~
In these examples~ ex~nple 13 illustrat:es the use of the compou~lds of the inven-tion in injectables suitable for intra~enous or o~he~ type5 of injec-tion into the host anLmaL. Example 14 is directed to an oral syrup prepar-ation. Example 15 to an oral capsule p.reparation and E~3mple 16 to oral tablets. Exa~ple 17 is di.r.ec-t~d to use of the compoun~s of the invention in suitable suppositories. For Examples 13 through 17 the in~redients a~e listed follcwed by ~he me-thods of preparing the co~çc~ition.
E~MPLE 13 INJECTABLES
Example 13a Csmpound 1 Qo~pound 1 250mg - 1000 mg Water ~or Injectio~ USP q.s.
Cc~npound 1 is dissolved in the water and passecl through a 0.22 ~ filter. Ihe -filtered solution is added to ampoules or vials, sealed and sterilized.
Example 13b Compound 3 Canpound 3 as a Sodium Salt 250 mg - 1000 mg Water for Injection USP q.s.
~repared as per Example 3a, above.
E~AMPLE 14 SYRUP
Example 14a Compound 1 250 mg Active ingredient/5 ml syrup Comp~l~nd 1 sOg Purified Water USP 200 ml Cherry Syr~p ~s. or lOOO ml Cc~npow1d 1 is clissolved in the wa-ter and to ~his solu~ion the syrup is added with mild s-tirring.
Example 14b Compound 3 250 mg Ac-tive ingred:Lerlt/5 ml syrup Compound 3 as a Sodium Salt 50.0 g Purified Water USP q.s. or 200 ml Cherry Syrup q.s. ad 1000 ml Prepared as per Example 14a above.
CAPSULES
Example 15a Compound 1 100 mg, 250 mg or 500 mg Compound 1 50U g Lactose USP, Anhydrous qOs. or 200 g Stero-tex* Powder HM 5 g Combine compound 1 and the Lactose in a t~in-shell blender equipped with an intensifier bar~ Tumble ble~d for two minutes, Eollowed by blending for one minute with ~he intensifier bar and then tumble blend again for one mi~ute. A
port~on of the blend i~s then mixed with the Sterotex* ~owder, passed through a #30 screen and added back to the rema~Lnder o~
the blend. The mixed ingredients are then blended for one minute, blended with -the intensifier bar for thirty seconds and tumble bl.ended for an additional minute Appropri~te si~ed capsules are filled with 141 mg, 352 5 mg or 705 mg of the blend, respectively~ for the 100 mg, 250 mg an~ 500 mg con~
taining capsules.
EXAMPLE 15b Example 15b Compound 2 100 mg, 250 mg or 500 mg Compound 2 *trade marJc 28 -3~
I,actose USP, Anhydrous q.s. or 200 g Sterotex* Powder ~IM 5 ~J
Mix and fill as per Example 15 a.
Example 15c Compound 4 100 mg, 250 mg or 500 mg Compound 4 5~0 g Lactose USP~ ~nhydrous q.sO or 20Q g 5terotex* Powder HM 5 g Mix and fill as per Example 15a EX~MPLE 16 TABLETS
Example 16a Compound 1 100 mgt 200 mg or 500 mg Compound 1 500 g Corn Starch NF 200 g Cellullose, Microcrystalline 46.0 g Sterotex* Powder HM 4.0 g Purified Water q.s. or 300 ml Combine the corn starch, the cellulose and Compoun~
1 -together .in a planetary mixer and mix for two minutes~ Add the water to this combination and mix for one minute. The resulting mix is spread on trays and dried in a hot air oven at 50C until a moisture level of 1 to ? percent is o~tained.
The dried mix is then milled with a Fitzmill through a ~RH2B
screen at medium speed. The Sterotex~ Powder is adde~ to a po~tion o~ the mix and passed through a #30 screen, and added ~ack to the milled mixture and the total blended for five minutes by drum rol.lincJ. Compressed tablets oE 150 m~, 375 mcJ
~ 29 -*trade mark rn;~ 3
7 ~
and 750 mg respec-t:ivel~, of the tota:l mix a:re fo~led with appropriate sized punches for ~.he ~00 mg, 250 mg or 500 mg con-taining tablet~.
EXAMPL:E 17 SUPPOSITORIES
Example 17a Compound 1 250 mg, 500 mg or 1000 mg pex 3 g Compound 1 250 mg500 mg1000 mg Polyethylene Glycol 1925 my1750 mg 1400 mg Polyethylene Glycol 825 mg750 mg 600 mg ~00.0 Melt the Polyethylene Glycol 1540 and the Poly-ethylene Glycol 8000 together at 60C and dissolve Compound 1 into the melt. ~old this total at 25~C into appropriate suppositories.
Example 17b Compound 2 250, 500, 1000 mg pex 3 g Compound 2 250 mg 500 mg 1000 mg Polyethylene Glycol 1925 mg1750 mg 1400 mg 154~
Polyethylerle Glycol ~25 mg750 mg 600 mg ~~
Prepare as per Example 17a a~ove.
mab/~ n
and 750 mg respec-t:ivel~, of the tota:l mix a:re fo~led with appropriate sized punches for ~.he ~00 mg, 250 mg or 500 mg con-taining tablet~.
EXAMPL:E 17 SUPPOSITORIES
Example 17a Compound 1 250 mg, 500 mg or 1000 mg pex 3 g Compound 1 250 mg500 mg1000 mg Polyethylene Glycol 1925 my1750 mg 1400 mg Polyethylene Glycol 825 mg750 mg 600 mg ~00.0 Melt the Polyethylene Glycol 1540 and the Poly-ethylene Glycol 8000 together at 60C and dissolve Compound 1 into the melt. ~old this total at 25~C into appropriate suppositories.
Example 17b Compound 2 250, 500, 1000 mg pex 3 g Compound 2 250 mg 500 mg 1000 mg Polyethylene Glycol 1925 mg1750 mg 1400 mg 154~
Polyethylerle Glycol ~25 mg750 mg 600 mg ~~
Prepare as per Example 17a a~ove.
mab/~ n
Claims (4)
PROPERTY OR PRIVILEGE IS CLAIMED ARE DEFINED AS FOLLOWS:
1 . An antitumor composition for the treatment of malignant tumors in vivo containing as its active ingredient an effective amount of a compound selected from the group consisting of 2-.beta.-D-ribofuranosylthiazole-4-carboxamide, 2-(2,3,5-tri-O-acetyl-.beta.-D-ribofuranosyl)-thiazole-4-carboxamide and physiologically acceptable salts thereof; and a pharmaceutically acceptable carrier for said active ingredient.
2. The composition of claim 1, wherein said compound is 2-.beta.-D-ribofuranosylthiazole-4-carboxamide or a physiologically acceptable salt thereof.
3. The composition of claim 1, wherein said compound is 2 -(2,3,5-tri-O-acetyl-.beta.-D-ribofuranosyl)-thiazole-
4-carboxamide or a physiologically acceptable salt thereof.
Applications Claiming Priority (6)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US21619780A | 1980-12-15 | 1980-12-15 | |
| US216,197 | 1980-12-15 | ||
| US32445581A | 1981-11-24 | 1981-11-24 | |
| US324,455 | 1981-11-24 | ||
| CA000392323A CA1175421A (en) | 1980-12-15 | 1981-12-15 | TREATMENT OF MALIGNANT TUMORS WITH 2-.beta.-D- RIBOFURANOSYLTHIAZOLE-4-CARBOXAMIDE AND RELATED COMPOUNDS |
| CH02572/94A CH688398A5 (en) | 1980-12-15 | 1994-08-22 | 2-beta-D-ribofuranosyl-thiazol-4-carboxamide derivatives |
Related Parent Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA000392323A Division CA1175421A (en) | 1980-12-15 | 1981-12-15 | TREATMENT OF MALIGNANT TUMORS WITH 2-.beta.-D- RIBOFURANOSYLTHIAZOLE-4-CARBOXAMIDE AND RELATED COMPOUNDS |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA1199274A true CA1199274A (en) | 1986-01-14 |
Family
ID=27426338
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA000450522A Expired CA1199274A (en) | 1980-12-15 | 1984-03-26 | TREATMENT OF MALIGNANT TUMORS WITH 2-.beta.-D- RIBOFURANOSYL-THIAZOLE-4-CARBOXAMIDE AND RELATED COMPOUNDS |
Country Status (1)
| Country | Link |
|---|---|
| CA (1) | CA1199274A (en) |
-
1984
- 1984-03-26 CA CA000450522A patent/CA1199274A/en not_active Expired
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