CA2095518A1 - Specific inhibition of dihydrofolate reductase and compounds therefor - Google Patents
Specific inhibition of dihydrofolate reductase and compounds thereforInfo
- Publication number
- CA2095518A1 CA2095518A1 CA002095518A CA2095518A CA2095518A1 CA 2095518 A1 CA2095518 A1 CA 2095518A1 CA 002095518 A CA002095518 A CA 002095518A CA 2095518 A CA2095518 A CA 2095518A CA 2095518 A1 CA2095518 A1 CA 2095518A1
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- Prior art keywords
- diamino
- pyrimidine
- benzyl
- methoxy
- methoxybenzyl
- Prior art date
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- 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
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
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- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Organic Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
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- Communicable Diseases (AREA)
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- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Plural Heterocyclic Compounds (AREA)
Abstract
Compounds derived from pyrimidines having improved activity against fungi such as Pneumocystis carinii, and improved selectivity for P. carinii dihydrofolate reductase over human dihydrofolate reductase, are disclosed. P. carinii pneumonia is advantageously treated with the disclosed compounds.
Description
WO 92/0&16t PCI`/US91/08515 209~
SPECD~C IN~r~oN OF D~DROFOLATE REDUCTAS~
AND COMPOUNDS T~EREFOR
Descri~tion Technical Field This invendon relaus to ph~nracology a~d the inhibi~ion of enzymes specific to pathogens. More pa~icularly, the invendon relates to methods for spe-cifically inhibiting the enzyme dihydrofolate reductase in fungal pathogens, andcompounds therefor.
Back~round of the Invention Pneumocvstis carinii pneumonia (PCP) is a significant, life-threatening infecdon in immunocompromised subjects, and is a leading cause of mosbidity and mor~ality in patients presenting acquired immunodeficiency syndrome (A~S~.
Since the onset of the AIDS epidemic, the incidence of PCP has risen from approximately 200 cases per year to more than 25,000 cases pe~ year in the United States.
Due to the lack of a conunuous in vitro c~ture system, and the cumber-some nature of the rat model of PCP, anti-P. cannii therapy has been developed largely on the assumption that P. carinii was a species of protozoa, and thus that an~-psotozoal agents were likely to be e~Eective. 'rhe two principle thesapeuùc modalities, trimethoprinlsulfame~oxazole and pentamidine, wese developed empirically. Howevèr, P. carinii has recerltly ~een suggested to belong instead to the Kingdom Fungi (J.C. Edman et al., Nature (1988) 334:519-22).
wo 9V08461 pcr/us91/o8sls
SPECD~C IN~r~oN OF D~DROFOLATE REDUCTAS~
AND COMPOUNDS T~EREFOR
Descri~tion Technical Field This invendon relaus to ph~nracology a~d the inhibi~ion of enzymes specific to pathogens. More pa~icularly, the invendon relates to methods for spe-cifically inhibiting the enzyme dihydrofolate reductase in fungal pathogens, andcompounds therefor.
Back~round of the Invention Pneumocvstis carinii pneumonia (PCP) is a significant, life-threatening infecdon in immunocompromised subjects, and is a leading cause of mosbidity and mor~ality in patients presenting acquired immunodeficiency syndrome (A~S~.
Since the onset of the AIDS epidemic, the incidence of PCP has risen from approximately 200 cases per year to more than 25,000 cases pe~ year in the United States.
Due to the lack of a conunuous in vitro c~ture system, and the cumber-some nature of the rat model of PCP, anti-P. cannii therapy has been developed largely on the assumption that P. carinii was a species of protozoa, and thus that an~-psotozoal agents were likely to be e~Eective. 'rhe two principle thesapeuùc modalities, trimethoprinlsulfame~oxazole and pentamidine, wese developed empirically. Howevèr, P. carinii has recerltly ~een suggested to belong instead to the Kingdom Fungi (J.C. Edman et al., Nature (1988) 334:519-22).
wo 9V08461 pcr/us91/o8sls
2 ~ 1 8 Trimethoprim (U.S. Pat. No. 2,909,522) and pyrimethamine, and other dihydrofolate reductase (DHFR) inhibitors are known tO be effective anti-bacterial.
and anti-protozoal agents due to the central role played by DHFR in the metabolic synthesis of nucleic acid precursors. Despite their efficacy when used in con-5 junction with a sulfonarnide, trimethoprim and pyrimetharnine are alone poorinhibitors of P. carinii DHFR. For example, trimethopnm and pyrimethamine exhibit 50% inhibition concentrations (IC50) of 8 and 2,500 nM for E. coli DH~;R, while IC50s for P. car~ii DHFR are 39,600 and 2,4~0 nM, respectively. Other anti-folates have been shown to be more effective inhibitors of P. carinii DHFR,10 but require concornitant administration of leucovorm to prevent toxicity to the host. Allegra et al. (U.S. Pat; No. 4,694,007) suggested treatrnent of P. cannii and Toxoplasmosis gondii with 2,~diarnino-5-methyl-6-[(3,4,5-trimethoxyanilino)-methyl]quinazoline (trimetrexate~, on the theory that the DH~ enzyme in this pathogens is more similar to mammalian DHF~ than to prokaryotic DHF~.
Prior to the AIDS epidemic, these types of agents were sufficient for treatment of the rare cases of P. carinii pneumonia However, in the HIV-positivepatient, therapy and prophylaxis with the standard anti-P. carinii agents are com-plicated by frequent toxic and allergic side effects. New compounds that surpassthe efficacy of the hlown antifolates in treating PCP are desirable. especially 20 inhibitors having greater selectivity for P. carinii DHFR relative to the host (par-ticularly human) DHFR than known inhibitors such as trimethoprim.
Commonly-owned copending U.S. Patent Applicasion Serial No. 447,181, filed 7 December 1989 disclosed several DHFR inhibitors exhibiting good selectivity for P. carinii DHFR.
, Wo92/08461 PCrtUS91/08515 2~9 ,~1~
and anti-protozoal agents due to the central role played by DHFR in the metabolic synthesis of nucleic acid precursors. Despite their efficacy when used in con-5 junction with a sulfonarnide, trimethoprim and pyrimetharnine are alone poorinhibitors of P. carinii DHFR. For example, trimethopnm and pyrimethamine exhibit 50% inhibition concentrations (IC50) of 8 and 2,500 nM for E. coli DH~;R, while IC50s for P. car~ii DHFR are 39,600 and 2,4~0 nM, respectively. Other anti-folates have been shown to be more effective inhibitors of P. carinii DHFR,10 but require concornitant administration of leucovorm to prevent toxicity to the host. Allegra et al. (U.S. Pat; No. 4,694,007) suggested treatrnent of P. cannii and Toxoplasmosis gondii with 2,~diarnino-5-methyl-6-[(3,4,5-trimethoxyanilino)-methyl]quinazoline (trimetrexate~, on the theory that the DH~ enzyme in this pathogens is more similar to mammalian DHF~ than to prokaryotic DHF~.
Prior to the AIDS epidemic, these types of agents were sufficient for treatment of the rare cases of P. carinii pneumonia However, in the HIV-positivepatient, therapy and prophylaxis with the standard anti-P. carinii agents are com-plicated by frequent toxic and allergic side effects. New compounds that surpassthe efficacy of the hlown antifolates in treating PCP are desirable. especially 20 inhibitors having greater selectivity for P. carinii DHFR relative to the host (par-ticularly human) DHFR than known inhibitors such as trimethoprim.
Commonly-owned copending U.S. Patent Applicasion Serial No. 447,181, filed 7 December 1989 disclosed several DHFR inhibitors exhibiting good selectivity for P. carinii DHFR.
, Wo92/08461 PCrtUS91/08515 2~9 ,~1~
- 3 -Disclosure of the Invendon We have now found compounds which exhibit superior activity against D~;R from fungal pathogens. such as P. carinii. and which exhibit much higher selectivitY for the fungal en~me as compared with the mammalian (human) S enzyme. These compounds are represented generally by For~nula I:
Formula I:
R
~ ,q NHq 20 wherein R1 is 3-R3-4 R4-5-R5-benzyl or (N-R6)-8-azabicyclo~3.2.1]oct-3-yl; and ,~\N-R, R~ is H; or R1 and R, together form f~ ~/ where R~ and R5 are LldepeIldently selected from the group consisting of H, lower aL~coxy, lower aLkylthio, lower alkylsulfinyl, vinyl, carboxy-lower aL~cyl, carboxy-lower aL~coxy, 25 dicarboxy-lower alkyl, dicarboxy-lower aLtcoxy, aryl-lower aL~oxy, arylsulfonyl-lower aL"oxy, arylsulfamido-lower alkoxy, and radicals of formula -O(CH2)"-COR8, where n is an integer from 0 to 6 and R~, is an amino acid: R4 isselected from the group consisting Gf lower alkoxy, aryl-lower aL~coxy, lower aLkylthio, halo, lower alkenyl, lower aLIcenyloxy, and pyrrolyl; wi~ the proviso Wo 92~08461 Pcr/us91/n85J5 2Q95~1~
Formula I:
R
~ ,q NHq 20 wherein R1 is 3-R3-4 R4-5-R5-benzyl or (N-R6)-8-azabicyclo~3.2.1]oct-3-yl; and ,~\N-R, R~ is H; or R1 and R, together form f~ ~/ where R~ and R5 are LldepeIldently selected from the group consisting of H, lower aL~coxy, lower aLkylthio, lower alkylsulfinyl, vinyl, carboxy-lower aL~cyl, carboxy-lower aL~coxy, 25 dicarboxy-lower alkyl, dicarboxy-lower aLtcoxy, aryl-lower aL~oxy, arylsulfonyl-lower aL"oxy, arylsulfamido-lower alkoxy, and radicals of formula -O(CH2)"-COR8, where n is an integer from 0 to 6 and R~, is an amino acid: R4 isselected from the group consisting Gf lower alkoxy, aryl-lower aL~coxy, lower aLkylthio, halo, lower alkenyl, lower aLIcenyloxy, and pyrrolyl; wi~ the proviso Wo 92~08461 Pcr/us91/n85J5 2Q95~1~
- 4 -that R3, R~, and R5 are not simultaneously methoxy; R6 is selected from the group consisting of unsubstituud aryl and aryl substituted with one to three radicals selected from the group consisting of halo, lower alkyl, lower aL~oxy, lower alkyl-thio, carboxy. carbamido. carboxy-lower alkyl, and carbamido-lower alkyl; and R75 is selected from the group consisting of aryl and aryl-lower alkyl, where aryl may be substituted with one to three radicals selected from the group consisting of halo, lower alkyl, lower alkoxy, lower allcylthio, carboxy, carbamido, carboxy-lower alkyl, and carbamido-lower aL~cyl; and lower alkyl esters. amides thereof,and pharmaceutically acceptable addition salts.
One aspec~ of the inven~on is a method for treating a fungal infection (such as P. cannii) in a marnmal by administe~ing an effective amount of a compound of foqmula I.
Another aspect of the invendon is a composition for treadng a fungal infection (such as P. carinii) in a mammal compnsing an effecdve amount of a 15 compound of formula I in combinadon with a pha~naceudcally acceptable excipient.
Another aspect of the invendon is the use of a compound of formula I to prepare a composidon for treating a fungal infecdon (such as P. carinii) in a mam-mal comprising an effective amount of a compound of formula I in combinadon 20 with a pharmaceudcally acceptable excipient.
Modes of Carrying Out The Invendon A. Definitions The terms "fungal infecdon" and "fungal pathogen" refer to the infecdon 25 af a mammal with an organism of the Kingdom Fungi, for example Pneumocvsds i, Aspergillus, Candida, Fusanum, and the lilce. The presently preferred method of the invention is the treatment of Pneumocvstis ca~ ii using the compounds of the invendon.
! Wo 9Z/08461 2 ~ 9 S ~ 1 8 Pcr/US91/08515 The term "pharmaceutically acceptable" refers to compounds and compo-sitions which may be administered to mammals without undue toxicity. Exem-plaIy pharmaceutically acceptable salts include mineral acid salts such as hydro-chlorides. hydrobromides, phosphates. sulfates. and the like; and the salts of
One aspec~ of the inven~on is a method for treating a fungal infection (such as P. cannii) in a marnmal by administe~ing an effective amount of a compound of foqmula I.
Another aspect of the invendon is a composition for treadng a fungal infection (such as P. carinii) in a mammal compnsing an effecdve amount of a 15 compound of formula I in combinadon with a pha~naceudcally acceptable excipient.
Another aspect of the invendon is the use of a compound of formula I to prepare a composidon for treating a fungal infecdon (such as P. carinii) in a mam-mal comprising an effective amount of a compound of formula I in combinadon 20 with a pharmaceudcally acceptable excipient.
Modes of Carrying Out The Invendon A. Definitions The terms "fungal infecdon" and "fungal pathogen" refer to the infecdon 25 af a mammal with an organism of the Kingdom Fungi, for example Pneumocvsds i, Aspergillus, Candida, Fusanum, and the lilce. The presently preferred method of the invention is the treatment of Pneumocvstis ca~ ii using the compounds of the invendon.
! Wo 9Z/08461 2 ~ 9 S ~ 1 8 Pcr/US91/08515 The term "pharmaceutically acceptable" refers to compounds and compo-sitions which may be administered to mammals without undue toxicity. Exem-plaIy pharmaceutically acceptable salts include mineral acid salts such as hydro-chlorides. hydrobromides, phosphates. sulfates. and the like; and the salts of
5 organic acids such as acetates, propionates, malonates, benzoates, and the like.
The term "effective amount" refers to an amount of compound sufficient to exhibit a detectable therapeutic effect. The therapeutic effect may include, for example, without limit~tion, inhibiting the growth of pathogens, inhibitmg or pre-venting the release of toxins by pathogens, killing pathogens, and preventing the 10 establishmeM of infection (prophyla~cis). The precise effecdve amount for a sub-ject will depend upon the subject's size and health, the nature of the pathogen, ~e severity of the infection, and the like. Thus, it is not possible to specify an exact effective amount in advance. However, the effective amount for a given situa~ioncan be determined by routine experimentation based on the information provided 15 herein.
The term "lower alkyl" refers to saturated straight or branched-chain rad-icals consisting of carbon and hydrogen having from 1 to 6 carbon atoms, inclu-sive. such as methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl, t-butyl, n-hexyl, and the like. '~ower al~oxy" refers to a radical of the form R-O-, where '~" is 20 lower alkyl as defined above. Suitable examples include methoxy, ethoxy, prop-oxy, butoxy, and the like. Similarly, "lower aL~cylthio" refes to radicals of the form R-S-, and 'qower aLkylsulfinyl" refes to groups of the form R-S(-O)-. For example, one may employ methylthio, ethylthio, methylsulfi~yl, t-butylsulfinyl, and the li~e. 'I,ower alkenyl" refers to stIaight or branched-chain radicals consist-25 ing of carbon and hydrogen having 2-6 carbon atoms and at least one double bond between a pair of carbon atoms, such as ethenyl (vinyl), 2-propenyl, l-methyl-- ethenyl, 2-butenyl, 3-butenyl, and the like.
Wo 92/08461 PCr/US91/08A~l5 2ass~18
The term "effective amount" refers to an amount of compound sufficient to exhibit a detectable therapeutic effect. The therapeutic effect may include, for example, without limit~tion, inhibiting the growth of pathogens, inhibitmg or pre-venting the release of toxins by pathogens, killing pathogens, and preventing the 10 establishmeM of infection (prophyla~cis). The precise effecdve amount for a sub-ject will depend upon the subject's size and health, the nature of the pathogen, ~e severity of the infection, and the like. Thus, it is not possible to specify an exact effective amount in advance. However, the effective amount for a given situa~ioncan be determined by routine experimentation based on the information provided 15 herein.
The term "lower alkyl" refers to saturated straight or branched-chain rad-icals consisting of carbon and hydrogen having from 1 to 6 carbon atoms, inclu-sive. such as methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl, t-butyl, n-hexyl, and the like. '~ower al~oxy" refers to a radical of the form R-O-, where '~" is 20 lower alkyl as defined above. Suitable examples include methoxy, ethoxy, prop-oxy, butoxy, and the like. Similarly, "lower aL~cylthio" refes to radicals of the form R-S-, and 'qower aLkylsulfinyl" refes to groups of the form R-S(-O)-. For example, one may employ methylthio, ethylthio, methylsulfi~yl, t-butylsulfinyl, and the li~e. 'I,ower alkenyl" refers to stIaight or branched-chain radicals consist-25 ing of carbon and hydrogen having 2-6 carbon atoms and at least one double bond between a pair of carbon atoms, such as ethenyl (vinyl), 2-propenyl, l-methyl-- ethenyl, 2-butenyl, 3-butenyl, and the like.
Wo 92/08461 PCr/US91/08A~l5 2ass~18
- 6 -The term "carboxy-lower aL~cyl" refers to radicals havirlg the form -(CH2)n-COOH, where n is an integer from 1 to 6 inclusive. 'l)icarboxy-lower alkyl" indicates lower alkyl chains having two COOH groups attached.
"Aryl" denotes cyclic hydrocarbon radicals of 6-10 carbon atoms which 5 exhibit aromatic character, for example phenyl and napthyl.
The term "halo" refers to fluoro, chloro, bromo, and iodo. The term "amino acid" refers to any of the 20 or so cornmonly occmring amino acids, for exarnple, glycine. alanine, arginine, phenylalanine, glutamic acid, vali~e, histidine, prolinè. ornilhine, norleucine. and the lilce. When attached as Ro in a radical of 10 the form -O(CH2)n-COR~, the amino acid will preferably be attached via a pepude bond l e. by a bond between the amino group of the amino acid and tne acyl car-bon of the radical.
Tne term "coadministering" means ad~ninistration of a compound of the invention in combination with a second therapeutic agent. The second therapeutic15 agent is a dihydropteroau synthase inhibitor, preferab}y dapsone or a sulfa drug.
Suitable sulfa drugs include, without lLtnitation, sulfadiazine, sulfamethoxazole, and the like. Coadm~Iustration may be simultaneous, for example by administer-ing a mixture of the therapeutic agents, or may be accomplished by adminis~ationof the agents separately within a short time period.
B. General Method The compounds of the invendon are struct3Ially relaud to the compound tnmelhoprim (2,4 diamislo-5-(3,4,5-trimetho~cybenzyl)pylinudine), the synthesis for which is known ifl the att. See for e~ample U.S. Pat. No, 2,909,522, which des-25 cribes the synthesis of tTimethop~im and related compounds. Compounds of for-mula I may similarly be synthesized by those of ordinaIy sldll ifl the art.
Syntheses of such compounds are described in the following U.S. patents:
Hitchings et aL (2,658,897); Hitchings et al. (2,909,522); Hoffer (3,341,541);
Roth (3,772,289); Roth et al. (3,819,629); Roth (3,822,264); Kompis et aL
.
\
~o 92~08461 PCr/US~1/08515 . 2095:)18 (3,931,181); Herrling (3,980,649); Liebenow et al. (3,992,379); Kompis (4,024,145); Rosen (4,033,962); Kompis et al. (4,039,543); Jernow et al.
(4,075,209); Perun et al. (4,087,528); Fritschi et al. ~4,180,578); Kompis et al.
(4,203.980); Poe et aL (4,258,045); Daluge et al. (4,438,267); Dall'Asta (4,485,248); Kompis et al. (4,515,948); Swaringen et al. (4,568,744); Roth et al.
(4,587,341); Kompis et aL (4,590,270); Daluge et al. (4,603,136); Kompis et al.
(4,792,557); and Seydel et aL (4,912,112).
Presently prefelTed compounds of the invention are:
2.4-diamin~5-[3,5-dimethoxy-4 (2-hydro~yprop-2-yl)benzyl]pyr~nidine;
2,~diamino-5-(3,5-dimetho~y-4-N-pyIrolylbenzyl)pyIimidine;
2,4-diamino-5-(3,5 -diethoxy-4-N-pyrrolylbenzyl)pyrimidine;
2,4-diamino-5-(3,5-dipropoxy-4-N-pyrrolylbenzyl)pyrimidine;
2,4-diamino-5-(3,5-dibutoxy-~N-pyrrolylbenzyl)pyrimidine;
2,4-diamino-5-(3,5-diethoxy-4-carboetho~ybenzyl)py~imidine;
2,4-diamino-5-(3,5-divinyl-4-vinyloxybenzyl)pyrimidine;
2,4-diamino-5-[3-(~N-acetaminophenyl)sulfonaminoethoxy-4,5 -dimethoxybenzyl]-pyrimidine;
2.4-diam~no-5-[3-(~aminophenyl)sulfonaminoethoxy-4-methoxybenzyl]pyrim~dine;
2,4-diamino 5-[3-(~N-acetaminophenyl)sulfonaminoetho~y-4 bromo-5-methoxybenzyl]-pyIimidine;
2,4-diamino-5-[3-(~aminophenyl)sulfonaminoethoxy-4 bromo-5-methoxybenzyl]-pyrimidine;
2,4 diamin~5-(3-methoxy-4,5-dibenzyloxybenzyl)pyr~midine;
2,4 diamino-5-(4 benzyloxybenzyl)pynmidine;
2,~diam~no-5-(3,4-dibenzylo~y-5-metho~cybenzyl)pyIimidine;
2,~diamino-5-(3,4 dimethoxy-5-benzylo~cybenzyl)pynmidine;
2,~diamino-5-[3,5-diethoxy-4-(propen-2-yl)benzyl]pyrimidine;
2,4-diamino-5-[3,5-dimethoxy-4-(propen-2-yl)benzyl]pynmidine;
2,~diamino-5-(3,5 -dimethoxy-4-methylthiobenzyl)pyrimidine;
Wo 92/08461 Pcr/US91/08~l5 2~9~j~'i8 2,~diamino-5-(3-methylsulfinyl-4-methoxy-S-methylthiobenzyl)pynmidine;
2,4-dian~no-5-[3-(4,6-dicarboxyhexyloxy) 1 bromo-5-methoxybenzyl]py~imidine;
2,~diamino-5-[3-(3-carboxymethylamino-3-oxopropoxy)-4-methoxybenzyl]-pyrimidine;
5 2,4-diamino-5-(3-[3-~1,3-dicarboxypropyl)amin~3-oxopropoxy]-4-bromo-5-methoxy-benzyl)pynmidine;
2,4-diam~no-5-[3 ,5-dimethoxy-4-((2-phenylsulfinyl)acetyl)benzyl]pyrill~idine;
2,~diamino-5-~3-amino-4-methyl-5-(N-pyrrolyl)benzyl]pyrimidine;
- 2.4-diamino-5-(3,5-di-N-pyrrolyl-4-methoxybenzyl)pynmidine;
102~diamino-5-[3,$-di-me~hoxy-4-(3-hydrocarboxy-l-oxopropylamino)benzyl]pyrimidine;
2,4-diamino-5-[3,5-dimethoxy-(4-acetaminophenylsulfonamino)benzyl]pyrin~idine;
2,4-diamino-5-(3 ,5 -dimethoxy-4-propylbenzyl)pyrimidine;
2,4-diamino-5-(3,5-dic'nloro-4-N-pyrrolylbenzyl)pyrimidine;
2,4-diamino-5-[3,5-dimetnoxy-~1 (2-(2-(2-metnoxy)ethoxy)ethoxy)ethoxybenzyl]-lS pynmidine;
2,4-diamino-S- [3 -(3-benzyloxycarbonylrnethylamino-3-oxopropoxy)-4-bromo-S -methoxy-benzyl]pyrimidine;
2,4-diamino-5-[3-(3-carboxymethylamino-3-oxopropoxy)-4 bromo-S-methoxybenzyll-pyrimidine;02.4-diamino-5-[3-methoxy-~bromo-5-(4 methylaminobenzamidoethoxy)benzyl]-pyrimidine;
3-(2,4-diaminopyrimidin-S-ylmetnyl)-8-(3 ,5-dimethoxyphenyl)-8-azabicyclo[3.2.1]octane;
2H,3H-dihydro-S -(2,4-diamlnopyrimidin-S-ylme~hyl)-6,7-dimethoxybenzofuran;
25 5-(2,~diaminopyrimi~in-5-yknethyl)-7-methoxy-8-bromo-1,2-benzopyran;
5-(2,4~ninopyrimidin-5-ylmethyl)-7,8-dimethoxy-1,2-benzopyran;
2,4-diamino-5-[3-phenyl-5-(3-metho~cy~opoxy)benzyl]pyrimidine;
2.4-diamino-7-(3 ,5-dimethoxybenzyl)pyrrolo[2,3-flql~inazoline;
2.4-diarnino-S - [6-(4-methoxybutoxy)naphth- 1 -yl]pyrimidine;
- ~O 92/08461 2 0 3 ~ Pcr/US91/085tS
2,~diamino-5-(2,7-dimethylbenzpyrazol-5-ylmethyl)pynmidine;
2.4-diamino-5-(4,5,6-trimethoxy-2,3-dihydroinden-l-yl)pyrimidine; and 2,2 -dimethyl-5 -(2,4-diaminopyrimidin-5 -ylmethyl)-7-methoxybenz[b]dioxolane .
The most preferred compounds at present are 2~4 diamino-5-(3,5-diethoxy-4 N-pyrrolylbenzyl)pyrimidine and 2,4 diam~no-j-[3,5-dimethoxy-4-(2-hydroxyprop-2-yl)benzyl]pyrimidine.
Compositions of the invention for administration will generally include an effecuve amount of a compound of formula I in addition to a pharmaceutically acceptable excipient. Suitable excipients include most carriers approved for oral or parenteal administration. including water. saline. Ringer's solution. Hank's solution, and solutions of glucose, lactose, dextrose, ethanol, glyceroL albun~in, and the like. These compositions may optionally include stabili7ers~ antioxidants, antimicrobials. preservatives, buffering agents, surfactants, and other accessory additives. A presently preferred vehicle comprises about l mglmL serum alburrun in phosphate-buffered saline (PBS). A thorough discussion of suitable vehicles for parenteral administration may be found in E.W. Martin, '~emmgton's Pharrna-ceutical Sciences" (Mack Pub. Co., current edition).
The precise dosage necessary will vary with the age, size. and condition of the subject, the nature and severity of the disorder to be treated. and the like:
thus, a precise effective amount carmot be specified in advance. However, appro-priau amounts may be determined by routine experimentation with animal models.
general terms, an effective dose of compound of formula I wi~l range from about lO ,ug/Kg to about 50 mg/Kg. Suitable animal models include the mouse model illustrated in the Examples below. Rats and other rodents have DHFR very similar to the human enzyme, and thus make suitable anirnal models.
A group of exper~nental animals is inoculated with lO-lO0 LD~os of Pneumocvstis ~i, followed by treatment with a soludon of test compound. A negative con-trol group is left untreated, while a positive control group is treated with a stan-dard therapy, such as trimethoprim. Administration of t~le compounds is prefer-wo 92/08461 Pcr/ussl/o8sls 209aal8 ably per os ~, using a gavage), but may be parenteral, for example by subcu-taneous or intramuscular injection, or by inhalation of an aerosol. The animals are monitored during treatment. and are sacrificed and e~amined after 60 days for presence of infection.
C. Exam~les The examples presented below are provided as a further guide to the pracdtioner of ordinary skill in the a~, and are not to be construed as limiting the invention in any way.
E~amPle 1 (Demonstration of Activity) A. Materials Buffers were prepared as follows:
4xDHFR buffer: 200 mM Tes, 300 mM BME, 4 mM EDTA, pH 7Ø
+DHF buffer. 2.5 mg/mL BSA, 0.25 mM NADPH, 625 tlM dihydrofolate. 2.5x DHFR
buffer.
-DHF buffer: 2.5 mglmL BSA, 0.25 mM NADPH, 2.5x DH~;R buffer.
Enzvme buffer 50 mM Tes. S mM DlT, 1 mM EDTA, 20% glycerol, 1 mg/mL BSA, pH 7Ø
Dilution buffer. 50 mM Tes, S mM DTT, 1 mM E~DTA, 1 mg/mL BSA, pH 7Ø
PcDHFR: S llg/mL P. carinii DHFR in enzyme buffer.
crude hDHFR: crude recombinant human DHFR (obtained ~om Hoffmann-LaRoche) in enzyme buffer (9.9 mglmL total protein).
25purifled hDHFR: purified recombinant human DH~;R (obtained from Hoffmann-LaRoche) in enzyme buffer (3.5 mg/mL total protein).
Test compounds were prepared and provided by Ho~mann-LaRoche.
Stock solutio~s were prepared by dissol~ing 2-8 mg in dimethylsulfoxide (DMSO) . .
2,~95'~
to prepare 50 mM solutions. Compounds which did not dissolve at 50 mM were diluted serially to 25, 16.6, or 12.5 mM.
B. Assav Eight compounds (6.7 ,uL each) were added to wells in one column of a S Falcon~ 96-well micro~iter plate, and diluted with 160 llL of water. The remairling wells received 126 ~IL of water. The solutions in the f~st column were then serially diluted (4x) into columns 2-9, with colurnn 10 containing a control sample, and columns 11-12 containing water blanks. The final compound concentrations were 1 mM to 15 nM (after the rema~ng reagents were added).
Using an 8~hannel pipet, 100 ,uL of -DHF buffer were added to each well in columns 11 and 12 (blanks containing only water). Next. 100 ~IL of +DHF buffer were added to each well in colurnns 1-10. PcDHFR or crude or pu~ified rhDHFR enzyme was then added (25 pL) to each well, and the plate contents mLxed using a Titertek shaker. The plates were read on a Molecular Devices Plate Reader at an absorbance of 340 nm, -0.05 O.D. scale. for 10 min-utes reading every 10 seconds. The kinedc data was analyzed using Delta Soft software (Biometallics), and the IC50 and Ki calculated for both human and P.
carinii enzymes. Selectivity was calculated as Ki~n"",~"/Ki,E ~""",; thus, higher values for selectivity indicate that the compound inhibits the P. carinii enzyme to 20 a greater degree than the human enzyme. The results are shown in Table 1 below.
For purposes of comparison, trimethoprim in this assay e~hibits an IC50 of 20 with a selectivity of 0.1.
wo 92/08461 pcr/us91/o8~l$
2~ -8 TABLE l: Compounds of Fonnula I
R3 R4 R5 IC50 Selectivity S
-OMe -OMe -O(CH2)2NHSO2~NHAc 2. ~M 350 -OMe -CMe2OH -OMe 25 ~M 350 10-OEt pyrrolyl -OEt 2.4 ~M 310 -OMe pyrrolyl -OMe 29 ~M 30 vinyl vi~yloxy vinyl 3.4 ~M 250 H BzO- H 16 ~M 50 -OMe BzO- BzO- 8.3 ~M 100 20-OMe -C(Me)=CH2 -OMe 24 ~M 160 -OEt -C(Me)=CH2 -OEt 2.5 ~M 160 -OMe -SMe -OMe 19 ~M 145 -SMe -ONe -S(O)Me 18 ~M 490 -OMeBr -O(CH2)2C~(CH2~2COOHO.04 ~M 525 COOH
-OMeBr -O(CH2)2CO(N-Asp) 2 ~M 4300 H-OMe -O(CH2)2CO(N-Gly) 9.3 ~M 80 -OMeBr -O(CH2)2CO(N-Gly) 10 ~M 8 Me -CH3, E~ - -CH2CH3, Bz - -CH2C6k6, N-~ly - -NHCH2COOH, N-Asp --NHCH(COOH)CH2COOH, ~- phenyl, Ac - -COCH3 Example 2 Proceeding as described in E~ample l above, the compounds listed below were assayed and found to exhibit high activity and selectivity, as set for~ in Table II:
A) 2,~diamino-5-(~benzylo~cybenzyl)pyTimidine;
Wo 92/084~l PCr/US91/08515 " 209~.jl8 B) 2,~dia~r.uno-5-(3,~dimethoxy-5-benzyloxybenzyl)pyrimidine:
C) 2,4 diarr.uno-5-(3,4 dibenzyloxy-5-methoxybenzyl)pyrirr.udine;
D) 2,~diamino-5-[3,5-dimethoxy-~(2-hydroxyprop-2-yl)benzyl~pyrilI~dine;
E) 2,~diamino-5-(3,5-dimethoxy- 1 N-pyrrolylbenzyl)pynmidine:
F) 2,1 diammo-5-(3,5-diethoxy-4-N-pyrrolylbenzyl)pyIimidine;
G) 2,1 diamino-5-(3~5-divinyl-4 vinyloxybenzyl)pyrimidine;
H) 2,4-diamino-5-[3-(l N-acetaminophenyl)sulfonaminoethoxy-4,5 dimeth-oxybenzyl]pyrim~dine;
I) 2,~dia~runo-5-[3-(4 aminophenyl)sulfonatninoethoxy-~methoxybenzyl]-pyri~udine;
J) 2,~diamino-5-[3-(4N-acetaminophenyl)sulfona n~noethoxy-~l bromo-5-methoxybenzyl]pyrimidine;
K) 2,1 diarnino-5-[3-(~aminophenyl~sulfonaminoethoxy-~bromo-5-methoxy-benzyl]pyrimidine;
L) 2,~diamino-5-[3,5-diethoxy-1 (propen-2-yl)benzyl]pyr~idine;
M) 2,~1 diamino-5-[3,5-dimethoxy-1 (propen-2-yl)benzyl]pyrimidine;
N~ 2,1 diarnino~5-(3,5-dimethoxy-~methylthiobenzyl)pyrimidine;
O) 2,~diamino-5-(3-methylsulfinyl-~methoxy-5-methylthiobenzyl)-pyrirnidine;
P) 2,4-diamino-5-[3-(4,6-dicarboxyhexyloxy)-4-bromo-5-methoxybenzyl]-pyrimidine;
Q) 2,4-diamino-5-[3-(3-carboxymethylamino-3-oxopropoxy)-l methoxy-benzyl]pynmidine;
R) 2,~diamino-5-(3-[3-(1,3-dicarboxypropyl)amino-3-oxopropoxy]-1 bromo-5-methoxybenzyl)pylimidi~e;
S) 2,~dian~no-5-[3,5-dimetho~cy-~((2-phenylsulfonyl)acetyl)benzyl]-pyrimidine;
T) 2,~diam~no-5-[3-am~no-4 methyl-5-(N-pyIrolyl)benzyl]pynmidine;
U) 2,4 diamino-5-(3,5-di-N-pyrrQlyl-4 methoxybenzyl)pyrimidine;
Wo 92/08461 PCr/US91/08~.15 2~9~18 V) 2,4-diamino-5-[3,5-di-methoxy-4 (3-hydrocarboxy-1-oxoprowlamino)-benzyl]pyri~udine;
W) 2,4-dian~ino-5-[3,5-dimethoxy-(~acetaminophenylsulfonarnino)benzyll-pyr~midine;
5 X) 2,~diamino-5-(3,5-dimethoxy-~propylbenzyl)pyrimidine;
Y) 2,4-diarnino-5-(3,5-dichloro-4-N-pyrrolylbenzyl)pyrimidine;
Z) 2,~diamino-5-[3,5-dimethoxy-4 (2-(2-(2-methoxy)ethoxy)ethoxy)ethoxy-benzyl]pyrimidine:
AA) 2,4-diarnino-5-[3-(3-benzyloxycarbonylmethylamino-3-oxopropoxy)-~
bromo-S-methoxybenzyl]pyrimidine;
BB) 2,4-diamino-5-[3-(3-carboxymethylamino-3-oxopropoxy)-4-bromo-5-methoxybenzyl]pyri~udine;
CC) 2,4-diamino-5-[3-methoxy-4-bromo-5-(4-methylaminobenzamidoethoxy)-benzyl]pyrimidine;
DD) 3-(2,1 diaminopyrimidin-5-ylmethyl)-8-(3,5-dimethoxyphenyl)-8-aza-bicyclo[3 .2.1 ]octane;
EE~ 2~I,3H-dihydro-5-(2,~diaminopy~imidin-5-ylmethyl)-6,7-dimethoxybenzo-furan;
FF) 5-(2,4-diaminopyrimidin-5-ylmethyl)-7-methoxy-8-bromo-1.2-benzopyran;
GG) 5-(2,~diaminopyrimidin-5-ylmethyl3-7,8-dimethoxy- 1 ,2-benzopyran;
HH) 2,~diamino-5-[3-phenyl-5-(3-methoxypropoxy)benzyl]pyr~Irudine:
II) 2,'1 diasnino-7-(3,5-dirnethoxybenzyl)pyrrolo[2,3-f~quinazoline;
JJ) 2,1 diamino-5-[~(4 methoxybutoxy)naphth-1-yl]pyrimidine;
KK) 2,4 diamin~5-(4,5,~tnrnethoxy-2,3 dihydroinden-1-yl)pynmidine;
LL) 2,2-dimethyl-5-(2,~diaminopyriIrLidin-S-ylmethyl)-7-methoxybenz[b]di-oxolane;
MM) 2,~diamino-5-(3,5-diethoxy-1 carboethoxybenzyl)pyrimidine; and NN) 2,1 diamino-5-(2,7-dimethylbenzpyrazol-5-yLmethyl)pyrirrudine.
WO 92/0&~1 PCT/US91/08515 2~3;~ 1;8 TABLE ~:
¦ Compound IC50 Selectivity ¦
_ _, A) 16.0 53.2 B) 13.9 3.3 C) 8.3 90.3 ¦ D) 24.8 343.5 E) 2.9 29.4 F) 2.4 312.4 G) 3.4 250.5 H) 2.4 248.5 I) 0.57 7.8 J) 3.7 29.5 ~) 0.97 12.1 L) 2.5 190.9 M) 24.0 159.8 N) lg.o 143.5 O) 17.5 486.8 P) 0.043 528.0 Q) 9.3 77.7 R) 2.0 4255.0 S) 14.8 36.8 T) 33.5 42.5 U) 4.5 47.7 V) 100.0 85.2 W) 31.0 14.3 X) 20.0 42.6 Y) 10.9 53.2 Z) 36.0 33.1 WO92~08461 PCT/US91/08,C.15 ~9 -~8 TABLE II (continued):
Compound IC50Selectivity AA) 7.9 14.4 BB) 10.0 13.1 CC) 7.8 14.6 DD) 0.016 31.0 EE) 12.9 26.4 FF) 5.9 6.9 GG) 11.5 266.7 HH) 12.2 11.2 II) 0.039 1.2 JJ) 80.0 106.5 KK) 87.0 97.9 I LL) 13.9 15.0 NM) 8.3 63.6 I NN) 18.0 10.7
"Aryl" denotes cyclic hydrocarbon radicals of 6-10 carbon atoms which 5 exhibit aromatic character, for example phenyl and napthyl.
The term "halo" refers to fluoro, chloro, bromo, and iodo. The term "amino acid" refers to any of the 20 or so cornmonly occmring amino acids, for exarnple, glycine. alanine, arginine, phenylalanine, glutamic acid, vali~e, histidine, prolinè. ornilhine, norleucine. and the lilce. When attached as Ro in a radical of 10 the form -O(CH2)n-COR~, the amino acid will preferably be attached via a pepude bond l e. by a bond between the amino group of the amino acid and tne acyl car-bon of the radical.
Tne term "coadministering" means ad~ninistration of a compound of the invention in combination with a second therapeutic agent. The second therapeutic15 agent is a dihydropteroau synthase inhibitor, preferab}y dapsone or a sulfa drug.
Suitable sulfa drugs include, without lLtnitation, sulfadiazine, sulfamethoxazole, and the like. Coadm~Iustration may be simultaneous, for example by administer-ing a mixture of the therapeutic agents, or may be accomplished by adminis~ationof the agents separately within a short time period.
B. General Method The compounds of the invendon are struct3Ially relaud to the compound tnmelhoprim (2,4 diamislo-5-(3,4,5-trimetho~cybenzyl)pylinudine), the synthesis for which is known ifl the att. See for e~ample U.S. Pat. No, 2,909,522, which des-25 cribes the synthesis of tTimethop~im and related compounds. Compounds of for-mula I may similarly be synthesized by those of ordinaIy sldll ifl the art.
Syntheses of such compounds are described in the following U.S. patents:
Hitchings et aL (2,658,897); Hitchings et al. (2,909,522); Hoffer (3,341,541);
Roth (3,772,289); Roth et al. (3,819,629); Roth (3,822,264); Kompis et aL
.
\
~o 92~08461 PCr/US~1/08515 . 2095:)18 (3,931,181); Herrling (3,980,649); Liebenow et al. (3,992,379); Kompis (4,024,145); Rosen (4,033,962); Kompis et al. (4,039,543); Jernow et al.
(4,075,209); Perun et al. (4,087,528); Fritschi et al. ~4,180,578); Kompis et al.
(4,203.980); Poe et aL (4,258,045); Daluge et al. (4,438,267); Dall'Asta (4,485,248); Kompis et al. (4,515,948); Swaringen et al. (4,568,744); Roth et al.
(4,587,341); Kompis et aL (4,590,270); Daluge et al. (4,603,136); Kompis et al.
(4,792,557); and Seydel et aL (4,912,112).
Presently prefelTed compounds of the invention are:
2.4-diamin~5-[3,5-dimethoxy-4 (2-hydro~yprop-2-yl)benzyl]pyr~nidine;
2,~diamino-5-(3,5-dimetho~y-4-N-pyIrolylbenzyl)pyIimidine;
2,4-diamino-5-(3,5 -diethoxy-4-N-pyrrolylbenzyl)pyrimidine;
2,4-diamino-5-(3,5-dipropoxy-4-N-pyrrolylbenzyl)pyrimidine;
2,4-diamino-5-(3,5-dibutoxy-~N-pyrrolylbenzyl)pyrimidine;
2,4-diamino-5-(3,5-diethoxy-4-carboetho~ybenzyl)py~imidine;
2,4-diamino-5-(3,5-divinyl-4-vinyloxybenzyl)pyrimidine;
2,4-diamino-5-[3-(~N-acetaminophenyl)sulfonaminoethoxy-4,5 -dimethoxybenzyl]-pyrimidine;
2.4-diam~no-5-[3-(~aminophenyl)sulfonaminoethoxy-4-methoxybenzyl]pyrim~dine;
2,4-diamino 5-[3-(~N-acetaminophenyl)sulfonaminoetho~y-4 bromo-5-methoxybenzyl]-pyIimidine;
2,4-diamino-5-[3-(~aminophenyl)sulfonaminoethoxy-4 bromo-5-methoxybenzyl]-pyrimidine;
2,4 diamin~5-(3-methoxy-4,5-dibenzyloxybenzyl)pyr~midine;
2,4 diamino-5-(4 benzyloxybenzyl)pynmidine;
2,~diam~no-5-(3,4-dibenzylo~y-5-metho~cybenzyl)pyIimidine;
2,~diamino-5-(3,4 dimethoxy-5-benzylo~cybenzyl)pynmidine;
2,~diamino-5-[3,5-diethoxy-4-(propen-2-yl)benzyl]pyrimidine;
2,4-diamino-5-[3,5-dimethoxy-4-(propen-2-yl)benzyl]pynmidine;
2,~diamino-5-(3,5 -dimethoxy-4-methylthiobenzyl)pyrimidine;
Wo 92/08461 Pcr/US91/08~l5 2~9~j~'i8 2,~diamino-5-(3-methylsulfinyl-4-methoxy-S-methylthiobenzyl)pynmidine;
2,4-dian~no-5-[3-(4,6-dicarboxyhexyloxy) 1 bromo-5-methoxybenzyl]py~imidine;
2,~diamino-5-[3-(3-carboxymethylamino-3-oxopropoxy)-4-methoxybenzyl]-pyrimidine;
5 2,4-diamino-5-(3-[3-~1,3-dicarboxypropyl)amin~3-oxopropoxy]-4-bromo-5-methoxy-benzyl)pynmidine;
2,4-diam~no-5-[3 ,5-dimethoxy-4-((2-phenylsulfinyl)acetyl)benzyl]pyrill~idine;
2,~diamino-5-~3-amino-4-methyl-5-(N-pyrrolyl)benzyl]pyrimidine;
- 2.4-diamino-5-(3,5-di-N-pyrrolyl-4-methoxybenzyl)pynmidine;
102~diamino-5-[3,$-di-me~hoxy-4-(3-hydrocarboxy-l-oxopropylamino)benzyl]pyrimidine;
2,4-diamino-5-[3,5-dimethoxy-(4-acetaminophenylsulfonamino)benzyl]pyrin~idine;
2,4-diamino-5-(3 ,5 -dimethoxy-4-propylbenzyl)pyrimidine;
2,4-diamino-5-(3,5-dic'nloro-4-N-pyrrolylbenzyl)pyrimidine;
2,4-diamino-5-[3,5-dimetnoxy-~1 (2-(2-(2-metnoxy)ethoxy)ethoxy)ethoxybenzyl]-lS pynmidine;
2,4-diamino-S- [3 -(3-benzyloxycarbonylrnethylamino-3-oxopropoxy)-4-bromo-S -methoxy-benzyl]pyrimidine;
2,4-diamino-5-[3-(3-carboxymethylamino-3-oxopropoxy)-4 bromo-S-methoxybenzyll-pyrimidine;02.4-diamino-5-[3-methoxy-~bromo-5-(4 methylaminobenzamidoethoxy)benzyl]-pyrimidine;
3-(2,4-diaminopyrimidin-S-ylmetnyl)-8-(3 ,5-dimethoxyphenyl)-8-azabicyclo[3.2.1]octane;
2H,3H-dihydro-S -(2,4-diamlnopyrimidin-S-ylme~hyl)-6,7-dimethoxybenzofuran;
25 5-(2,~diaminopyrimi~in-5-yknethyl)-7-methoxy-8-bromo-1,2-benzopyran;
5-(2,4~ninopyrimidin-5-ylmethyl)-7,8-dimethoxy-1,2-benzopyran;
2,4-diamino-5-[3-phenyl-5-(3-metho~cy~opoxy)benzyl]pyrimidine;
2.4-diamino-7-(3 ,5-dimethoxybenzyl)pyrrolo[2,3-flql~inazoline;
2.4-diarnino-S - [6-(4-methoxybutoxy)naphth- 1 -yl]pyrimidine;
- ~O 92/08461 2 0 3 ~ Pcr/US91/085tS
2,~diamino-5-(2,7-dimethylbenzpyrazol-5-ylmethyl)pynmidine;
2.4-diamino-5-(4,5,6-trimethoxy-2,3-dihydroinden-l-yl)pyrimidine; and 2,2 -dimethyl-5 -(2,4-diaminopyrimidin-5 -ylmethyl)-7-methoxybenz[b]dioxolane .
The most preferred compounds at present are 2~4 diamino-5-(3,5-diethoxy-4 N-pyrrolylbenzyl)pyrimidine and 2,4 diam~no-j-[3,5-dimethoxy-4-(2-hydroxyprop-2-yl)benzyl]pyrimidine.
Compositions of the invention for administration will generally include an effecuve amount of a compound of formula I in addition to a pharmaceutically acceptable excipient. Suitable excipients include most carriers approved for oral or parenteal administration. including water. saline. Ringer's solution. Hank's solution, and solutions of glucose, lactose, dextrose, ethanol, glyceroL albun~in, and the like. These compositions may optionally include stabili7ers~ antioxidants, antimicrobials. preservatives, buffering agents, surfactants, and other accessory additives. A presently preferred vehicle comprises about l mglmL serum alburrun in phosphate-buffered saline (PBS). A thorough discussion of suitable vehicles for parenteral administration may be found in E.W. Martin, '~emmgton's Pharrna-ceutical Sciences" (Mack Pub. Co., current edition).
The precise dosage necessary will vary with the age, size. and condition of the subject, the nature and severity of the disorder to be treated. and the like:
thus, a precise effective amount carmot be specified in advance. However, appro-priau amounts may be determined by routine experimentation with animal models.
general terms, an effective dose of compound of formula I wi~l range from about lO ,ug/Kg to about 50 mg/Kg. Suitable animal models include the mouse model illustrated in the Examples below. Rats and other rodents have DHFR very similar to the human enzyme, and thus make suitable anirnal models.
A group of exper~nental animals is inoculated with lO-lO0 LD~os of Pneumocvstis ~i, followed by treatment with a soludon of test compound. A negative con-trol group is left untreated, while a positive control group is treated with a stan-dard therapy, such as trimethoprim. Administration of t~le compounds is prefer-wo 92/08461 Pcr/ussl/o8sls 209aal8 ably per os ~, using a gavage), but may be parenteral, for example by subcu-taneous or intramuscular injection, or by inhalation of an aerosol. The animals are monitored during treatment. and are sacrificed and e~amined after 60 days for presence of infection.
C. Exam~les The examples presented below are provided as a further guide to the pracdtioner of ordinary skill in the a~, and are not to be construed as limiting the invention in any way.
E~amPle 1 (Demonstration of Activity) A. Materials Buffers were prepared as follows:
4xDHFR buffer: 200 mM Tes, 300 mM BME, 4 mM EDTA, pH 7Ø
+DHF buffer. 2.5 mg/mL BSA, 0.25 mM NADPH, 625 tlM dihydrofolate. 2.5x DHFR
buffer.
-DHF buffer: 2.5 mglmL BSA, 0.25 mM NADPH, 2.5x DH~;R buffer.
Enzvme buffer 50 mM Tes. S mM DlT, 1 mM EDTA, 20% glycerol, 1 mg/mL BSA, pH 7Ø
Dilution buffer. 50 mM Tes, S mM DTT, 1 mM E~DTA, 1 mg/mL BSA, pH 7Ø
PcDHFR: S llg/mL P. carinii DHFR in enzyme buffer.
crude hDHFR: crude recombinant human DHFR (obtained ~om Hoffmann-LaRoche) in enzyme buffer (9.9 mglmL total protein).
25purifled hDHFR: purified recombinant human DH~;R (obtained from Hoffmann-LaRoche) in enzyme buffer (3.5 mg/mL total protein).
Test compounds were prepared and provided by Ho~mann-LaRoche.
Stock solutio~s were prepared by dissol~ing 2-8 mg in dimethylsulfoxide (DMSO) . .
2,~95'~
to prepare 50 mM solutions. Compounds which did not dissolve at 50 mM were diluted serially to 25, 16.6, or 12.5 mM.
B. Assav Eight compounds (6.7 ,uL each) were added to wells in one column of a S Falcon~ 96-well micro~iter plate, and diluted with 160 llL of water. The remairling wells received 126 ~IL of water. The solutions in the f~st column were then serially diluted (4x) into columns 2-9, with colurnn 10 containing a control sample, and columns 11-12 containing water blanks. The final compound concentrations were 1 mM to 15 nM (after the rema~ng reagents were added).
Using an 8~hannel pipet, 100 ,uL of -DHF buffer were added to each well in columns 11 and 12 (blanks containing only water). Next. 100 ~IL of +DHF buffer were added to each well in colurnns 1-10. PcDHFR or crude or pu~ified rhDHFR enzyme was then added (25 pL) to each well, and the plate contents mLxed using a Titertek shaker. The plates were read on a Molecular Devices Plate Reader at an absorbance of 340 nm, -0.05 O.D. scale. for 10 min-utes reading every 10 seconds. The kinedc data was analyzed using Delta Soft software (Biometallics), and the IC50 and Ki calculated for both human and P.
carinii enzymes. Selectivity was calculated as Ki~n"",~"/Ki,E ~""",; thus, higher values for selectivity indicate that the compound inhibits the P. carinii enzyme to 20 a greater degree than the human enzyme. The results are shown in Table 1 below.
For purposes of comparison, trimethoprim in this assay e~hibits an IC50 of 20 with a selectivity of 0.1.
wo 92/08461 pcr/us91/o8~l$
2~ -8 TABLE l: Compounds of Fonnula I
R3 R4 R5 IC50 Selectivity S
-OMe -OMe -O(CH2)2NHSO2~NHAc 2. ~M 350 -OMe -CMe2OH -OMe 25 ~M 350 10-OEt pyrrolyl -OEt 2.4 ~M 310 -OMe pyrrolyl -OMe 29 ~M 30 vinyl vi~yloxy vinyl 3.4 ~M 250 H BzO- H 16 ~M 50 -OMe BzO- BzO- 8.3 ~M 100 20-OMe -C(Me)=CH2 -OMe 24 ~M 160 -OEt -C(Me)=CH2 -OEt 2.5 ~M 160 -OMe -SMe -OMe 19 ~M 145 -SMe -ONe -S(O)Me 18 ~M 490 -OMeBr -O(CH2)2C~(CH2~2COOHO.04 ~M 525 COOH
-OMeBr -O(CH2)2CO(N-Asp) 2 ~M 4300 H-OMe -O(CH2)2CO(N-Gly) 9.3 ~M 80 -OMeBr -O(CH2)2CO(N-Gly) 10 ~M 8 Me -CH3, E~ - -CH2CH3, Bz - -CH2C6k6, N-~ly - -NHCH2COOH, N-Asp --NHCH(COOH)CH2COOH, ~- phenyl, Ac - -COCH3 Example 2 Proceeding as described in E~ample l above, the compounds listed below were assayed and found to exhibit high activity and selectivity, as set for~ in Table II:
A) 2,~diamino-5-(~benzylo~cybenzyl)pyTimidine;
Wo 92/084~l PCr/US91/08515 " 209~.jl8 B) 2,~dia~r.uno-5-(3,~dimethoxy-5-benzyloxybenzyl)pyrimidine:
C) 2,4 diarr.uno-5-(3,4 dibenzyloxy-5-methoxybenzyl)pyrirr.udine;
D) 2,~diamino-5-[3,5-dimethoxy-~(2-hydroxyprop-2-yl)benzyl~pyrilI~dine;
E) 2,~diamino-5-(3,5-dimethoxy- 1 N-pyrrolylbenzyl)pynmidine:
F) 2,1 diammo-5-(3,5-diethoxy-4-N-pyrrolylbenzyl)pyIimidine;
G) 2,1 diamino-5-(3~5-divinyl-4 vinyloxybenzyl)pyrimidine;
H) 2,4-diamino-5-[3-(l N-acetaminophenyl)sulfonaminoethoxy-4,5 dimeth-oxybenzyl]pyrim~dine;
I) 2,~dia~runo-5-[3-(4 aminophenyl)sulfonatninoethoxy-~methoxybenzyl]-pyri~udine;
J) 2,~diamino-5-[3-(4N-acetaminophenyl)sulfona n~noethoxy-~l bromo-5-methoxybenzyl]pyrimidine;
K) 2,1 diarnino-5-[3-(~aminophenyl~sulfonaminoethoxy-~bromo-5-methoxy-benzyl]pyrimidine;
L) 2,~diamino-5-[3,5-diethoxy-1 (propen-2-yl)benzyl]pyr~idine;
M) 2,~1 diamino-5-[3,5-dimethoxy-1 (propen-2-yl)benzyl]pyrimidine;
N~ 2,1 diarnino~5-(3,5-dimethoxy-~methylthiobenzyl)pyrimidine;
O) 2,~diamino-5-(3-methylsulfinyl-~methoxy-5-methylthiobenzyl)-pyrirnidine;
P) 2,4-diamino-5-[3-(4,6-dicarboxyhexyloxy)-4-bromo-5-methoxybenzyl]-pyrimidine;
Q) 2,4-diamino-5-[3-(3-carboxymethylamino-3-oxopropoxy)-l methoxy-benzyl]pynmidine;
R) 2,~diamino-5-(3-[3-(1,3-dicarboxypropyl)amino-3-oxopropoxy]-1 bromo-5-methoxybenzyl)pylimidi~e;
S) 2,~dian~no-5-[3,5-dimetho~cy-~((2-phenylsulfonyl)acetyl)benzyl]-pyrimidine;
T) 2,~diam~no-5-[3-am~no-4 methyl-5-(N-pyIrolyl)benzyl]pynmidine;
U) 2,4 diamino-5-(3,5-di-N-pyrrQlyl-4 methoxybenzyl)pyrimidine;
Wo 92/08461 PCr/US91/08~.15 2~9~18 V) 2,4-diamino-5-[3,5-di-methoxy-4 (3-hydrocarboxy-1-oxoprowlamino)-benzyl]pyri~udine;
W) 2,4-dian~ino-5-[3,5-dimethoxy-(~acetaminophenylsulfonarnino)benzyll-pyr~midine;
5 X) 2,~diamino-5-(3,5-dimethoxy-~propylbenzyl)pyrimidine;
Y) 2,4-diarnino-5-(3,5-dichloro-4-N-pyrrolylbenzyl)pyrimidine;
Z) 2,~diamino-5-[3,5-dimethoxy-4 (2-(2-(2-methoxy)ethoxy)ethoxy)ethoxy-benzyl]pyrimidine:
AA) 2,4-diarnino-5-[3-(3-benzyloxycarbonylmethylamino-3-oxopropoxy)-~
bromo-S-methoxybenzyl]pyrimidine;
BB) 2,4-diamino-5-[3-(3-carboxymethylamino-3-oxopropoxy)-4-bromo-5-methoxybenzyl]pyri~udine;
CC) 2,4-diamino-5-[3-methoxy-4-bromo-5-(4-methylaminobenzamidoethoxy)-benzyl]pyrimidine;
DD) 3-(2,1 diaminopyrimidin-5-ylmethyl)-8-(3,5-dimethoxyphenyl)-8-aza-bicyclo[3 .2.1 ]octane;
EE~ 2~I,3H-dihydro-5-(2,~diaminopy~imidin-5-ylmethyl)-6,7-dimethoxybenzo-furan;
FF) 5-(2,4-diaminopyrimidin-5-ylmethyl)-7-methoxy-8-bromo-1.2-benzopyran;
GG) 5-(2,~diaminopyrimidin-5-ylmethyl3-7,8-dimethoxy- 1 ,2-benzopyran;
HH) 2,~diamino-5-[3-phenyl-5-(3-methoxypropoxy)benzyl]pyr~Irudine:
II) 2,'1 diasnino-7-(3,5-dirnethoxybenzyl)pyrrolo[2,3-f~quinazoline;
JJ) 2,1 diamino-5-[~(4 methoxybutoxy)naphth-1-yl]pyrimidine;
KK) 2,4 diamin~5-(4,5,~tnrnethoxy-2,3 dihydroinden-1-yl)pynmidine;
LL) 2,2-dimethyl-5-(2,~diaminopyriIrLidin-S-ylmethyl)-7-methoxybenz[b]di-oxolane;
MM) 2,~diamino-5-(3,5-diethoxy-1 carboethoxybenzyl)pyrimidine; and NN) 2,1 diamino-5-(2,7-dimethylbenzpyrazol-5-yLmethyl)pyrirrudine.
WO 92/0&~1 PCT/US91/08515 2~3;~ 1;8 TABLE ~:
¦ Compound IC50 Selectivity ¦
_ _, A) 16.0 53.2 B) 13.9 3.3 C) 8.3 90.3 ¦ D) 24.8 343.5 E) 2.9 29.4 F) 2.4 312.4 G) 3.4 250.5 H) 2.4 248.5 I) 0.57 7.8 J) 3.7 29.5 ~) 0.97 12.1 L) 2.5 190.9 M) 24.0 159.8 N) lg.o 143.5 O) 17.5 486.8 P) 0.043 528.0 Q) 9.3 77.7 R) 2.0 4255.0 S) 14.8 36.8 T) 33.5 42.5 U) 4.5 47.7 V) 100.0 85.2 W) 31.0 14.3 X) 20.0 42.6 Y) 10.9 53.2 Z) 36.0 33.1 WO92~08461 PCT/US91/08,C.15 ~9 -~8 TABLE II (continued):
Compound IC50Selectivity AA) 7.9 14.4 BB) 10.0 13.1 CC) 7.8 14.6 DD) 0.016 31.0 EE) 12.9 26.4 FF) 5.9 6.9 GG) 11.5 266.7 HH) 12.2 11.2 II) 0.039 1.2 JJ) 80.0 106.5 KK) 87.0 97.9 I LL) 13.9 15.0 NM) 8.3 63.6 I NN) 18.0 10.7
Claims (35)
1. A method for treating fungal infection in a mammal which method comprises:
administering to a mammal infected with a fungal pathogen an effective amount of a compound of formula I:
wherein R1 is 3-R3-4-R4-5-R5-benzyl or (N-R6)-8-azabicyclo[3.2.1]oct-3-yl;
and R2 is H, or R1 and R2 together form ;
where R3 and R5 are independently selected from the group consist-ing of H. lower alkoxy, lower alkylthio, lower alkylsulfinyl, vinyl, carboxy-lower alkyl, carboxy-lower alkoxy, dicarboxy-lower alkyl, dicarboxy-lower alkoxy, aryl-lower alkoxy, arylsulfonyl-lower alkoxy, arylsulfamido-lower alkoxy, and radicals of formula -O(CH2)n-COR8, where n is an integer from 0 to 6 and R8 is an amino acid;
R4 is selected from the group consisting of lower alkoxy, aryl-lower alkoxy, lower alkylthio, halo, lower alkenyl, lower alkenyloxy, and pyrrolyl;
with the proviso that R3, R4, and R5 are not simultaneously methoxy;
R6 is selected from the group consisting of unsubstituted aryl and aryl substituted with one to three radicals selected from the group consisting of halo, lower alkyl, lower alkoxy, lower alkylthio, carboxy, carbamido, carboxy-lower alkyl, and carbamido-lower alkyl; and R7 is selected from the group consisting of aryl and aryl-lower alkyl, where aryl may be substituted with one to three radicals selected from the groupconsisting of halo, lower alkyl, lower alkoxy, lower alkylthio, carboxy, carbamido, carboxy-lower alkyl, and carbamido-lower alkyl;
and lower alkyl esters, amides thereof, and pharmaceutically acceptable addition salts.
administering to a mammal infected with a fungal pathogen an effective amount of a compound of formula I:
wherein R1 is 3-R3-4-R4-5-R5-benzyl or (N-R6)-8-azabicyclo[3.2.1]oct-3-yl;
and R2 is H, or R1 and R2 together form ;
where R3 and R5 are independently selected from the group consist-ing of H. lower alkoxy, lower alkylthio, lower alkylsulfinyl, vinyl, carboxy-lower alkyl, carboxy-lower alkoxy, dicarboxy-lower alkyl, dicarboxy-lower alkoxy, aryl-lower alkoxy, arylsulfonyl-lower alkoxy, arylsulfamido-lower alkoxy, and radicals of formula -O(CH2)n-COR8, where n is an integer from 0 to 6 and R8 is an amino acid;
R4 is selected from the group consisting of lower alkoxy, aryl-lower alkoxy, lower alkylthio, halo, lower alkenyl, lower alkenyloxy, and pyrrolyl;
with the proviso that R3, R4, and R5 are not simultaneously methoxy;
R6 is selected from the group consisting of unsubstituted aryl and aryl substituted with one to three radicals selected from the group consisting of halo, lower alkyl, lower alkoxy, lower alkylthio, carboxy, carbamido, carboxy-lower alkyl, and carbamido-lower alkyl; and R7 is selected from the group consisting of aryl and aryl-lower alkyl, where aryl may be substituted with one to three radicals selected from the groupconsisting of halo, lower alkyl, lower alkoxy, lower alkylthio, carboxy, carbamido, carboxy-lower alkyl, and carbamido-lower alkyl;
and lower alkyl esters, amides thereof, and pharmaceutically acceptable addition salts.
2. The method of claim 1, wherein R2 is H and R1 is 3-R3-4-R4-5-R5-benzyl.
3. The method of claim 2, wherein R3 is methoxy.
4. The method of claim 3, wherein R4 is bromo.
5. The method of claim 4, wherein R5 is -O(CH2)2CH(COOH)(CH2)2COOH, or a pharmaceutically acceptable mono- or diester thereof.
6. The method of claim 4, wherein R5 is -O(CH2)n-COR5.
7. The method of claim 6, wherein n is 2 and R8 is -NHCH(COOH)CH2CH2COOH or a mono- or diester thereof.
8. The method of claim 6, wherein n is 2 and R8 is -NHCH2COOH or an ester thereof.
9. The method of claim 3 wherein R5 is methoxy.
10. The method of claim 9, wherein R4 is 2-hydroxyprop-2-yl.
11. The method of claim 9, wherein R4 is propen-2-yl.
12. The method of claim 9, wherein R4 is N-pyrrolyl.
13. The method of claim 9, wherein R4 is methylthio.
14. The method of claim 3, wherein R4 is benzyloxy.
15. The method of claim 14, wherein R5 is benzyloxy.
16. The method of claim 3, wherein R4 is methoxy, and R5 is -OCH2CH2NHSO2(C6H6)NHCOCH3.
17. The method of claim 2 wherein R3 is H.
18. The method of claim 9 wherein R4 is methoxy.
19. The method of claim 10 wherein R5 is -O(CH2)n-COR8.
where n is 2 and R8 is -NHCH2COOH or an ester thereof.
where n is 2 and R8 is -NHCH2COOH or an ester thereof.
20. The method of claim 9, wherein R4 is benzyloxy and R5 is H.
21. The method of claim 2, where R3 and R5 are each ethoxy.
22. The method of claim 21, wherein R4 is pyrrolyl.
23. The method of claim 21, wherein R4 is propen-2-yl.
24. The method of claim 2, where R3 and R5 are each vinyl.
25. The method of claim 2, where R3 is methylthio, R4 is methoxy, and R5 is methylsulfinyl.
26. The method of claim 1, wherein R2 is H and R1 is (N-R6)-8-azabicyclo[3.2.1]oct-3-yl.
27. The method of claim 26, wherein R6 is selected from the group consisting of 2-naphthyl, 3,5-dimethoxyphenyl, and 4-carboethoxyphenyl.
28. The method of claim 1, wherein R1 and R2 together form
29. The method of claim 28, wherein R7 is benzyl.
30. The method of claim 28 where R7 is 3,5-dimethoxyphenyl.
31. The method of claim 2 where R3 and R5 are propoxy and R4 is N-pyrrolyl.
32. The method of claim 1 wherein said fungal pathogen is Pneumocystis carinii.
33. The method of claim 1, further comprising coadminstering a dihydropteroate synthase inhibitor.
34. The method of claim 33, wherein said dihydropteroate synthase inhibitor is selected from the group consisting of dapsone and sulfa drugs.
35. The method of claim 1, wherein said compound of formula I is selected from the group consisting of 2,4-diamino-5-(4-benzyloxy-benzyl)pyrimidine; 2,4-diamino-5-(3,4-dimethoxy-5-benzyloxybenzyl)pyrimidine;
2,4-diamino-5-(3,4-dibenzyloxy-5-methoxybenzyl)pyrimidine; 2,4-diamino-5-[3,5-dimethoxy-4-(2-hydroxyprop-2-yl)benzyl]pyrimidine; 2,4-diamino-5-(3,5-dimethoxy-4-N-pyrrolylbenzyl)pyrimidine; 2,4-diamino-5-(3,5-diethoxy-4-N-pyrrolylbenzyl)pyrimidine; 2,4-diamino-5-(3,5-divinyl-4-vinyloxybenzyl)-pyrimidine; 2,4-diamino-5-[3-(4-N-acetaminophenyl)sulfonaminoeethoxy-4,5-di-methoxybenzyl]pyrimidine; 2,4-diamino-5-[3-(4-aminophenyl)sulfonaminoethoxy-4-methoxybenzyl]pyrimidine; 2,4 diamino-5-[3-(4-N-acetaminophenyl)sulfonaminoethoxy-4-bromo-5-methoxybenzyl]pyrimidine; 2,4-di-amino-5-[3-(4-aminophenyl)sulfonaminoethoxy-4-bromo-5-methoxybenzyl]-pyrimidine; 2,4-diamino-5-[3,5-diethoxy-4-(propen-2-yl)benzyl]pyrimidine; 2,4-diamino-5-[3,5-dimethoxy-4-(propen-2-yl)benzyl]pyrimidine; 2,4-diamino-5-(3,5-dimethoxy-4-methylthiobenzyl)pyrimidine; 2,4-diamino-5-(3-methylsulfinyl-4-methoxy-5-methylthiobenzyl)pyrimidine; 2,4-diamino-5-[3-(4,6-dicarboxyhexyloxy)-4-bromo-5-methoxybenzyl]pyrimidine; 2,4-diamino-5-[3-(3-carboxymethylamino-3-oxopropoxy)-4-methoxybenzyl]pyrimidine; 2,4-diamino-5-(3-[3-(1,3-dicarboxypropyl)amino-3-oxopropoxy]-4-bromo-5-methoxybenzyl)-pyrimidine; 2,4-diamino-5-[3,5-dimethoxy-4-((2-phenylsulfonyl)acetyl)benzyl]-pyrimidine; 2,4-diamino-5-[3-amino-4-methyl-5-(N-pyrrolyl)benzyl]pyrimidine;
2,4-diamino-5-(3,5-di-N-pyrrolyl-4-methoxybenzyl)pyrimidine; 2,4-diamino-5-[3,5-di-methoxy-4-(3-hydrocarboxy-1-oxopropylamino)benzyl]pyrimidine; 2,4-diamino-5-[3,5-dimethoxy-(4-acetaminophenylsulfonamino)benzyl]pyrimidine; 2,4-diamino-5-(3,5-dimethoxy-4-propylbenzyl)pyrimidine; 2,4-diamino-5-(3,5-dichloro-4-N-pyrrolylbenzyl)pyrimidine; 2,4-diamino-5-[3,5-dimethoxy-4-(2-(2-(2-methoxy)-ethoxy)ethoxy)ethoxybenzyl]pyrimidine; 2,4-diamino-5-[3-(3-benzyloxycarbonyl-methylamino-3-oxopropoxy)-4-bromo-5-methoxybenzyl]pyrimidine; 2,4-diamino-5-[3-(3-carboxymethylamino-3-oxopropoxy)-4-bromo-5-methoxybenzyl]pyrimidine;
2,4-diamino-5-[3-methoxy-4-bromo-5-(4-methylaminobenzamidoethoxy)benzyl]pyr-imidine; 3-(2,4-diaminopyrimidin-5-ylmethyl)-8-(3,5-dimethoxyphenyl)-8-aza-bicyclo[3.2.1]octane; 2H,3H-dihydro-5-(2,4-diaminopyrimidin-5-ylmethyl)-6,7-di-methoxybenzofuran; 5-(2,4-diaminopyrimidin-5-ylmethyl)-7-methoxy-8-bromo-1,2-benzopyran; 5-(2,4-diaminopyrimidin-5-ylmethyl)-7,8-dimethoxy-1,2-benzopyran;
2,4-diamino-5-[3-phenyl-5-(3-methoxypropoxy)benzyl]pyrimidine; 2,4-diamino-7-(3,5-dimethoxybenzyl)pyrrolo[2,3-f]quinazoline; 2,4-diamino-5-[6-(4-methoxy-butoxy)naphth-1-yl]pyrimidine; 2,4 diamino-5-(4,5,6-trimethoxy-2,3-dihydroinden-1-yl)pyrimidine; 2,2-dimethyl-5-(2,4-diaminopyrimidin-5-ylmethyl)-7-methoxy-benz[b]dioxolane; 2,4-diamino-5-(3,5-diethoxy-4-carboethoxybenzyl)pyrimidine;
and 2,4 diamino-5-(2,7-dimethylbenzpyrazol-5-ylmethyl)pyrimidine.
2,4-diamino-5-(3,4-dibenzyloxy-5-methoxybenzyl)pyrimidine; 2,4-diamino-5-[3,5-dimethoxy-4-(2-hydroxyprop-2-yl)benzyl]pyrimidine; 2,4-diamino-5-(3,5-dimethoxy-4-N-pyrrolylbenzyl)pyrimidine; 2,4-diamino-5-(3,5-diethoxy-4-N-pyrrolylbenzyl)pyrimidine; 2,4-diamino-5-(3,5-divinyl-4-vinyloxybenzyl)-pyrimidine; 2,4-diamino-5-[3-(4-N-acetaminophenyl)sulfonaminoeethoxy-4,5-di-methoxybenzyl]pyrimidine; 2,4-diamino-5-[3-(4-aminophenyl)sulfonaminoethoxy-4-methoxybenzyl]pyrimidine; 2,4 diamino-5-[3-(4-N-acetaminophenyl)sulfonaminoethoxy-4-bromo-5-methoxybenzyl]pyrimidine; 2,4-di-amino-5-[3-(4-aminophenyl)sulfonaminoethoxy-4-bromo-5-methoxybenzyl]-pyrimidine; 2,4-diamino-5-[3,5-diethoxy-4-(propen-2-yl)benzyl]pyrimidine; 2,4-diamino-5-[3,5-dimethoxy-4-(propen-2-yl)benzyl]pyrimidine; 2,4-diamino-5-(3,5-dimethoxy-4-methylthiobenzyl)pyrimidine; 2,4-diamino-5-(3-methylsulfinyl-4-methoxy-5-methylthiobenzyl)pyrimidine; 2,4-diamino-5-[3-(4,6-dicarboxyhexyloxy)-4-bromo-5-methoxybenzyl]pyrimidine; 2,4-diamino-5-[3-(3-carboxymethylamino-3-oxopropoxy)-4-methoxybenzyl]pyrimidine; 2,4-diamino-5-(3-[3-(1,3-dicarboxypropyl)amino-3-oxopropoxy]-4-bromo-5-methoxybenzyl)-pyrimidine; 2,4-diamino-5-[3,5-dimethoxy-4-((2-phenylsulfonyl)acetyl)benzyl]-pyrimidine; 2,4-diamino-5-[3-amino-4-methyl-5-(N-pyrrolyl)benzyl]pyrimidine;
2,4-diamino-5-(3,5-di-N-pyrrolyl-4-methoxybenzyl)pyrimidine; 2,4-diamino-5-[3,5-di-methoxy-4-(3-hydrocarboxy-1-oxopropylamino)benzyl]pyrimidine; 2,4-diamino-5-[3,5-dimethoxy-(4-acetaminophenylsulfonamino)benzyl]pyrimidine; 2,4-diamino-5-(3,5-dimethoxy-4-propylbenzyl)pyrimidine; 2,4-diamino-5-(3,5-dichloro-4-N-pyrrolylbenzyl)pyrimidine; 2,4-diamino-5-[3,5-dimethoxy-4-(2-(2-(2-methoxy)-ethoxy)ethoxy)ethoxybenzyl]pyrimidine; 2,4-diamino-5-[3-(3-benzyloxycarbonyl-methylamino-3-oxopropoxy)-4-bromo-5-methoxybenzyl]pyrimidine; 2,4-diamino-5-[3-(3-carboxymethylamino-3-oxopropoxy)-4-bromo-5-methoxybenzyl]pyrimidine;
2,4-diamino-5-[3-methoxy-4-bromo-5-(4-methylaminobenzamidoethoxy)benzyl]pyr-imidine; 3-(2,4-diaminopyrimidin-5-ylmethyl)-8-(3,5-dimethoxyphenyl)-8-aza-bicyclo[3.2.1]octane; 2H,3H-dihydro-5-(2,4-diaminopyrimidin-5-ylmethyl)-6,7-di-methoxybenzofuran; 5-(2,4-diaminopyrimidin-5-ylmethyl)-7-methoxy-8-bromo-1,2-benzopyran; 5-(2,4-diaminopyrimidin-5-ylmethyl)-7,8-dimethoxy-1,2-benzopyran;
2,4-diamino-5-[3-phenyl-5-(3-methoxypropoxy)benzyl]pyrimidine; 2,4-diamino-7-(3,5-dimethoxybenzyl)pyrrolo[2,3-f]quinazoline; 2,4-diamino-5-[6-(4-methoxy-butoxy)naphth-1-yl]pyrimidine; 2,4 diamino-5-(4,5,6-trimethoxy-2,3-dihydroinden-1-yl)pyrimidine; 2,2-dimethyl-5-(2,4-diaminopyrimidin-5-ylmethyl)-7-methoxy-benz[b]dioxolane; 2,4-diamino-5-(3,5-diethoxy-4-carboethoxybenzyl)pyrimidine;
and 2,4 diamino-5-(2,7-dimethylbenzpyrazol-5-ylmethyl)pyrimidine.
Applications Claiming Priority (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US61361990A | 1990-11-14 | 1990-11-14 | |
| US613,619 | 1990-11-14 | ||
| US73069191A | 1991-07-16 | 1991-07-16 | |
| US730,691 | 1991-07-16 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2095518A1 true CA2095518A1 (en) | 1992-05-15 |
Family
ID=27087063
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002095518A Abandoned CA2095518A1 (en) | 1990-11-14 | 1991-11-14 | Specific inhibition of dihydrofolate reductase and compounds therefor |
Country Status (7)
| Country | Link |
|---|---|
| EP (1) | EP0639075A1 (en) |
| JP (1) | JPH07509215A (en) |
| AU (1) | AU657348B2 (en) |
| CA (1) | CA2095518A1 (en) |
| IE (1) | IE913974A1 (en) |
| PT (1) | PT99514B (en) |
| WO (1) | WO1992008461A1 (en) |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5721242A (en) * | 1993-06-17 | 1998-02-24 | Hoffmann-La Roche Inc. | Antibiotic combination |
| KR100417508B1 (en) * | 1994-11-24 | 2004-03-18 | 바실리어 파마슈티카 아게 | Novel benzyl pyrimidines |
| TW438796B (en) * | 1996-05-15 | 2001-06-07 | Hoffmann La Roche | 2,4-diaminopyrimidine derivatives, the manufacture process thereof, and the antibiotically-active pharmaceutical composition containing the same |
| AU6203098A (en) * | 1997-02-24 | 1998-09-18 | Barry J. Barclay | Synergistically acting anti-fungal agents that are inhibitors of folate metabolism, especially methotrexate and sulfamethoxazole |
| HUP0201333A3 (en) | 1999-05-24 | 2004-11-29 | Sankyo Company Ltd Chuo Ku | Medicinal compositions containing anti-fas antibody |
| US8853228B2 (en) * | 2007-06-04 | 2014-10-07 | University Of Connecticut | Heterocyclic analogs of propargyl-linked inhibitors of dihydrofolate reductase |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US2658897A (en) * | 1951-06-27 | 1953-11-10 | Burroughs Wellcome Co | 2, 4-diamino-5-benzyl pyrimidines |
| FR1090394A (en) * | 1953-03-11 | 1955-03-30 | Wellcome Found | Pyrimidine derivatives and their manufacture |
| US2852450A (en) * | 1954-06-10 | 1958-09-16 | Donnelley & Sons Co | Method of copper plating |
| FR1292920A (en) * | 1959-04-18 | 1962-05-11 | Wellcome Found | Pyrimidine derivatives and their preparation |
| GB1142654A (en) * | 1965-10-28 | 1969-02-12 | Wellcome Found | Cyano-acetals and their use in benzylpyrimidine synthesis |
| FR94837E (en) * | 1966-02-19 | 1969-11-28 | Wellcome Found | Process for the preparation of pyrimidine derivatives. |
| GB1181657A (en) * | 1966-03-31 | 1970-02-18 | Ici Ltd | Pyrimidine Derivatives and Compositions Containing them |
| BE793253A (en) * | 1971-12-23 | 1973-04-16 | Lepetit Spa | NEW BACTERICIDE COMPOSITIONS AND THEIR USE |
| CH591457A5 (en) * | 1973-11-08 | 1977-09-15 | Hoffmann La Roche | |
| DE2617967C3 (en) * | 1976-04-24 | 1978-11-02 | Nordmark-Werke Gmbh, 2000 Hamburg | Process for the preparation of 2,4-diamino-5-benzylpyrimidines |
| IE45427B1 (en) * | 1976-07-09 | 1982-08-25 | American Home Prod | Pyrrold 3,2if quinazoline-1,3-diamine and related compounds |
| LU77268A1 (en) * | 1977-05-05 | 1979-01-18 | ||
| LU77269A1 (en) * | 1977-05-05 | 1979-01-18 | ||
| GR71725B (en) * | 1977-11-10 | 1983-06-22 | Hoffmann La Roche | |
| US4233445A (en) * | 1979-07-05 | 1980-11-11 | American Home Products Corporation | 7-(Substituted)-7H-pyrrolo[3.2-F]-quinazoline-1,3-diamines |
| US4258045A (en) * | 1979-11-30 | 1981-03-24 | Merck & Co., Inc. | Inhibitor of dihydrofolate reductase |
| OA07362A (en) * | 1980-10-27 | 1984-06-30 | May & Baker Ltd | Pyrimidine derivatives, process for their preparation and compositions containing them. |
| EP0054756B1 (en) * | 1980-11-27 | 1986-07-23 | The Wellcome Foundation Limited | Antibacterial benzylpyrimidines |
| US4451466A (en) * | 1982-07-02 | 1984-05-29 | Shell Oil Company | Use of pyrroloquinazolinediamines as pesticides |
| CA1244028A (en) * | 1983-04-14 | 1988-11-01 | Hans Maag | Pyrimidine derivatives |
| EP0139613A1 (en) * | 1983-08-29 | 1985-05-02 | Ciba-Geigy Ag | N-(2-nitrophenyl)-4-aminopyrimidine derivatives, their preparation and use |
| US4501890A (en) * | 1983-09-26 | 1985-02-26 | Eli Lilly And Company | Trans-(±)-2,4,6-substituted-5,5a,6,7,8,9,9a,10-octahydro-pyrimido[4,5-g]quinolines |
| DE3417264A1 (en) * | 1984-05-10 | 1985-11-14 | Bayer Ag, 5090 Leverkusen | NEW 2,4-DIAMINO-6-HALOGEN-5-ALKYLTHIO-PYRIMIDINE |
| DE3701277A1 (en) * | 1987-01-17 | 1988-07-28 | Boehringer Mannheim Gmbh | NEW TRICYCLIC BENZIMIDAZOLES, METHOD FOR THEIR PRODUCTION AND USE AS MEDICINAL PRODUCTS |
| US4996198A (en) * | 1988-07-11 | 1991-02-26 | Hoffmann-La Roche Inc. | Anticoccidial composition |
-
1991
- 1991-11-14 PT PT99514A patent/PT99514B/en not_active IP Right Cessation
- 1991-11-14 CA CA002095518A patent/CA2095518A1/en not_active Abandoned
- 1991-11-14 WO PCT/US1991/008515 patent/WO1992008461A1/en not_active Application Discontinuation
- 1991-11-14 EP EP92900837A patent/EP0639075A1/en not_active Ceased
- 1991-11-14 IE IE397491A patent/IE913974A1/en not_active Application Discontinuation
- 1991-11-14 JP JP4502293A patent/JPH07509215A/en active Pending
-
1992
- 1992-08-31 AU AU22027/92A patent/AU657348B2/en not_active Ceased
Also Published As
| Publication number | Publication date |
|---|---|
| WO1992008461A1 (en) | 1992-05-29 |
| PT99514A (en) | 1992-10-30 |
| JPH07509215A (en) | 1995-10-12 |
| AU2202792A (en) | 1994-04-28 |
| AU657348B2 (en) | 1995-03-09 |
| IE913974A1 (en) | 1992-05-20 |
| EP0639075A1 (en) | 1995-02-22 |
| PT99514B (en) | 1999-04-30 |
| EP0639075A4 (en) | 1993-09-29 |
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| Date | Code | Title | Description |
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| FZDE | Dead |