[go: up one dir, main page]

WO1996029329A1 - Urees cycliques inhibitrices de la protease de vih - Google Patents

Urees cycliques inhibitrices de la protease de vih Download PDF

Info

Publication number
WO1996029329A1
WO1996029329A1 PCT/US1996/003426 US9603426W WO9629329A1 WO 1996029329 A1 WO1996029329 A1 WO 1996029329A1 US 9603426 W US9603426 W US 9603426W WO 9629329 A1 WO9629329 A1 WO 9629329A1
Authority
WO
WIPO (PCT)
Prior art keywords
substituted
benzyl
compound
indazolylmethyl
aminobenzyl
Prior art date
Application number
PCT/US1996/003426
Other languages
English (en)
Inventor
Prabhakar Kondaji Jadhav
Original Assignee
The Du Pont Merck Pharmaceutical Company
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from US08/613,554 external-priority patent/US5683999A/en
Application filed by The Du Pont Merck Pharmaceutical Company filed Critical The Du Pont Merck Pharmaceutical Company
Priority to AU53100/96A priority Critical patent/AU5310096A/en
Priority to EP96909680A priority patent/EP0815108A1/fr
Publication of WO1996029329A1 publication Critical patent/WO1996029329A1/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D243/00—Heterocyclic compounds containing seven-membered rings having two nitrogen atoms as the only ring hetero atoms
    • C07D243/04—Heterocyclic compounds containing seven-membered rings having two nitrogen atoms as the only ring hetero atoms having the nitrogen atoms in positions 1 and 3
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
    • C07D405/12—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/14—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
    • C07D409/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
    • C07D413/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
    • C07D417/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing three or more hetero rings

Definitions

  • This invention relates to substituted cyclic ureas and derivatives thereof, including compounds of
  • the present invention also relates to pharmaceutical compositions comprising such compounds and to methods of using these compounds for the
  • the present invention also relates to the use of such compounds in processes for the identification of HIV protease inhibitors and for the inhibition or detection of HIV in a bodily fluid sample.
  • Retroviral proteases encode a protease that is responsible for the proteolytic processing of one or more polyprotein precursors such as the pol and gag gene products. Retroviral proteases most commonly process the gag precursor into the core proteins, and also process the pol precursor into reverse transcriptase (RT) and retroviral protease.
  • RT reverse transcriptase
  • polyproteins by the retroviral protease is necessary for the assembly of the infectious virions.
  • Protease-defective virus leads to the production of immature core forms which lack infectivity. Therefore, retroviral protease inhibition provides an attractive target for antiviral therapy.
  • HIV Human immunodeficiency virus
  • HlV-1 HlV-1
  • type-2 type-2
  • HIV seropositive individuals are initially asymptomatic but typically develop AIDS related complex (ARC) followed by AIDS.
  • ARC AIDS related complex
  • Affected individuals exhibit severe immunosuppression which predisposes them to debilitating and ultimately fatal opportunistic infections.
  • AIDS is characaterized by a progressive depletion of T lymphocytes, particularly the helper-inducer subset having a CD4 surface antigen (CD4 + ).
  • CD4 + lymphocytes are responsible for the induction of numerous functions of the human immune system.
  • RT HIV reverse transcriptase
  • nucleoside analogs including 3 ' - azido- 3 ' - deoxythymidine (AZT, zidovidine) and 2',3'-dideoxy nucleosides such as 2',3'-dideoxyinosine (DDI, ddI) and
  • DDC 2',3'-dideoxycytidine
  • AZT is known to frequently cause adverse side effects, including causing bone marrow suppression resulting in anemia, leukopenia and thrombocytopenia. Also, AZT- resistant HIV strains have been observed in patients following AZT treatment.
  • HIV protease encoded by HIV is critical for replication of the virus. HIV protease is responsible for specific cleavages of the viral gag/pol gene products, which are precursors of essential viral structural proteins and essential enzymes including RT, integrase, and the protease itself (Ratner et al.,
  • PCT Patent Applications WO 93/07128 published April 15, 1993 and WO 94/19329 published September 1, 1994 disclose cyclic ureas as inhibitors of HIV protease.
  • the compounds of the invention are of low molecular weight and may, therefore, provide good oral absorption properties in mammals.
  • the compounds of the present invention provide improved antiviral potency, including increased potency against various HIV mutants which are known to be significantly less susceptible to other known inhibitors of HIV protease.
  • This invention provides novel substituted cyclic urea compounds and derivatives thereof, of formula (I) (described below) which are useful as inhibitors of human immunodeficiency virus (HIV).
  • the compounds of the present invention inhibit HIV protease and thereby inhibit HIV replication.
  • the present invention also includes pharmaceutical compositions comprising such compounds of formula (I) and methods of using such compounds for the treatment of HIV infection in a patient.
  • kits comprising one or more containers containing pharmaceutical dosage units comprising a compound of formula (I), for the treatment of HIV infection.
  • the present invention also includes methods of inhibiting HIV or treating HIV infection by
  • a compound of formula (I) in combination with one or more second therapeutic agents selected from other inhibitors of HIV and/or therapeutic agents for the treatment of HIV-mediated disease conditions.
  • the present invention includes methods of using such compounds for the detection and inhibition of HIV in a sample, including a bodily fluid sample, which containing HIV.
  • This invention provides novel substituted cyclic urea compounds and derivatives thereof, of formula (I) (described below) which are useful as inhibitors of human immunodeficiency virus (HIV).
  • the compounds of the present invention inhibit HIV protease and thereby inhibit HIV replication.
  • the present invention also includes pharmaceutical compositions comprising such compounds of formula (I) and methods of using such compounds for the treatment of HIV infection in a patient.
  • kits comprising one or more containers containing pharmaceutical dosage units comprising a compound of formula (I), for the treatment of HIV infection.
  • the present invention also includes methods of inhibiting HIV or treating HIV infection by
  • a compound of formula (I) in combination with one or more second therapeutic agents selected from other inhibitors of HIV and/or therapeutic agents for the treatment of HIV-mediated disease conditions.
  • the present invention includes methods of using such compounds for the detection and inhibition of HIV in a sample, including a bodily fluid sample, which containing HIV.
  • R 1 is -CH 2 -X-Y-Z
  • X is selected from:
  • heterocycle selected from the group consisting of pyridine, thiophene, furan, thiazole, indazole, thieno[2,3-c]pyrazole and thieno[3,2-c]pyrazole, said heterocycle being substituted with 0-2 R 7 ;
  • Y is selected from:
  • Z is selected from:
  • 2-pyrimidinyl substituted with 0-1 R 5 and 0-1 R 6 ; 2-pyrazinyl, substituted with 0-1 R 5 and 0-1 R 6 ; 4-pyrimidon-2-yl or 2-pyrimidon-4-yl, substituted with 0-1 R 5 and 0-1 R 6 ;
  • Q is O, S or NH
  • Q 1 is CR 5 or N
  • R 2 is selected from:
  • 5- or 6-indazolylmethyl 5-benzotriazolylmethyl, 5- benzoxazolonylmethyl, 6-benzoxazolonylmethyl, 5-benzimidazolylmethyl, 5-benzoxazolylmethyl, or 5-benzisoxazolylmethyl, said indazolyl, benzimidazolyl, benzoxazolyl and
  • Y 1 is selected from:
  • Z 1 is selected from:
  • 2-pyrimidinyl substituted with 0-1 R 5 and 0-1 R 6 ; 2-pyrazinyl, substituted with 0-1 R 5 and 0-1 R 6 ; 4-pyrimidon-2-yl or 2 -pyrimidon-4 -yl, substituted with 0-1 R 5 and 0-1 R 6 ;
  • R 3 and R 4 are independently selected from:
  • 3-methoxybenzyl 4-methoxybenzyl, ethyl, isobutyl, cyclohexylmethyl, n-hexyl, 4-nitrobenzyl, 4- aminobenzyl, 3 -hydroxybenzyl, 4-hydroxybenzyl, 4- benzyloxybenzyl, 4-thiomethylbenzyl, 2- thienylmethyl, 3-thienylmethyl, 2-thiazolylmethyl, 4-pyridylmethyl 3 , 4-methylenedioxybenzyl, 3,4- ethylenedioxybenzyl, 1,4-benzoxazin-6-yl-methyl, 4-N,N- dimethylaminobenzyl or 2-naphthylmethyl;
  • R 5 and R 6 are independently selected from:
  • alkylcarbonyl benzyloxy, C 3 -C 6 cycloalkoxy, or phenyl, said phenyl being optionally substituted with Cl, F, Br, -CN, -NO 2 , -NR 11 R 12 , -CF 3 , C ! -C 4 alkyl, C 1 -C 4 alkoxy or -OH, or
  • heterocyclic ring system said carbocyclic or heterocyclic ring system being optionally substituted with 1-3 groups independently selected from: Cl, F, Br, -CN, -NO 2 , -CF 3 , -CO 2 R 8 , -COR 8 , -OCOR 8 , -NR 11 R 12 , -CONHR 8 , C 1 -C 4 alkyl, C 1 -C 4 alkoxy or -OH;
  • R 7 is selected from: hydrogen, halogen, -CN, -NO 2 , -OR 8 , C 1 -C 4 haloalkyl, C 2 -C 4 haloalkoxy, -CO 2 R 8 , -COR 8 , -CONHR 8 , -NR 11 R 12 , -OCOR 8 , C 1 -C 6 alkyl, phenyl substituted with 0-2 R 10 , or
  • R 8 is hydrogen, C 1 -C 4 alkyl, phenyl substituted with 0-2 R 10 , or
  • R 9 is selected from: C 1 -C 3 alkyl, OR 8 , NR 11 R 12 ,
  • R 11 is selected from: hydrogen or C 1 -C 4 alkyl
  • R 12 is selected from: hydrogen, C 1 -C 4 alkyl, glycinyl, alanyl, alanyl-alanyl, phenylglycinyl, phenylalanyl or N-methylglycinyl, N,N-dimethylglycinyl;
  • R 13 is selected from: hydrogen, phenyl, benzyl, C 1 -C 6 alkyl or C 3 -C 6 alkoxyalkyl.
  • the present invention includes compounds of formula (I), of formula (II) :
  • R 1 is ;
  • Y is selected from:
  • Z is a heterocycle selected from:
  • heterocycle being substituted with 0-1 R 5 and 0-1 R 6; selected from:
  • R 1 -CH 2 -X-Y 1 -Z, cyclopropylmethyl, allyl, 3,3- dimethylallyl, 2-methylallyl, n-propyl, 3- methylbutyl, 2-methylpropyl, n-butyl, n- pentyl, iso-pentyl, 4,4,4-trifluorobutyl, 3- methoxypropyl, benzyl, 2 -naphthylmethyl, 3- methoxybenzyl, 4-methoxybenzyl,
  • 3-N,N-dipropylaminobenzyl 3-N-methylaminobenzyl, 4-N-methylaminobenzyl, 4-N-ethylaminobenzyl, 4-N,N-dimethylaminobenzyl,
  • Y 1 is selected from:
  • R 5 and R 6 are independently selected from:
  • R 7 is selected from: hydrogen, halogen, -CN, - ⁇ O 2 , -OH, CF 3 , OCH 3 , CO 2 H, CO 2 R 8 , COR 8 or C 1 -C 6 alkyl; R 8 is H or C 1 -C 4 alkyl.
  • Preferred compounds of the present invention include compounds of formula (II), or a pharmaceutically acceptable salt or prodrug form thereof, wherein: is ;
  • Y is -(CH 2 ) n O- or -(CH 2 ) n CONH-; n is 0; m is 1-2; Z is a heterocycle selected from:
  • R 2 is selected from:
  • R 1 -CH 2 -X-Y 1 -Z, cyclopropylmethyl, 3,3- dimethylallyl, 2-methylallyl, 3-methylbutyl, 2- methylpropyl, n-butyl, n-pentyl, iso-pentyl, 4,4,4- trifluorobutyl, 3-methoxypropyl, benzyl,
  • 3-N,N-dipropylaminobenzyl 3-N-methylaminobenzyl, 4-N-methylaminobenzyl, 4-N-ethylaminobenzyl, 4-N,N-dimethylaminobenzyl, 4-N-butylaminobenzyl, 3-(2-pyridylmethyl)aminobenzyl, 3- (carboethoxymethyl)aminobenzyl, 3- (ethoxycarbonyl)aminobenzyl, 3-amino-4-fluoro-benzyl, 4-fluorobenzyl, 4-chlorobenzyl, 3-azido-4-fluoro-benzyl, 3-trifluorobenzyl, 2,4-difluorobenzyl, 3-formaldoximebenzyl,
  • Y 1 is selected from:
  • R 5 and R 6 are independently selected from: halogen, NO 2 , CN, CF 3, C 1 -C 6 alkyl, or phenyl, said phenyl being optionally substituted with 1-3 groups
  • R 7 is selected from: hydrogen, halogen, -CN, -NO 2 , -OH, CF 3 , OCH 3 , or C 1 -C 6 alkyl.
  • Preferred compounds of the present invention include compounds of formula (II), or a pharmaceutically acceptable salt or prodrug form thereof, wherein: R 1 is ;
  • R 2 is selected from:
  • R 1 cyclopropylmethyl, 3,3-dimethylallyl, 2- methylallyl, 3-methylbutyl, 2-methylpropyl,
  • 3-N-ethylaminobenzyl 3-N,N-dimethylaminobenzyl, 3-N-methylaminobenzyl, 4-N-methylaminobenzyl, 3- hydroxymethylbenzyl, 4-hydroxymethylbenzyl, 2- naphthylmethyl, 3-(2-pyridylmethyl)aminobenzyl, 3- amino-4-fluoro-benzyl, 4-fluorobenzyl, 4- chlorobenzyl, 3-trifluorobenzyl,
  • Preferred compounds of the present invention include compounds of formula (I), or a pharmaceutically acceptable salt or prodrug form thereof, selected from:
  • Preferred compounds of the present invention include compounds, or a pharmaceutically acceptable salt form or prodrug form thereof, selected from:
  • the present invention also provides a
  • composition comprising a pharmaceutically acceptable carrier and a therapeutically effective amount of a compound of formula (I) as described above.
  • the present invention also provides methods for the treatment of HIV infection comprising administering to a host infected with HIV a pharmaceutically or
  • therapeutically effective amount it is meant an amount of a compound of the present invention effective to inhibit HIV infection or treat the symptoms of HIV infection in a host.
  • the compounds of formula (I) of the present invention are also useful for the inhibition of HIV in an ex vivo sample containing HIV or expected to be exposed to HIV.
  • the compounds of the present invention may be used to inhibit HIV present in a body fluid sample (for example, a serum or semen sample) which contains or is suspected to contain or be exposed to HIV.
  • the compounds provided by this invention are also useful as standard or reference compounds for use in tests or assays for determining the ability of an agent to inhibit viral replication and/or HIV protease, for example in a pharmaceutical research program.
  • the compounds of the present invention may be used as a control or reference compound in such assays and as a quality control standard.
  • the compounds of the present invention may be provided in a commercial kit or
  • the compounds of the present invention may also be useful as diagnostic reagents in diagnostic assays for the detection of HIV protease.
  • inhibition of the protease activity in an assay such as the assays described herein
  • a compound of the present invention would be indicative of the presence of HIV protease and HIV virus.
  • the present invention also provides a method of treating HIV infection in a mammal comprising
  • component (ii)) may provide a synergistic effect in inhibiting the replication of HIV.
  • the HIV inhibitory effects of the compound of formula (I) administered in combination with the RT inhibitor may be greater than the additive effect of each agent when administered alone.
  • the combination treatment using a compound of formula (I) in combination with a RT may provide a synergistic effect in inhibiting the replication of HIV.
  • the present invention provides an important advantage over current therapies for HIV.
  • component (i) and component (ii) of the present invention it is meant that the components are administered concurrently to the cell or mammal being treated.
  • each component may be administered at the same time or sequentially in any order at different points in time.
  • component (i) and component (ii) may be
  • This invention also includes pharmaceutical kits comprising or consisting essentially of a pharmaceutical composition comprising the cyclic HIV protease inhibitor compounds of formula (I) together with a pharmaceutical composition comprising an HIV RT inhibitor, and to methods of using such pharmaceutical kits for the inhibition of HIV and treatment of HIV infection.
  • This invention also includes combination products comprising pharmaceutical compositions comprising a cyclic HIV protease inhibitor compound of formula (I) in physical combination or in a single dosage unit with an HIV RT inhibitor, to pharmaceutical kits containing these combination products, and to methods of using these combination products for the inhibition of HIV and treatment of HIV infection.
  • HIV reverse transcriptase (RT) inhibitors useful in the method, combination products, and pharmaceutical kits of the present invention include, but are not limited to: nucleoside analogs, such as 3'-azido-3'-deoxythymidine (AZT, zidovidine), 2',3'-dideoxyinosine (DDI, ddI), 2',3'-dideoxycytidine (DDC, ddC), d4T, and 3TC; and non-nucleoside RT inhibitors, such as TIBO derivatives; BI-RG-587 and derivatives thereof;
  • nevirapine L-697,661 and derivatives thereof; LY 73497; Ro 18,893; L-743 , 726 and derivatives thereof; and
  • AZT is described by Chu et al., Tetrahedron Letters 29: 5349 (1988). AZT is available commercially as "Retrovir ® ", for which the product information, including dosage and administration, is given in Physicians' Desk Reference, 46th Edition, 1992, pp. 802-808.
  • asymmetrically substituted carbon atom may be isolated in optically active or racemic forms. It is well known in the art how to prepare optically active forms, such as by resolution of racemic forms or by synthesis, from optically active starting materials.
  • any variable for example, but not limited to, R 1 , R 5 , R 7 , R 9 , m, n etc.
  • its definition on each occurrence is independent of its definition at every other occurrence.
  • said group may optionally be
  • R 9 and R 9 at each occurrence is selected independently from the defined list of possible R 9 . Also, combinations of substituents and/or variables are permissible only if such combinations result in stable compounds.
  • alkyl is intended to include both branched and straight-chain saturated aliphatic
  • cycloalkyl represents an alkyl group of indicated number of carbon atoms attached through an oxygen bridge; "cycloalkyl” is intended to include saturated ring groups, including mono-,bi- or poly-cyclic ring systems, such as
  • bicyclopropyl cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, adamantyl and cyclooctyl
  • "biycloalkyl” is intended to include saturated bicyclic ring groups such as [3.3.0]bicyclooctane, [4.3.0]bicyclononane,
  • hydrocarbon chains of either a straight or branched configuration and one or more unsaturated carbon-carbon bonds which may occur in any stable point along the chain such as ethenyl, propenyl, and the like
  • alkynyl is intended to include hydrocarbon chains of either a straight or branched configuration and one or more triple carbon-carbon bonds which may occur in any stable point along the chain, such as ethynyl, propynyl and the like.
  • Alkylcarbonyl is intended to include an alkyl group of an indicated number of carbon atoms attached through a carbonyl group to the residue of the compound at the designated location.
  • Alkylcarbonylamino is intended to include an alkyl group of an indicated number of carbon atoms attached through a carbonyl group to an amino bridge, where the bridge is attached to the residue of the compound at the designated location.
  • Alkylcarbonyloxy is intended to include an alkyl group of an indicated number of carbon atoms attached to a carbonyl group, where the carbonyl group is attached through an oxygen atom to the residue of the compound at the designated location.
  • Halo or "halogen” as used herein refers to fluoro, chloro, bromo, and iodo; and "counterion” is used to represent a small, negatively charged species such as chloride, bromide, hydroxide, acetate, sulfate, and the like.
  • aryl or “aromatic residue” is intended to mean phenyl or naphthyl; the term
  • arylalkyl represents an aryl group attached through an alkyl bridge.
  • C 7 -C 10 arylalkyl is intended to refer to an aryl group
  • (C 1 -C 3 alkyl)aryl is intended to refer to a C 1 -C 3 alkyl group which is attached through an aryl ring to the residue of the indicated compound
  • aryl(C 1 -C 3 alkyl) is intended to refer to an aryl group attached through a C 1 -C 3 alkyl group to the residue of the indicated compound.
  • carbocycle or “carbocyclic residue” is intended to mean any stable 3- to 7-membered monocyclic or bicyclic or 7 - to 14-membered bicyclic or tricyclic or an up to 26-membered polycyclic carbon ring, any of which may be saturated, partially unsaturated, or aromatic.
  • carbocyles include, but are not limited to, cyclopropyl,
  • cyclopentyl cyclohexyl, phenyl, biphenyl, naphthyl, indanyl, adamantyl, or tetrahydronaphthyl (tetralin).
  • heterocycle is intended to mean a stable 5- to 7- membered monocyclic or
  • bicyclic or 7 - to 10-membered bicyclic heterocyclic ring which is either saturated or unsaturated, and which consists of carbon atoms and from 1 to 4 heteroatoms independently selected from the group consisting of N, O and S and wherein the nitrogen and sulfur heteroatoms may optionally be oxidized, and the nitrogen may
  • heterocyclic ring may be attached to its pendant group at any heteroatom or carbon atom which results in a stable structure.
  • the heterocyclic rings described herein may be substituted on carbon or on a nitrogen atom if the resulting
  • heterocycles include, but are not limited to, pyridinyl, pyrimidinyl, furanyl, thienyl, pyrrolyl, pyrazolyl, imidazolyl, tetrazolyl, benzofuranyl, benzothiophenyl, indolyl, indolenyl, quinolinyl, isoquinolinyl, benzimidazolyl, piperidinyl, 4-piperidonyl, pyrrolidinyl, 2-pyrrolidonyl, pyrrolinyl, tetrahydrofuranyl,
  • tetrahydroquinolinyl tetrahydroisoquinolinyl, decahydroquinolinyl or octahydroisoquinolinyl, azocinyl, triazinyl, 6H-1,2,5-thiadiazinyl, 2H, 6H- 1 , 5 , 2 -dithiazinyl, thiophenyl, thianthrenyl, pyranyl,
  • phenothiazinyl furazanyl, phenoxazinyl, isochromanyl, chromanyl, imidazolidinyl, imidazolinyl, pyrazolidinyl, pyrazolinyl, piperazinyl, indolinyl, isoindolinyl, quinuclidinyl, morpholinyl, oxazolidinyl,
  • heterocycles include, but are not limited to, pyridinyl, furanyl, thienyl,
  • benzimidazolyl 1H-indazolyl, oxazolidinyl, thiazolyl, 1,3,4-thiadiazolyl, oxazolyl, 1,3,4-oxadiazolyl, benzothiazolyl, benzotriazolyl, benzisoxazolyl, or benzoxazolinyl.
  • substituent may be bonded to any atom on the ring.
  • substituent may be listed without indicating the atom via which such substituent is bonded to the rest of the compound of formula I, then such substituent may be bonded via any atom in such substituent.
  • substituent is piperazinyl, piperidinyl, or tetrazolyl, unless specified otherwise, said
  • piperazinyl, piperidinyl, tetrazolyl group may be bonded to the rest of the compound of formula (I) via any atom in such piperazinyl, piperidinyl, tetrazolyl group.
  • stable compound or stable structure it is meant herein a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture, and formulation into an efficacious therapeutic agent.
  • substituted means that an one or more hydrogen on the designated atom is replaced with a selection from the indicated group, provided that the designated atom's normal valency is not exceeded, and that the substitution results in a stable compound.
  • 2 hydrogens on the atom are replaced.
  • amine protecting group means any group known in the art of organic synthesis for the protection of amine groups. Such amine
  • protecting groups include, but are not limited to, the following: 1) acyl types such as formyl,
  • hydroxy protecting group refers to any group known in the art of organic synthesis for the protection of hydroxyl groups.
  • hydroxy protecting group reagent refers to any reagent known in the art of organic synthesis for the protection of hydroxy groups which may be reacted with an hydroxy to provide an hydroxy group protected with an hydroxy protecting group.
  • protecting groups include those listed in Greene and Wuts, "Protective Groups in Organic
  • the hydroxy protecting groups are base-stable and can include, but are not limited to acyl types, aromatic carbamate types and alkyl types. Exemplary are methyl, methoxymethyl (MOM) , methylthiomethyl, benzyloxymethyl, t-butoxymethyl, 2-methoxyethoxymethyl, 2,2,2-trichloroethoxymethyl, 2-(trimethylsilyl)ethoxymethyl (SEM), tetrahydropyranyl, tetrahydrofuranyl, t-butyl, triphenylmethyl, trimethylsilyl, triethylsilyl,
  • Hydroxy protecting groups also include diol
  • cyclic acetal protecting group includes any protecting group known in the art of organic synthesis for the protection of 1,2-diol group through formation of a cyclic acetal or cyclic ketal group.
  • protecting groups include, but are not limited to, those listed in Greene and Wuts, "Protective Groups in Organic Synthesis", John Wiley & Sons, New York (1991), the disclosure of which is hereby
  • cyclic acetal 1,2-diol protecting groups are methylene acetal, ethylidene acetal, 2,2,2-trichloroethylidene acetal, acetonide, cycloheptylidene ketal, cyclopentylidene ketal, cyclohexylidene ketal, benzylidene acetal, phenanthrylidene, and methoxymethylene acetal.
  • Such ketal rings or ketal protecting groups are well known in the art of organic synthesis and typically include, for example, substituted or unsubstituted carbocyclic diethers, dithioethers, mixed ethers, enol ethers or ketones.
  • peptide as used herein means a compound that consists of two or more amino acids (as defined herein) that are linked by means of a peptide bond.
  • peptide also includes compounds containing both peptide and non-peptide components, such as
  • pseudopeptide or peptide mimetic residues or other non-amino acid components Such a compound containing both peptide and non-peptide components may also be referred to as a "peptide analog”.
  • peptide bond means a covalent amide linkage formed by loss of a molecule of water between the carboxyl group of one amino acid and the amino group of a second amino acid.
  • pharmaceutically acceptable salts refer to derivatives of the disclosed compounds wherein the parent compound of formula (I) is modified by making acid or base salts of the compound of formula (I).
  • Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines; alkali or organic salts of acidic residues such as carboxylic acids; and the like.
  • Prodrugs are considered to be any covalently bonded carriers which release the active parent drug according to formula (I) in vivo when such prodrug is administered to a mammalian subject.
  • Prodrugs of the compounds of formula (I) are prepared by modifying functional groups present in the compounds in such a way that the modifications are cleaved, either in routine manipulation or in vivo, to the parent compounds.
  • Prodrugs include compounds of formula (I) wherein hydroxy, amine, or sulfhydryl groups are bonded to any group that, when administered to a mammalian subject, cleaves to form a free hydroxyl, amino, or sulfhydryl group, respectively.
  • Examples of prodrugs include, but are not limited to, acetate, formate, or benzoate derivatives of alcohol and amine functional groups in the compounds of formula (I); phosphate esters,
  • the pharmaceutically acceptable salts of the compounds of formula (I) include the conventional non-toxic salts or the quaternary ammonium salts of the compounds of formula (I) formed, for example, from non-toxic inorganic or organic acids.
  • such conventional non-toxic salts include those derived from inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric and the like; and the salts prepared from organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic, maleic,
  • salicylic sulfanilic, 2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isethionic, and the like.
  • the pharmaceutically acceptable salts of the present invention can be synthesized from the compounds of formula (I) which contain a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent, or in a mixture of the two; generally,
  • nonaqueous media like ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred.
  • Lists of suitable salts are found in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, PA, 1985, p. 1418, the disclosure of which is hereby
  • the compounds of the present invention can be prepared in a number of ways well known to one skilled in the art of organic synthesis.
  • the compounds of the present invention can be synthesized using the methods described below, together with synthetic methods known in the art of synthetic organic chemistry, or variations thereon as appreciated by those skilled in the art.
  • Preferred methods include but are not limited to those methods described below.
  • the compounds of the present invention may be synthesized using the general synthetic procedures described below. Each of the references cited below are hereby incorporated herein by reference.
  • the compounds of the present invention are prepared by modifications of the procedures disclosed for the synthesis of cyclic urea HIV protease inhibitors in PCT Patent Applications WO 93/07128 published April 15, 1993 and WO 94/19329 published September 1, 1994, and in copending commonly assigned U.S. Patent Application USSN 08/197,630 filed February 16, 1994. The disclosure of these references is incorporated herein by reference .
  • R 1 can be -X(CH 2 ) n COOH or as defined above for compounds of formula I; R 3 , R 4 and X are as defined above for compounds of formula I; and R is a hydroxyl or diol protecting group, by condensation with
  • diisopropylcarbodiimide or 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC)) method
  • EDC 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride
  • active ester p-nitrophenyl ester, N-hydroxysuccinic imido ester
  • carbonyldiimidazole method or phosphorus reagents such as BOP-Cl.
  • the desired amides can be prepared by conversion of acid (IIIc) to the corresponding acid chloride and treatment of the acyl chloride with a 2-aminoheteroaryl in the presence of a base.
  • carboxylic acids to acid chlorides are described in (March, Adv. Org. Chem. 1985, p. 1146, J. Wiley & Son, USA) and include, for example, treatment of the acid with oxalyl chloride in the presence of a catalytic amount of N,N'-dimethylformamide.
  • R 1 can be -X(CH 2 ) n Q or as defined above for compounds of formula I; R 3 , R 4 and X are as defined above for compounds of formula I; R is a hydroxyl or diol protecting group, and Q is -OH or -NH 2 , with a bromoheteroaryl in the presence of a strong base, such as sodium hydride as shown in Scheme 3.
  • a strong base such as sodium hydride as shown in Scheme 3.
  • Trisilylated thiophene XIV and disilylated thiophene XIII To a stirred, cooled (-78°C) solution of 1.60 g(2.86 mmol) of thiophene I in 10 mL of THF was added 2.6 mL(6.5 mmol) of a 2.5M solution of n-BuLi in hexanes. The solution was stirred 15 min. warming to 0°C and then re-cooled to -78°C. The solution was treated with 1.27 mL(9.0 mmol) of trimethylsilylisothiocyanate, and stirring was continued for 30 min while the solution warmed to 0°C.
  • the crude acid was dissolved in 5 mL of THF and treated with 2 mL of 1M tetra-(n-butylammonium fluoride) (TBAF) in THF. The solution was stirred 16 h and poured into water. The mixture was extracted with EtOAc, and the organic extract was washed with brine, dried (MgSO 4 ), and concentrated under reduced pressure to give an oil.
  • TBAF tetra-(n-butylammonium fluoride)
  • the crude product was dissolved in 2 mL of methanol and treated with 0.1 mL of con. aq. HCl. The solution was stirred 16 h, poured into water, and extracted with ethyl acetate. The organic extract was dried (MgSO 4 ) and concentrated under reduced pressure to afford a glass.
  • the crude product was chromatographed on silica
  • the compounds of this invention possess retroviral protease inhibitory activity, in particular, HIV
  • the compounds of formula (I) possess HIV protease inhibitory activity and are therefore useful as antiviral agents for the treatment of HIV infection and associated diseases.
  • the compounds of formula (I) possess HIV protease inhibitory activity and are effective as inhibitors of HIV growth.
  • the protease inhibitory activity of the compounds of the present invention is demonstrated using standard assays of protease activity, for example, using the assay described below for assaying inhibitors of HIV protease activity.
  • the ability of the compounds of the present invention to inhibit viral growth or infectivity is demonstrated in standard assay of viral growth or infectivity, for example, using the assays described below.
  • the compounds of the present invention can be demonstrated to inhibit HIV-protease activity in vivo using the HIV-1 Protease Transgenic Mouse animal model described in PCT Application Publication Number
  • the compounds of the present invention can also be demonstrated to exhibit in vivo HIV inhibitory activity using the HIV-1/CEM Mouse Xenotransplant animal model described in PCT Application Publication Number WO 94/19329.
  • the compounds of the present invention inhibit HIV growth and infectivity, they may be used as HIV antivirals for the inhibition of HIV ex vivo in a biological sample which contains HIV or is suspected to contain HIV or to be exposed to HIV.
  • compounds of the present invention can be used to inhibit HIV present in body fluid samples, for example, a serum or semen sample which contains, or is suspected of containing, HIV.
  • the samples can be treated, for example, in a method similar to those described below for the HIV RNA Assay or the HIV Yield Reduction Cell Assay.
  • the compounds of the present invention can be utilized as standard or reference compounds for use in procedures for the screening for agents which inhibit viral replication and/or HIV protease.
  • the compounds according to the present invention can be used as positive controls of inhibitory activity in such assays and as a quality control standard.
  • the assays can be performed, for example, as described below under the HIV Protease Inhibition Assay, the HIV RNA Assay, or the HIV Yield Reduction Cell Assay.
  • the compounds of the present invention can be provided in a commercial kit or container comprising a compound of this
  • the compounds of the present invention exhibit selectivity for HIV protease
  • the compounds of the present claims can also be used as diagnostic reagents in diagnostic assays for the
  • ⁇ g denotes microgram
  • mg denotes milligram
  • g denotes gram
  • ⁇ L microliter
  • mL denotes milliliter
  • L denotes liter
  • nM denotes nanomolar
  • ⁇ M denotes micromolar
  • mM denotes
  • Sigma stands for the Sigma-Aldrich Corp. of St. Louis, MO. A compound is considered to be active if it has an IC 50 or K i value of less than about 1 mM for the
  • HIV protease with a K i value of ⁇ 1 nM and inhibit virus with an IC 90 value of ⁇ 0.05 ⁇ g/mL.
  • Protease Inclusion bodies of E. coli harboring plasmid containing plasmid T1718R with a synthetic gene coding for a single-chain tethered dimer of HIV protease were prepared as described in Cheng et al. (Proc. Natl. Acad. Sci. USA, 87, 9660-9664, 1990). Active protease was prepared as described therein by extraction with 67% acetic acid, dilution 33-fold with water, dialysis against water and then against a refolding buffer consisting of 20 mM MES, 1 mM dithiothreitol and 10% glycerol. Protease was stored as a stock preparation at 10 ⁇ M in refolding buffer.
  • Substrate Peptide of sequence: aminobenzoyl-Ala-Thr-His-Gln-Val-Tyr-Phe(NO 2 )-Val-Arg-Lys-Ala containing p-nitrophenylalanine, was prepared by solid phase synthesis as previously described Cheng et al. (Proc. Natl. Acad. Sci. USA, 87, 9660-9664, 1990). Stock solutions of 2 mM substrate were prepared in DMSO.
  • Inhibitory compounds were dissolved in sufficient DMSO to make 3 mM stock solutions. All further dilutions were prepared in "assay buffer": 1 M NaCl, 50 mM MES, pH 5.5, 1 mM EDTA, 1mM DTT, 20% glycerol.
  • Enzyme reaction In a 2 mL screw-cap centrifuge tube were added 50 ⁇ l protease (final concentration 0.25 nM) and 0.1 mL inhibitory compound (final concentration 0.1-12,500). After 15 min preincubation at room temperature
  • HPLC measurement of product formation The product (aminobenzoyl -Ala- Thr-His -Gln-Val - Tyr) was separated from substrate on a Pharmacia MonoQ anion exchange column. The injection volume was 0.2 mL. The mobile phases were A (20 mM trisHCl, pH 9.0, 0.02%
  • Plasmid pDAB 72 containing both gag and pol sequences of BH10 (bp 113-1816) cloned into PTZ 19R was prepared according to Erickson-Viitanen et al., AIDS Research and Human Retroviruses (1989) 5: 577. The plasmid was linearized with Bam HI prior to the
  • RNA transcripts were dissolved in water, and stored at -70°C. The concentration of RNA was determined from the A 260 .
  • Biotinylated capture probes were purified by HPLC after synthesis on an Applied Biosystems (Foster City, CA) DNA synthesizer by addition of biotin to the 5' terminal end of the oligonucleotide, using the biotin-phosphoramidite reagent of Cocuzza, Tet. Lett. (1989)
  • gag biotinylated capture probe (5-biotin-CTAGCTCCCTGCTTGCCCATACTA 3') was complementary to nucleotides 889-912 of HXB2 and the pol biotinylated capture probe (5'-biotin -CCCTATCATTTTTGGTTTCCAT 3' ) was complementary to nucleotides 2374-2395 of HXB2.
  • Alkaline phosphatase conjugated oligonucleotides used as reporter probes were prepared by Syngene (San Diego, CA.).
  • the pol reporter probe was prepared by Syngene (San Diego, CA.). The pol reporter probe
  • the reporter probes were prepared as 0.5 ⁇ M stocks in 2 x SSC (0.3 M NaCl, 0.03 M sodium citrate), 0.05 M Tris pH 8.8, 1 mg/mL BSA.
  • the biotinylated capture probes were prepared as 100 ⁇ M stocks in water.
  • Nunc-immunomodule microtiter plate strips were coated by addition of 200 ⁇ L of streptavidin (30 ⁇ g/mL, Scripps, La Jolla, CA) in freshly prepared 10 mM sodium carbonate (pH 9.6). Plates were incubated overnight at 4°C. Streptavidin solution was aspirated from the wells and a blocking buffer composed of phosphate buffered saline (PBS), 20 mg/mL bovine serum albumin
  • MT-2, CEM, and H9 cells were maintained in RPMI 1640 supplemented with 5% fetal calf serum (FCS), 2 mM L-glutamine and 50 ⁇ g/mL gentamycin, all from Gibco.
  • FCS fetal calf serum
  • HIV-1 Laboratory strains of HIV-1 (RF, MN and IIIB) were propagated in H9 cells in the same medium. Virus stocks were prepared approximately 1 month after acute
  • Infectious titers of HIV-l(RF) stocks were 1-3 ⁇ 10 7 PFU (plaque forming units) /mL as measured by plaque assay on MT-2 cells (see below). Each aliquot of virus stock used for infection was thawed only once. In some cases, infected H9 cells were shifted to Dulbecco's modified Eagle's medium 3-10 days before collection of virus in order to generate virus stocks in medium with low biotin content. Clinical isolates of HIV that had been
  • MT-2 cells passaged once in MT-2 cells were used to infect fresh MT-2 cells in RPMI medium. Three days after infection, cells were pelleted, resuspended and culture continued in Dulbecco's modified Eagle's medium as above. Virus stocks of clinical isolates were prepared 10-15 days after infection when cytopathic effects were apparent in the culture. For evaluation of antiviral efficacy, cells to be infected were subcultured one day prior to infection.
  • Virus was added and culture continued for 3 days at 37°C. In some experiments, virus was removed after an initial adsorption period.
  • HIV-1 infected cells were pelleted by
  • Hybridization was carried out in sealed microfuge tubes or in sealed U bottom 96 well tissue culture plates ( ⁇ unc or Costar) for 16-20 hours at 37°C.
  • hybridization reactions were diluted three-fold with deionized water to a final guanidinium isothiocyanate concentration of 1 M and aliquots (150 ⁇ L) were
  • immobilized complex of capture probe and hybridized target RNA was carried out in the washed streptavidin coated well by addition of 120 ⁇ l of a hybridization cocktail containing 4 X SSC, 0.66% Triton X 100, 6.66% deionized formamide, 1 mg/mL BSA and 5 nM reporter probe. After hybridization for one hour at 37°C, the plate was again washed 6 times. Immobilized alkaline phosphatase activity was detected by addition of 100 ⁇ L of 0.2 mM 4-methylumbelliferyl phosphate (MUBP, JBL Scientific) in buffer ⁇ (2.5 M diethanolamine pH 8.9 (JBL
  • MT-2 cells 50 ⁇ L were added to a final concentration of 5 ⁇ 10 5 per mL (1 ⁇ 10 5 per well). Cells were incubated with compounds for 30 minutes at 37°C in a CO 2 incubator.
  • an appropriate dilution of HIV-1 (RF) virus stock 50 ⁇ L was added to culture wells
  • test compounds containing cells and dilutions of the test compounds.
  • the final volume in each well was 200 ⁇ L.
  • Eight wells per plate were left uninfected with 50 ⁇ L of medium added in place of virus, while eight wells were infected in the absence of any antiviral compound.
  • parallel plates were cultured without virus infection.
  • HIV RNA was quantitated as described above.
  • a standard curve prepared by adding known amounts of pDAB 72 in vi tro RNA transcript to wells containing lysed uninfected cells, was run on each microtiter plate in order to determine the amount of viral RNA made during the infection.
  • IC 90 value concentration of compound required to reduce the HIV RNA level by 90%
  • ddC dideoxycytidine
  • IC 90 values of other antiviral compounds, both more and less potent than ddC were reproducible using several stocks of HIV-1 (RF) when this procedure was followed.
  • This concentration of virus corresponded to ⁇ 3 ⁇ 10 5 PFU (measured by plaque assay on MT-2 cells) per assay well and typically produced approximately 75% of the maximum viral RNA level achievable at any virus inoculum.
  • IC 90 values were determined from the percent reduction of net signal (signal from infected cell samples minus signal from uninfected cell samples) in the RNA assay relative to the net signal from infected, untreated cells on the same culture plate (average of eight wells). Valid performance of individual infection and RNA assay tests was judged according to three criteria. It was required that the virus infection should result in an RNA assay signal equal to or greater than the signal generated from 2 ng of pDAB 72 in vitro RNA transcript. The IC 90 for ddC, determined in each assay run, should be between 0.1 and 0.3 ⁇ g/mL. Finally, the plateau level of viral RNA produced by an effective protease inhibitor should be less than 10% of the level achieved in an uninhibited infection.
  • MT-2 a human T-cell line, was cultured in RPMI medium supplemented with 5% (v/v) heat
  • FCS fetal calf serum
  • RF gentamycin
  • Human immunodeficiency virus strains, HIV (3B) and HIV(RF) were propagated in H-9 cells in RPMI with 5% FCS.
  • Poly-L-lysine (Sigma) coated cell culture plates were prepared according to the method of Harada et al., Science (1985) 229: 563-566. MTT, 3-(4,5- dimethyl-thiazol-2yl)-2,5-diphenyltetrazolium bromide, was obtained from Sigma.
  • MT-2 cells (5 ⁇ 10 5 /mL) in 2.3 mL were mixed with 0.3 mL of the appropriate test compound solution and allowed to sit for 30 minutes at room temperature.
  • HIV (3B) or HIV (RF) ( ⁇ 5 ⁇ 10 5 plaque forming units/mL) in 0.375 mL was added to the cell and compound mixtures and incubated for one hour at 36°C. The mixtures were centrifuged at 1000 rpm for 10 minutes and the supernatants containing unattached virus were discarded. The cell pellets were suspended in fresh RPMI containing the appropriate concentrations of test compound and placed in a 36°C, 4% CO 2 incubator. Virus was allowed to replicate for 3 days. Cultures were centrifuged for 10 minutes at 1000 rpm and the
  • the virus titers of the progeny virus produced in the presence or absence of test compounds were:
  • Progeny virus suspensions were serially diluted in RPMI and 1.0 mL of each
  • dilution was added to 9 mL of MT-2 cells in RPMI.
  • Cells and virus were incubated for 3 hours at 36°C to allow for efficient attachment of the virus to cells.
  • Each virus and cell mixture was aliquoted equally to two wells of a six well poly-L-lysine coated culture plate and incubated overnight at 36°C, 4% CO 2 .
  • Liquid and unattached cells were removed prior to the addition of 1.5 mL of RPMI with 0.75% (w/v) Seaplaque agarose (FMC Corp.) and 5% FCS. Plates were incubated for 3 days and a second RPMI/agarose overlay was added.
  • test compounds were determined by the percent reduction in the virus titer with respect to virus grown in the absence of any inhibitors.
  • the tested compounds of the present invention exhibit HIV protease and HIV virus inhibitory activity in the assays described above.
  • the antiviral compounds of this invention can be administered as treatment for retroviral infections by any means that produces contact of the active agent with the agent's site of action, the retroviral protease, in the body of a mammal. They can be administered by any conventional means available for use in conjunction with pharmaceuticals, either as individual therapeutic agents or in a combination of therapeutic agents. They can be administered alone, but generally administered with a pharmaceutical carrier selected on the basis of the chosen route of administration and standard
  • the compounds of the present invention can be administered in oral dosage forms as tablets, capsules (each of which includes sustained release or timed release formulations), pills, powders, granules, elixirs, tinctures, suspensions, syrups, and emulsions.
  • the compounds of the present invention may also be administered in intravenous (bolus or infusion), intraperitoneal, subcutaneous, or intramuscular form, all using dosage forms well known to those of ordinary skill in the pharmaceutical arts.
  • An effective amount of the compound desired can be administered for the inhibition of HIV and the treatment of HIV infection.
  • the dosage administered will, of course, vary depending upon known factors, such as the
  • the daily oral dosage of each active ingredient when used for the indicated effects, will range between about 0.001 to 1000 mg/kg of body weight, preferably between about 0.01 to 100 mg/kg of body weight per day, and most preferably between about 1.0 to 20 mg/kg/day.
  • the most preferred doses may range from about 1 to about 10 mg/kg/minute during a constant rate infusion.
  • Compounds of the present invention may be administered in a single daily dose, or the total daily dosage may be
  • the compounds for the present invention may also be administered in intranasal form via topical use of suitable intranasal vehicles, or via transdermal routes, using those forms of transdermal skin patches well known to those of ordinary skill in that art.
  • the dosage administration will, of course, be continuous rather than intermittent throughout the dosage regimen.
  • the compounds herein described in detail can form the active ingredient, and are typically administered in admixture with suitable pharmaceutical diluents, excipients, or carriers (collectively referred to herein as a pharmaceutically acceptable carrier or carrier materials) suitably selected with respect to the intended form of administration, that is, oral tablets, capsules, elixirs, syrups and the like, and consistent with conventional pharmaceutical practices.
  • suitable pharmaceutical diluents, excipients, or carriers collectively referred to herein as a pharmaceutically acceptable carrier or carrier materials
  • suitable pharmaceutical diluents, excipients, or carriers suitably selected with respect to the intended form of administration, that is, oral tablets, capsules, elixirs, syrups and the like, and consistent with conventional pharmaceutical practices.
  • the active drug component can be combined with an oral, non-toxic, pharmaceutically acceptable, inert carrier such as lactose, starch, sucrose, glucose, methyl cellulose, magnesium stearate, dicalcium phosphate, calcium sulfate, mannitol, sorbitol and the like.
  • an oral, non-toxic, pharmaceutically acceptable, inert carrier such as lactose, starch, sucrose, glucose, methyl cellulose, magnesium stearate, dicalcium phosphate, calcium sulfate, mannitol, sorbitol and the like.
  • the oral drug components can be combined with any oral, non-toxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like.
  • suitable binders, lubricants, disintegrating agents, and coloring agents can also be incorporated into the mixture.
  • Suitable binders include starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth, or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes, and the like.
  • Lubricants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, and the like.
  • Disintegrators include, without limitation, starch, methyl cellulose, agar, bentonite, xanthan gum, and the like.
  • the compounds of the present invention can also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamallar vesicles, and multilamellar vesicles.
  • Liposomes can be formed from a variety of phospholipids, such as cholesterol, stearylamine, or phosphatidylcholines.
  • Compounds of the present invention may also be coupled with soluble polymers as targetable drug carriers.
  • soluble polymers can include
  • the compounds of the present invention may be coupled to a class of biodegradable polymers useful in achieving controlled release of a drug, for example, polylactic acid, polyglycolic acid, copolymers of polylactic and polyglycolic acid,
  • polyepsilon caprolactone polyhydroxy butyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacylates, and crosslinked or amphipathic block copolymers of hydrogels.
  • compositions suitable for injection compositions suitable for injection.
  • compositions contain from about 1 milligram to about 100 milligrams of active ingredient per dosage unit.
  • active ingredient will ordinarily be present in an amount of about 0.5-95% by weight based on the total weight of the dosage unit.
  • the active ingredient can be administered orally in solid dosage forms, such as capsules, tablets, and powders, or in liquid dosage forms, such as elixirs, syrups, and suspensions. It can also be administered parenterally, in sterile liquid dosage forms.
  • Gelatin capsules may contain the active ingredient and powdered carriers, such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, and the like. Similar diluents can be used to make compressed tablets. Both tablets and capsules can be manufactured as sustained release products to provide for continuous release of medication over a period of hours.
  • powdered carriers such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, and the like. Similar diluents can be used to make compressed tablets. Both tablets and capsules can be manufactured as sustained release products to provide for continuous release of medication over a period of hours.
  • Compressed tablets can be sugar coated or film coated to mask any unpleasant taste and protect the tablet from the atmosphere, or enteric coated for selective
  • Liquid dosage forms for oral administration can contain coloring and flavoring to increase patient acceptance.
  • water, a suitable oil, saline, aqueous dextrose (glucose), and related sugar solutions and glycols such as propylene glycol or polyethylene glycols are suitable carriers for parenteral solutions.
  • Solutions for parenteral administration preferably contain a water soluble salt of the active ingredient, suitable stabilizing agents, and if necessary, buffer substances.
  • suitable stabilizing agents such as sodium metabisulfite, sodium metabisulfite, sodium metabisulfite, sodium metabisulfite, sodium metabisulfite, sodium metabisulfite, sodium metabisulfite, sodium metabisulfite, sodium metabisulfite, sodium metabisulfite, sodium metabisulfite, sodium metabisulfite, sodium metabisulfite, sodium metabisulfite, sodium metabisulfite, sodium metabisulfite, sodium metabisulfite, sodium metabisulfite, sodium metabisulfite, sodium metabisulfite, sodium metabisulfite, sodium metabisulfite, sodium metabisulfite, sodium metabisulfite, sodium metabisulfite, sodium metabisulfite, sodium metabisulfite, sodium metabisulfite, sodium metabisulfite, sodium metabisulfite
  • bisulfite, sodium sulfite, or ascorbic acid are suitable stabilizing agents.
  • citric acid and its salts and sodium EDTA are also used.
  • parenteral solutions can contain
  • preservatives such as benzalkonium chloride, methyl- or propyl-paraben, and chlorobutanol.
  • Suitable pharmaceutical carriers are described in Remington's Pharmaceutical Sciences, Mack Publishing
  • a large number of unit capsules are prepared by filling standard two-piece hard gelatin capsules each with 100 milligrams of powdered active ingredient, 150 milligrams of lactose, 50 milligrams of cellulose, and 6 milligrams magnesium stearate.
  • a mixture of active ingredient in a digestable oil such as soybean oil, cottonseed oil or olive oil is prepared and injected by means of a positive
  • the dosage unit was 100 milligrams of active ingredient, 0.2 milligrams of colloidal silicon dioxide, 5 milligrams of magnesium stearate, 275 milligrams of microcrystalline cellulose,
  • administration by injection is prepared by stirring 1.5% by weight of active ingredient in 10% by volume
  • propylene glycol and water.
  • the solution is made isotonic with sodium chloride and sterilized.
  • An aqueous suspension is prepared for oral
  • each 5 mL contain 100 mg of finely divided active ingredient, 200 mg of sodium carboxymethyl cellulose, 5 mg of sodium benzoate, 1.0 g of sorbitol solution, U.S.P., and 0.025 mL of vanillin.
  • the compounds of formula (I) of the present invention may be administered in combination with a second therapeutic agent, such as a second HIV
  • the compound of formula (I) and such second therapeutic agent can be administered separately or as a physical combination in a single dosage unit, in any dosage form and by various routes of administration, as described above.
  • the compound of formula (I) may be formulated together with the second therapeutic agent in a single dosage unit (that is, combined together, for example, in one capsule, tablet, powder, or liquid).
  • the compound of formula (I) and the second therapeutic agent may be administered essentially at the same time, or in any order; for example, the compound of formula (I) may be administered first, followed by administration of the second agent.
  • the present invention also includes pharmaceutical kits useful, for example, for the treatment of HIV infection, which comprise one or more containers
  • kits may further include, if desired, one or more of various conventional pharmaceutical kit components, such as, for example, containers with one or more pharmaceutically acceptable carriers, additional containers, etc., as will be readily apparent to those skilled in the art.
  • Printed instructions either as inserts or as labels, indicating quantities of the components to be administered, guidelines for
  • administration, and/or guidelines for mixing the components may also be included in the kit.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

Urées cycliques substituées et dérivés de ces urées renfermant des composés de la formule (II). Ces composés se révèlent utiles comme inhibiteurs de la protéase de VIH. L'invention porte également sur des compositions pharmaceutiques renfermant de tels composés et sur des procédés d'utilisation de ces composés dans le traitement de l'infection par VIH. L'invention porte également sur l'utilisation de ces composés dans des procédés d'identification d'inhibiteurs de la protéase de VIH et pour l'inhibition ou la détection du VIH dans un échantillon de liquide organique.
PCT/US1996/003426 1995-03-17 1996-03-13 Urees cycliques inhibitrices de la protease de vih WO1996029329A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
AU53100/96A AU5310096A (en) 1995-03-17 1996-03-13 Cyclic urea hiv protease inhibitors
EP96909680A EP0815108A1 (fr) 1995-03-17 1996-03-13 Urees cycliques inhibitrices de la protease de vih

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US40624095A 1995-03-17 1995-03-17
US08/406,240 1995-03-17
US08/613,554 1996-03-11
US08/613,554 US5683999A (en) 1995-03-17 1996-03-11 Cyclic urea HIV protease inhibitors

Publications (1)

Publication Number Publication Date
WO1996029329A1 true WO1996029329A1 (fr) 1996-09-26

Family

ID=27019433

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1996/003426 WO1996029329A1 (fr) 1995-03-17 1996-03-13 Urees cycliques inhibitrices de la protease de vih

Country Status (4)

Country Link
EP (1) EP0815108A1 (fr)
AU (1) AU5310096A (fr)
CA (1) CA2215536A1 (fr)
WO (1) WO1996029329A1 (fr)

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5932570A (en) * 1996-11-08 1999-08-03 Dupont Pharmaceuticals Company 1-(3-aminoindazol-5-yl)-3-phenylmethyl-cyclic ureas useful as HIV protease inhibitors
US5985867A (en) * 1997-03-31 1999-11-16 Dupont Pharmaceuticals Company Indazoles of cyclic ureas useful as HIV protease inhibitors
WO2000073284A3 (fr) * 1999-05-26 2001-04-05 Du Pont Pharm Co Inhibiteurs de transcriptase inverse du vih a base de 1,4-benzodiazepin-2-ones
US6218386B1 (en) 1996-11-08 2001-04-17 Dupont Pharmaceuticals A1-(3-aminoindazol-5-yl)-3 butyl-cyclic urea useful as a HIV protease inhibitor
FR2810039A1 (fr) * 2000-06-13 2001-12-14 Centre Nat Rech Scient Composes urees cycliques et leur procede de preparation
US7777030B2 (en) 2005-12-29 2010-08-17 Centre National de la Recherge Scientifique (CNRS) Compositions and methods for the treatment and prevention of disease
US8865180B2 (en) 2006-04-10 2014-10-21 Infectious Disease Research Institute Compounds and methods for diagnosis and treatment of leishmaniasis
JP2016508505A (ja) * 2013-02-07 2016-03-22 メルク パテント ゲゼルシャフト ミット ベシュレンクテル ハフツングＭｅｒｃｋ Ｐａｔｅｎｔ Ｇｅｓｅｌｌｓｃｈａｆｔ ｍｉｔ ｂｅｓｃｈｒａｅｎｋｔｅｒ Ｈａｆｔｕｎｇ ピリダジノン−アミド誘導体

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993007128A1 (fr) * 1991-10-11 1993-04-15 The Du Pont Merck Pharmaceutical Company Urees cycliques et analogues utiles en tant qu'inhibiteurs de la protease retrovirale
WO1994019329A1 (fr) * 1993-02-26 1994-09-01 The Du Pont Merck Pharmaceutical Company Carbonyles cycliques substitues et leurs derives utiles comme inhibiteurs de protease retrovirale

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993007128A1 (fr) * 1991-10-11 1993-04-15 The Du Pont Merck Pharmaceutical Company Urees cycliques et analogues utiles en tant qu'inhibiteurs de la protease retrovirale
WO1994019329A1 (fr) * 1993-02-26 1994-09-01 The Du Pont Merck Pharmaceutical Company Carbonyles cycliques substitues et leurs derives utiles comme inhibiteurs de protease retrovirale

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
P.Y.S. LAM ET AL.: "Rational design of potent, bioavailable, nonpeptide cyclic ureas as HIV protease inhibitors", SCIENCE, vol. 263, 21 January 1994 (1994-01-21), WASHINGTON DC US, pages 380 - 4, XP002006572 *

Cited By (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6451999B2 (en) 1996-11-08 2002-09-17 Bristol-Myers Squibb Pharma Company 1-(3-aminoindazol-5-yl)-3-butyl-cyclic urea useful as a HIV protease inhibitor
US6218386B1 (en) 1996-11-08 2001-04-17 Dupont Pharmaceuticals A1-(3-aminoindazol-5-yl)-3 butyl-cyclic urea useful as a HIV protease inhibitor
US5932570A (en) * 1996-11-08 1999-08-03 Dupont Pharmaceuticals Company 1-(3-aminoindazol-5-yl)-3-phenylmethyl-cyclic ureas useful as HIV protease inhibitors
US5985867A (en) * 1997-03-31 1999-11-16 Dupont Pharmaceuticals Company Indazoles of cyclic ureas useful as HIV protease inhibitors
WO2000073284A3 (fr) * 1999-05-26 2001-04-05 Du Pont Pharm Co Inhibiteurs de transcriptase inverse du vih a base de 1,4-benzodiazepin-2-ones
US6462037B1 (en) 1999-05-26 2002-10-08 Bristol-Myers Squibb Pharma Company 1,4-benzodiazepin-2-ones useful as HIV reverse transcriptase inhibitors
FR2810039A1 (fr) * 2000-06-13 2001-12-14 Centre Nat Rech Scient Composes urees cycliques et leur procede de preparation
US7186828B2 (en) 2000-06-13 2007-03-06 Immupharma (France) Sa Cyclic urea compounds and preparation thereof
WO2001096318A1 (fr) * 2000-06-13 2001-12-20 Centre National De La Recherche Scientifique Composes urees cycliques et leur procede de preparation
EP1640368A2 (fr) 2000-06-13 2006-03-29 The Centre National de la Recherche Scientifique Composés urées cycliques et leur procédé de préparation
JP2004503546A (ja) * 2000-06-13 2004-02-05 サントル・ナショナル・ドゥ・ラ・ルシェルシュ・シャンティフィク 環式尿素化合物及びその調製方法
EP1640368A3 (fr) * 2000-06-13 2007-03-28 The Centre National de la Recherche Scientifique Composés urées cycliques et leur procédé de préparation
US7777030B2 (en) 2005-12-29 2010-08-17 Centre National de la Recherge Scientifique (CNRS) Compositions and methods for the treatment and prevention of disease
US8865180B2 (en) 2006-04-10 2014-10-21 Infectious Disease Research Institute Compounds and methods for diagnosis and treatment of leishmaniasis
JP2016508505A (ja) * 2013-02-07 2016-03-22 メルク パテント ゲゼルシャフト ミット ベシュレンクテル ハフツングＭｅｒｃｋ Ｐａｔｅｎｔ Ｇｅｓｅｌｌｓｃｈａｆｔ ｍｉｔ ｂｅｓｃｈｒａｅｎｋｔｅｒ Ｈａｆｔｕｎｇ ピリダジノン−アミド誘導体

Also Published As

Publication number Publication date
AU5310096A (en) 1996-10-08
CA2215536A1 (fr) 1996-09-26
EP0815108A1 (fr) 1998-01-07

Similar Documents

Publication Publication Date Title
EP0686151B1 (fr) Carbonyles cycliques substitues et leurs derives utiles comme inhibiteurs de protease retrovirale
US5683999A (en) Cyclic urea HIV protease inhibitors
WO1997008150A1 (fr) Urees cycliques substituees et leurs derives utiles en tant qu'inhibiteurs de protease retrovirale
WO1996029329A1 (fr) Urees cycliques inhibitrices de la protease de vih
US7186830B2 (en) Tricyclic 2-pyrimidone compounds useful as HIV reverse transcriptase inhibitors
WO2000000478A1 (fr) Quinoxalin-2(1h)-ones substitues utilises comme inhibiteurs de transcriptase inverse du vih
JPH11506765A (ja) Ｎ，ｎ´−ジ置換環状尿素の調製方法
US5877170A (en) Substitute caprolactams and derivatives thereof useful for treatment of HIV disease
US5763469A (en) Substituted cyclic ureas and derivatives thereof useful as retroviral protease inhibitors
US6204262B1 (en) 1,3-Benzodiazepin-2-ones and 1,3-Benzoxazepin-2-ones useful as HIV reverse transcriptase inhibitors
US5932570A (en) 1-(3-aminoindazol-5-yl)-3-phenylmethyl-cyclic ureas useful as HIV protease inhibitors
US5543517A (en) Substituted bicyclic phosphoramides and derivatives thereof
US6313110B1 (en) Substituted 2H-1,3-diazapin-2-one useful as an HIV protease inhibitor
EP0937067B1 (fr) Urees 1-(3-aminoindazol-5-yl)-3-phenylmethyl-cycliques utiles en tant qu'inhibiteurs de protease de vih
US7015214B2 (en) Cyanamide, alkoxyamino, and urea derivatives of 1,3-benzodiazepine as HIV reverse transcriptase inhibitors
US20010011088A1 (en) 1-(3-aminoindazol-5-yl)-3-butyl-cyclic urea useful as a HIV protease inhibitor
AU703962B2 (en) Substituted cyclic carbonyls and derivatives
KR20000053123A (ko) (4ｒ,5ｓ,6ｓ,7ｒ)-헥사히드로-1-[5-(3-아미노인다졸)메틸]-3-부틸-5,6-디히드록시-4,7-비스[페닐메틸]-2ｈ-1,3-디아제핀-2-온,이의 제조 방법 및 ｈｉｖ 프로테아제 억제제로서의 용도
HK1015767A (en) Substituted cyclic carbonyls and derivatives thereof useful as retroviral protease inhibitors
CA2156594A1 (fr) Carbonyles cycliques substitues et leurs derives, utilises comme inhibiteurs de la protease retrovirale
JP2003534230A (ja) Ｈｉｖ逆転写酵素阻害剤として有用な１，３−ベンゾジアゼピン−２−オンおよび１，３−ベンゾオキサゼピン−２−オン
AU2003203047A1 (en) Tricyclic 2-pyrimidone compounds useful as HIV reverse transcriptase inhibitors

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AU BR CA CN CZ EE HU JP KR LT LV MX NO NZ PL RO SG SI SK UA VN AM AZ BY KG KZ MD RU TJ TM

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FI FR GB GR IE IT LU MC NL PT SE

DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
121 Ep: the epo has been informed by wipo that ep was designated in this application
ENP Entry into the national phase

Ref document number: 2215536

Country of ref document: CA

Ref country code: CA

Ref document number: 2215536

Kind code of ref document: A

Format of ref document f/p: F

WWE Wipo information: entry into national phase

Ref document number: 1996909680

Country of ref document: EP

WWP Wipo information: published in national office

Ref document number: 1996909680

Country of ref document: EP

WWR Wipo information: refused in national office

Ref document number: 1996909680

Country of ref document: EP

WWW Wipo information: withdrawn in national office

Ref document number: 1996909680

Country of ref document: EP