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WO1995006061A1 - Inhibiteurs de proteases retrovirales et leurs combinaisons - Google Patents

Inhibiteurs de proteases retrovirales et leurs combinaisons Download PDF

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Publication number
WO1995006061A1
WO1995006061A1 PCT/US1994/008697 US9408697W WO9506061A1 WO 1995006061 A1 WO1995006061 A1 WO 1995006061A1 US 9408697 W US9408697 W US 9408697W WO 9506061 A1 WO9506061 A1 WO 9506061A1
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WO
WIPO (PCT)
Prior art keywords
amino
tert
butanol
phenyl
butylcarbamoyl
Prior art date
Application number
PCT/US1994/008697
Other languages
English (en)
Inventor
Min S. Chang
James C. Stolzenbach
John J. Talley
Michael L. Vazquez
Daniel P. Getman
Richard A. Mueller
James C. Ottinger
Gary A. Decrescenzo
Original Assignee
G.D. Searle & Co.
Monsanto 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 US08/253,531 external-priority patent/US5750648A/en
Application filed by G.D. Searle & Co., Monsanto Company filed Critical G.D. Searle & Co.
Priority to AU75186/94A priority Critical patent/AU7518694A/en
Publication of WO1995006061A1 publication Critical patent/WO1995006061A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06139—Dipeptides with the first amino acid being heterocyclic
    • C07K5/06165—Dipeptides with the first amino acid being heterocyclic and Pro-amino acid; Derivatives thereof
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/02—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link
    • C07K5/0207—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link containing the structure -NH-(X)4-C(=0), e.g. 'isosters', replacing two amino acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06008—Dipeptides with the first amino acid being neutral
    • C07K5/06017—Dipeptides with the first amino acid being neutral and aliphatic
    • C07K5/06026—Dipeptides with the first amino acid being neutral and aliphatic the side chain containing 0 or 1 carbon atom, i.e. Gly or Ala
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06008—Dipeptides with the first amino acid being neutral
    • C07K5/06017—Dipeptides with the first amino acid being neutral and aliphatic
    • C07K5/06034—Dipeptides with the first amino acid being neutral and aliphatic the side chain containing 2 to 4 carbon atoms
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • the present invention relates to selected retroviral protease inhibitors and combinations of retroviral protease inhibitors which are effective in preventing the replication of mammalian retroviruses, such as human immunodeficiency virus (HIV) . More particularly, the present invention relates to novel compounds, compositions, combinations of compounds and methods for inhibiting retroviral proteases.
  • This invention in particular, relates to urea-containing hydroxyethylamine protease inhibitor compounds, compositions, combinations of such compounds and method for inhibiting retroviral proteases, such as HIV protease, and for treatment or prophylaxis of retroviral infections, such as HIV infections.
  • the subject invention also relates to processes for making such compounds as well as to intermediates useful in such processes.
  • gag and gag-pol gene transcription products are translated as proteins. These proteins are subsequently processed by a virally encoded protease (or proteinase) to yield viral enzymes and structural proteins of the virus core.
  • the gag precursor proteins are processed into the core proteins and the pol precursor proteins are processed into the viral enzymes, e.g., reverse transcriptase and retroviral protease.
  • the retroviral protease It has been shown that correct processing of the precursor proteins by the retroviral protease is necessary for assembly of infectious virons. For example, it has been shown that frameshift mutations in the protease region of the pol gene of HIV prevents processing of the gag precursor protein. It has also been shown through site-directed mutagenesis of an aspartic acid residue in the HIV protease active site that processing of the gag precursor protein is prevented.
  • attempts have been made to inhibit viral replication by inhibiting the action of retroviral proteases.
  • Retroviral protease inhibition typically involves a transition-state mimetic whereby the retroviral protease is exposed to a mimetic compound which binds (typically in a reversible manner) to the enzyme in competition with the gag and gag-pol proteins to thereby inhibit specific processing of structural proteins and the release of retroviral protease itself. In this manner, retroviral replication proteases can be effectively inhibited.
  • mimetic compounds have been proposed, particularly for inhibition of proteases, such as for inhibition of HIV protease.
  • mimetics include hydroxyethylamine isosteres, reduced amide isosteres and non-peptide isosteres. See for example, EP 0 346 847; EP O 342,541; Roberts et al, "Rational Design of Peptide-Based Proteinase Inhibitors," Science. 248, 358 (1990) ; Erickson et al, "Design Activity, and 2.8A Crystal Structure of a C2 Symmetric Inhibitor
  • renin Several classes of mimetic compounds are known to be useful as inhibitors of the proteolytic enzyme renin. See, for example, U.S. No. 4,599,198; U.K. 2,184,730; G.B. 2,209,752; EP 0 264 795; G.B. 2,200,115 and U.S. SIR H725. Of these, G.B. 2,200,115, GB 2,209,752, EP O 264,795, U.S. SIR H725 and U.S. 4,599,198 disclose urea- containing hydroxyethylamine renin inhibitors.
  • the present invention is directed to virus inhibiting compounds and compositions. More particularly, the present invention is directed to retroviral protease inhibiting compounds, compositions and combinations of such compounds, to a method of inhibiting retroviral proteases, to processes for preparing the compounds and to intermediates useful in such processes.
  • the subject compounds are characterized a's urea-containing hydroxyethylamine inhibitor compounds.
  • R 1 is iso-propyl, sec-butyl, tert-butyl, -C(CH 3 )2(SCH 3 ), -C(CH 3 ) 2 (S ⁇ 0 ⁇ CH 3 ) or -C(CH 3 ) 2 (S ⁇ 0 ⁇ 2 CH 3 ) ; and R 2 is N-methyl-L-alaninyl, N-methyl-D-alaninyl, glycinyl, N-methylglycinyl, L-prolyl, D-prolyl or L- isoleucinyl, each of which is optionally substituted on the nitrogen atom with benzyloxycarbonyl or tert- butoxycarbonyl.
  • the present invention is also a method of treating a retroviral protease infection in a mammal, such as a human, monkey, cat and the like, suffering therefrom comprising administering a compound of the Formula (II) :
  • the compounds of this method preferably are of the Formula I and Formula II wherein R 1 is tert-butyl or sec-butyl; and R 2 is N-methylglycinyl.
  • the present invention relates to a novel compound, butaneamide, 2-[(N- monomethylaminoacetyl)amino] -N-[3- [ [ [ (1,1- dimethylethyl)amino]carbonyl] (3-methylbutyl)amino] -2- hydroxy-1-(phenyl ethyl)propyl] -3,3-dimethyl-, [1S- [1R*(R*) ) ,2S*] ] -, selected analogs and pharmaceutically acceptable salts thereof.
  • the novel compound may also be known as (2R,3S) -3 (N-methylaminoacetyl-L-tert- butylglycinyl)amido-l-isoamyl-1- (tert- butylcarbamoyl)amino-4-phenyl-2-butanol.
  • the structure of the novel compound is consistent with that of a metabolite obtained from its precursor or prodrug.
  • the prodrug is butaneamide, 2-[ (N,N-dimethylaminoacetyl) amino] -N-[3-[ [ [ (1, 1-dimethylethyl)amino]carbonyl] (3- methylbutyl)amino] -2-hydroxy-1-(phenylmethyl)propyl] - 3,3-dimethyl-, [IS-[1R* (R*) ) ,2S*] ] -, selected analogs and pharmaceutically acceptable salt thereof, which is disclosed in co-owned and co-pending U.S. Application Serial Number 08/152,934 (filed November 15, 1993) and WO Patent Application PCT/US91/08613, both incorporated herein by reference in their entirety, and is also shown in Example 1 hereinafter.
  • R 1 is as defined above, is obtained by administering a retroviral protease inhibitor of Formula II as defined above to a mammal, collecting blood samples from the mammal, and separating the metabolite from the -sample (see Example 2 infra) .
  • Formula II is a precursor or prodrug.
  • Suitable laboratory mammals from which the metabolites of the present invention can be obtained include dog, rat and the like.
  • the compounds of the Formula I of the present invention can be prepared utilizing the following general procedure.
  • Z represents an amino protecting group
  • Suitable amino protecting groups are well known in the art and include haloacetyl or more specifically chloroacetyl, carbobenzoxy, butyryl, t-butoxycarbonyl, acetyl, benzoyl and the like.
  • a preferred amino protecting group is carbobenzoxy or more preferably chloroacetyl.
  • a preferred N-protected chloroketone is N-benzyloxycarbonyl-L-phenylalanine chloromethyl ketone.
  • a preferred reducing agent is sodium borohydride.
  • the reduction reaction is conducted at a temperature of from -10°C to about 25°C, preferably at about 0°C, in a suitable solvent system such as, for example, tetrahydrofuran, and the like.
  • a suitable solvent system such as, for example, tetrahydrofuran, and the like.
  • the N-protected chloroketones are commercially available from Bachem, Inc., Torrance, California. Alternatively, the chloroketones can be prepared by the procedure set forth in S. J. Fittkau, J. Prakt. Chem. , 315, 1037 (1973), and subsequently N-protected utilizing procedures which are well known in the art.
  • the resulting alcohol is then reacted, preferably at room temperature, with a suitable base in a suitable solvent system to produce an N-protected amino epoxide of the Formula (2)
  • Suitable solvent systems for preparing the amino epoxide include ethanol, methanol, isopropanol, tetrahydrofuran, dioxane, and the like including mixtures thereof.
  • Suitable bases for producing the epoxide from the reduced chloroketone include potassium hydroxide, sodium hydroxide, potassium t-butoxide, DBU and the like. A preferred base is potassium hydroxide. / ' Although only one of the diastereomers is illustrated, products may be diastereomeric mixtures. The diastereomers can be separated by chromatography or, alternatively, once reacted in subsequent steps the diastereomeric products can be separated.
  • the amino epoxide is then reacted, in a suitable solvent system, with an equal amount, or preferably an excess of, isoamylamine.
  • the reaction can be conducted over a wide range of temperatures, e.g., from about 10°C to about 100°C, but is preferably, but not necessarily, conducted at a temperature at which the solvent begins to reflux.
  • Suitable solvent systems include those wherein the solvent is an alcohol, such as methanol, ethanol, isopropanol, and the like, ethers such as tetrahydrofuran, dioxane and the like, and toluene, N,N- dimethylformamide, dimethyl sulfoxide, and mixtures thereof.
  • a preferred solvent is isopropanol.
  • the resulting product is the following protected amino alcohol
  • Suitable solvent systems include tetrahydrofuran, methylene chloride, and the like and mixtures thereof.
  • the resulting product is a urea derivative of the amino alcohol and can be represented by the Formula (3)
  • the tert-butylisocyanate can be prepared by the reaction of a tert-butylamine with phosgene, triphosgene, carbodiimidazole, or carbonate ( (RO) 2 C0) under conditions well-known in the art.
  • the tert-butylisocyanate is commercially available from Aldrich Chemical Company or may be prepared by using methods well known to those skilled in the art, e.g., an appropriate carboxylic acid and the Curtius rearrangement.
  • the amino protecting group is removed under conditions which will not affect the remaining portion of the molecule.
  • These methods include acid hydrolysis, hydrogenolysis and the like.
  • a preferred method involves removal of the protecting group, e.g., removal of a carbobenzoxy group, by hydrogenolysis utilizing palladium on carbon in a suitable solvent system such as an alcohol, acetic acid, and the like or mixtures thereof.
  • the protecting group is a t- butoxycarbonyl group, it can be removed utilizing an inorganic or organic acid, e.g., HCl or trifluoroacetic acid, in a suitable solvent system, e.g., dioxane or methylene chloride.
  • the resulting product is the amine salt derivative.
  • the amine is then coupled to an amino acid, for example, Z-t-butylglycine or corresponding derivative thereof represented by the formula (ZNHCH(R 1 )COOH) , an etamino acid, wherein Z and R ⁇ are as defined above, using standard peptide coupling methods.
  • an amino acid for example, Z-t-butylglycine or corresponding derivative thereof represented by the formula (ZNHCH(R 1 )COOH)
  • an etamino acid wherein Z and R ⁇ are as defined above.
  • Such ecamino acids are well known, are commercially available or can be prepared by known methods. This produces the intermediate compounds of the present invention having t'he Formula (4)
  • Z and R 1 are as defined above.
  • Preferred protecting groups in this instance are a benzyloxycarbonyl group or a t-butoxycarbonyl group. Removal of the Z protecting group from a compound of Formula 4 using conditions well known to those skilled in the art, for example, catalytic hydrogenolysis using palladium or palladium on carbon as catalysts, provides the intermediate amine. Then the intermediate amine obtained from the Formula 4 is reacted with chloroacetic anhydride to obtain the intermediate with the Formula (5)
  • the Z protecting group may be a chloroacetyl group which would eliminate the deprotection and' acylation steps above.
  • the compound of Formula 6 is obtained by reacting the compound of Formula 5 with monomethylamine.
  • Appropriate solvent systems for such a reaction include tetrahydrofuran, acetonitrile, methylene chloride, N,N- dimethylformamide, or alcohols, such as methanol, ethanol or isopropanol and the like, including mixtures thereof.
  • Formula 4 may be coupled with a Z protected N-methylglycine using standard peptide coupling methods.
  • the compound of Formula 6 is then obtained by deprotecting the amine using the methods discussed above.
  • the compounds of Formula I where R 1 is a sulfoxide or sulfone derivative of L-penicillamine can also be prepared by oxidation of the L-penicillamine derivative of Formula I or its nitrogen protected (Z) derivative.
  • Reagents to accomplish such oxidations are well known to those skilled in the art and include, for example, one or two equivalents of hydrogen peroxide, peracetic acid, meta-chloroperbenzoic acid, percamphoric acid, a metal salt of periodate or periodic acid or the like.
  • Temperatures for the reaction can range from about -22°C to about 60°C preferably about 0°C in the case of the sulfoxide.
  • Solvents for the transformation include water, alcohols, dipolar aprotic solvents such as DMF, acetonitrile, nitromethane or non-protic solvents such as THF, methylene chloride, ethyl acetate and the like or mixtures of the above.
  • Preferred conditions for preparation of the sulfoxides or their salts are sodium metaperiodate or periodic acid in water or mixtures of water with alcohols such as methanol, ethanol, denatured ethanol, isopropanol and the like carried at about 0°C.
  • Scheme I is illustrative of the preparation of the compounds of the present invention.
  • a protected amino epoxide of Formula (7) can also be used in the preparation of the compounds of the present invention:
  • P 1 and P 2 are each independently amine protecting groups, including but not limited to arylalkyl, substituted arylalkyl, cycloalkenylalkyl, substituted cycloalkenylalkyl, allyl, substituted allyl, acyl, alkoxycarbonyl, aralkoxycarbonyl or silyl.
  • arylalkyl examples include, but are not limited to benzyl, ortho-methylbenzyl, trityl and benzhydryl, which can be optionally substituted with halogen, alkyl of C ⁇ -
  • aryl groups include phenyl, naphthalenyl, indanyl, anthracenyl, durenyl, 9- (9-phenylfluorenyl) and phenanthrenyl, cycloalkenylalkyl or substituted cycloalkylenylalkyl radicals containing cycloalkyls of C 8 -C 10 .
  • Suitable acyl groups include carbobenzoxy, t-butoxycarbonyl, iso-butoxycarbonyl, benzoyl, substituted benzoyl, butyryl, acetyl, tri- fluoroacetyl, tri-chloroacetyl, phthaloyl and the like.
  • the 'term silyl refers to a silicon atom optionally substituted by one or more alkyl, aryl and aralkyl groups.
  • Suitable silyl protecting groups include, but are not limited to, trimethylsilyl, triethylsilyl, tri- isopropylsilyl, tert-butyldimethylsilyl, dimethylphenylsilyl, 1,2-bis(dimethylsilyl)benzene, 1,2- bis(dimethylsilyl)ethane and diphenylmethylsilyl.
  • Silylation of the amine functions to provide mono- or bis-disilylamine can provide derivatives of the aminoalcohol, amino acid, amino acid esters and amino acid amide. In the case of amino acids, amino acid esters and amino acid amides, reduction of the carbonyl function provides the required mono- or bis-silyl aminoalcohol.
  • Silylation of the aminoalcohol can lead to the N,N,0-tri-silyl derivative.
  • Removal of the silyl function from the silyl ether function is readily accomplished by treatment with, for example, a metal hydroxide or ammonium flouride reagent, either as a discrete reaction step or in situ during the preparation of the amino aldehyde reagent.
  • Suitable silylating agents are, for example, trimethylsilyl chloride, tert- buty-dimethylsilyl chloride, phenyldimethylsilyl chlorie, diphenylmethylsilyl chloride or their combination products with imidazole or DMF.
  • P 1 and P 2 can form a heterocyclic ring with the nitrogen to which they are attached, for example, 1,2- bis (methylene)benzene, phthalimidyl, succinimidyl, maleimidyl and the like and where these heterocyclic groups can further include adjoining aryl and cycloalkyl rings.
  • the heterocyclic groups can be mono-, di- or tri-substituted, e.g., nitrophthalimidyl.
  • the economical and safe large scale method of preparation of protease inhibitors of the present invention can alternatively utilize amino acids or amino alcohols to form N,N-protected alpha aminoalcohol of the Formula (8)
  • the amine protecting groups P 1 and P2 are introduced by alkylation of the amine group such as by the addition of suitable alkylating agents in an appropriate solvent in the presence of base.
  • bases used in alkylation include sodium hydroxide, sodium bicarbonate, potassium hydroxide, lithium hydroxide, potassium carbonate, sodium carbonate, cesium hydroxide, magnesium hydroxide, calcium hydroxide or calcium oxide, or tertiary amine bases such as triethyl amine, diisopropylethylamine, N-methylpiperidine, pyridine, dimethylaminopyridine and azabicyclononane. Reactions can be homogenous or heterogenous.
  • Suitable solvents are water and protic solvents or solvents miscible with water, such as methanol, ethanol,
  • Dipolar aprotic solvents may also be ised with or without added protic solvents including water.
  • Examples of dipolar aprotic solvents include acetonitrile, dimethylformamide, dimethyl acetamide, acetamide, tetramethyl urea and its cyclic analog, dimethylsulfoxide, N-methylpyrrolidone, sulfolane, nitromethane and the like.
  • Reaction temperature can range between about -20° to 100°C with the preferred temperature of about 25-85°C.
  • the reaction may be carried out under an inert atmosphere such as nitrogen or argon, or normal or dry air, under atmospheric pressure or in a sealed reaction vessel under positive pressure.
  • the most preferred alkylating agents are benzyl bromide or benzyl chloride or monosubstituted aralkyl halides or polysubstituted aralkyl halides.
  • Sulfate or sulfonate esters are also suitable reagents to provide the corresponding benzyl analogs and they can be preformed from the corresponding benzyl alcohol or formed in situ by methods well known to those skilled in the art.
  • Trityl, benzhydryl, substituted trityl and substituted benzhydryl groups are also effective amine protecting groups as are allyl and substituted allyl groups.
  • Their halide derivatives can also be prepared from the corresponding alcohols by methods well known to those skilled in the art such as treatment with thionyl chloride or bromide or with phosphorus tri- or pentachloride, bromide or iodide or the corresponding phosphoryl trihalide.
  • Examples of groups that can be substituted on the aryl ring include alkyl, alkoxy, hydroxy, nitro, halo and alkylene, amino, mono- and dialkyl amino and acyl amino, acyl and water solubilizing groups such as phosphonium salts and ammonium salts.
  • the aryl ring can be derived from, for example, benzene, napthelene, indane, anthracene, 9-(9- phenyl fluorenyl, durene, phenanthrene and the like.
  • 1,2-bis (substituted alkylene) aryl halides or sulfonate esters can be used to form a nitrogen containing aryl or non-aromatic heterocyclic derivative [with P 1 and P ] or bis-heterocycles.
  • Cycloalkylenealkyl or substituted cyloalkylene radicals containing 6-10 carbon atoms and alkylene radicals constitute additional acceptable class of substituents on nitrogen prepared as outlined above including, for example, cyclohexylenemethylene.
  • Compounds of Formula 8 can also be prepared by reductive alkylation by, for example, compounds and intermediates formed from the addition of an aldehyde with the amine and a reducing agent, reduction of a Schiff Base, carbinolamine or enamine or reduction of an acylated amine derivative.
  • Reducing agents include metals, such as platinum, palladium, palladium hydroxide, palladium on carbon, platinum oxide, rhodium and the like, in the presence of hydrogen gas or hydrogen transfer molecules such as cyclohexene, cyclohexadiene and the like, or hydride agents such as lithium aluminumhydride, sodium borohydride, lithium borohydride, sodium cyanoborohydride, diisobutylaluminum hydride, lithium tri-tert-butoxyaluminum hydride and the like.
  • metals such as platinum, palladium, palladium hydroxide, palladium on carbon, platinum oxide, rhodium and the like
  • hydrogen gas or hydrogen transfer molecules such as cyclohexene, cyclohexadiene and the like
  • hydride agents such as lithium aluminumhydride, sodium borohydride, lithium borohydride, sodium cyanoborohydride, diisobutylaluminum hydride, lithium tri-tert-
  • Additives such as sodium or potassium bromide, sodium or potassium iodide can catalyze or accelerate the rate of amine alkylation, especially when benzyl chloride was used as the nitrogen alkylating agent.
  • Phase transfer catalysis wherein the amine to be protected and the nitrogen alkylating agent are reacted with base in a solvent mixture in the presence of a phase transfer reagent, catalyst or promoter.
  • the mixture can consist of, for example, toluene, benzene, ethylene dichloride, cyclohexane, methylene chloride or the like with water or a aqueous solution of an organic water miscible solvent such as THF.
  • phase transfer catalysts or reagents examples include tetrabutylammonium chloride or iodide or bromide, tetrabutylammonium hydroxide, tri-butyloctylammonium chloride, dodecyltrihexylammonium hydroxide, methyltrihexylammonium chloride and the like.
  • a preferred method of forming substituted amines involves the aqueous addition of about 3 moles of organic halide to the amino acid or about 2 moles to the aminoalcohol. In a more preferred method of forming a protected amino alcohol, about 2 moles of benzylhalide in a basic aqueous solution is utilized.
  • the alkylation occurs at 50"C to 80 * C with potassium carbonate in water, ethanol/water or denatured ethanol/water.
  • about 3 moles of benzylhalide is added to a solution containing the amino acid.
  • the protected amino acid ester is reduced to the protected amino alcohol in an organic solvent.
  • Preferred reducing agents include lithium aluminiumhydride, lithium borohydride, sodium borohydride, borane, lithium tri-ter-butoxyaluminum hydride, borane*THF complex and the like.
  • the reducing agent is diisobutylaluminum hydride (DiBAL-H) in toluene. These reduction conditions provide an alternative to a lithium aluminum hydride reduction.
  • the protected alpha amino alcohol is oxidized to form a chiral amino aldehyde of the Formula (9)
  • Acceptable oxidizing reagents include, for example, sulfur trioxide-pyridine complex and DMSO, oxalyl chloride and
  • a preferred oxidation method is sulfur trioxide pyridine complex, triethylamine and DMSO at room temperature. This system provides excellent yields of the desired chiral protected amino aldehyde usable without the need for purification by chromatography and large scale operations are made less hazardous. Reaction at room temperature also eliminates the need for the use of low temperature conditions.
  • the reaction may be carried out under and inert atmosphere such as nitrogen or argon, or normal or dry air, under atmospheric pressure or in a sealed reaction vessel under positive pressure.
  • Preferred is a nitrogen atmosphere.
  • Alternative amine bases include, for example, tri-butyl amine, tri-isopropyl amine, N- methylpiperidine, N-methyl morpholine, azabicyclononane, diisopropylethyla ine, 2,2,6,6-tetramethylpiperidine, N,N-dimethylaminopyridine, or mixtures of these bases.
  • Triethylamine is a preferred base.
  • DMSO dimethylformamide
  • dimethylacetamide dimethylacetamide
  • acetamide tetramethyl urea and its cyclic analog
  • N-methylpyrrolidone sulfolane and the like.
  • the phenylalaninol derivatives discussed above can be used to provide the corresponding N-monosubstituted or N,N-disubstituted aldehyde.
  • hydride reduction of an amide or ester derivative of the corresponding alkyl, benzyl or cycloalkenyl nitrogen protected phenylalanine, substituted phenylalanine or cycloalkyl analog of phenylalanine derivative can be carried out to provide a compound of Formula 8. Hydride transfer is an additional method of aldehyde synthesis under conditions where aldehyde condensations are avoided, cf, Oppenauer Oxidation.
  • the aldehydes of this process can also be prepared by methods of reducing protected phenylalanine and phenylalanine analogs or their amide or ester derivatives by, e.g., sodium amalgam with HCl in ethanol or lithium or sodium or potassium or calcium in ammonia.
  • the reaction temperature may be from about -35 * C to about 45"C, and preferably from about 5 * C to about 25 * C or when the concentration of ammonia approaches 100% the temperature is preferably about -33°C.
  • Two additional methods of obtaining the nitrogen protected aldehyde include oxidation of the corresponding alcohol with bleach in the presence of a catalytic amount of 2,2,6,6- tetramethyl-1-pyridyloxy free radical.
  • oxidation of the alcohol to the aldehyde is accomplished by a catalytic amount of tetrapropylammonium perruthenate in the presence of N- methylmorpholine-N-oxide.
  • an acid chloride derivative of a protected phenylalanine or phenylalanine derivative as disclosed above can be reduced with hydrogen and a catalyst such as Pd on barium carbonate or barium sulphate, with or without an additional catalyst moderating agent such as sulfur or a thiol (Rosenmund Reduction) .
  • a catalyst such as Pd on barium carbonate or barium sulphate
  • an additional catalyst moderating agent such as sulfur or a thiol (Rosenmund Reduction) .
  • An important aspect for this preparation of an appropriate amino-epoxide useful as an intermediate to prepare a compound of the present invention is a rea ' ction involving the addition of chloromethylithium or bromomethyllithium to the ceamino aldehyde, e.g. addition of chloromethyllithium or bromomethylithium to racemic or chiral amino aldehydes to form aminoepoxides of the Formula 10
  • chloromethylithium or bromomethylithium to a chiral amino aldehyde is highly diastereoselective.
  • the chloromethyllithium or bromomethylithium is generated in-situ from the reaction of the dihalomethane and n-butyllithium.
  • Acceptable methyleneating halomethanes include chloroiodomethane, bromochloromethane, dibromomethane, diiodomethane, bromofluoromethane and the like.
  • the sulfonate ester of the addition product of, for example, hydrogen bromide to formaldehyde is also a methyleneating agent.
  • Tetrahydrofuran is the preferred solvent, however alternative solvents such as toluene, dimethoxyethane, ethylene dichloride, methylene chloride can be used as pure solvents or as a mixture.
  • Dipolar aprotic solvents such as acetonitrile, DMF, N-methylpyrrolidone are useful as solvents or as part of a solvent mixture.
  • the reaction can be carried out under an inert atmosphere such as nitrogen or argon.
  • n-butyl lithium can be substituted other organometallic reagents such as methyllithium, tert-butyl lithium, sec-butyl lithium, phenyllithium, phenyl sodium and the like.
  • the reaction can be carried out at temperatures of between about -80 * C to 0 * C but preferably between about -80 * C to -10"C.
  • the most preferred reaction temperatures are between -40 * C to -15"C.
  • Reagents can be added singly but multiple additions are preferred in certain conditions.
  • the preferred pressure of the reaction is atmospheric however a positive pressure is valuable under certain conditions such as a high humidity environment.
  • Alternative methods of conversion to the epoxides of this invention include substitution of other charged methylenation precursor species followed by their treatment with base to form the analogous anion.
  • these species include trimethylsulfoxonium tosylate or triflate, tetramethylammonium halide, methyldiphenylsulfoxonium halide wherein halide is chloride, bromide or iodide.
  • the conversion of the aldehydes of this invention into their epoxide derivative can also be carried out in multiple steps.
  • the protected aminosulfide alcohol can be alkylated with, for example, the alkylating agents above, to provide a sulfonium salts that are subsequently converted into the subject epoxides with tert-amine or mineral bases.
  • the desired epoxides form, using most preferred conditions, diastereoselectively in ratio amounts of at least about an 85:15 ratio (S,S:S,R).
  • the product can be purified by chromatography to give the diastereomerically and enantiomerically pure product but it is more conveniently used directly without purification to prepare the HIV protease inhibitors of the present invention.
  • the diastereomers can be separated by chromatography or, alternatively, once reacted in subsequent steps the diastereomeric products can be separated.
  • the amino epoxide is then reacted, in a suitable solvent system, with at least an equal amount, preferably an excess, of isoamylamine.
  • the reaction can be conducted over a wide range of temperatures, e.g., from about 10 * C to about 100 * C, but is preferably, but not necessarily, conducted at a temperature at which the solvent begins to reflux.
  • Suitable solvent systems include those wherein the solvent is an alcohol, such as methanol, ethanol, isopropanol, and the like, ethers such as tetrahydrofuran, dioxane and the like, and toluene, N,N- dimethylformamide, dimethyl sulfoxide, and mixtures thereof.
  • a preferred solvent is isopropanol.
  • the resulting product is a protected amino alcohol.
  • This protected amino alcohol can then be reacted, with tertiary-butylisocyanate and the protecting groups, P and P , removed using methods well known to those skilled in the art.
  • P 1 and P 2 are both benzyl groups, these can be removed by hydrogenolysis using a palladium catalyst.
  • the resulting amine can then be converted into the desired product through the methods outlined in Scheme 1 above or in an analogous manner for the anaolgous and pharmaceutically acceptable salts of the invention.
  • the chemical reactions described above are generally disclosed in terms of their broadest application to the preparation of the compounds of this invention.
  • Step A N-Cbz-3- (S)-amino-l-chloro-4-phenvl-2 (S)- butariQ-1
  • Step B N-Cbz-3- (S) -amino-1.2-(S1 -enoxv- -nhenvlbutane
  • Step C N-r T I S ) -nhenv1 ⁇ nethylcarbamoyl)amino-2 (R) - hvdroxv-4-phenvlbut-.vll -N- r (3-methylbutvl) 1amine
  • N-Cbz-3-(S) -amino-1,2-(S) -epoxy-4-phenylbutane (10 gm, 33.6 mmol) and isoamylamine (19.5 mL, 168 mmol) were combined in isopropanol (100 mL) and stirred for 16 hours at room temperature. The insoluble material was collected by filtration, washed with a small amount of isopropanol and then n-hexane.
  • Step D (2R. 3S) -3-(phenvlmethoxvcarbonvl) mino-2- hvdroxy-4-phenyl-l-TN-(3-methylbutyl) -N-f (1.1- dimethylethyl)amino1carbonyll aminobutane
  • Step E (2R. 3S) -3-amino-2-hvdroxy-4-ohenyl-l-fN-(3- methvlbutvl)-N-r(l.l- dime hvlethvl)amino1carbonvllaminobutane
  • Step F (2R. 3S) -3- (N-benzvloxvcarbonvl-L-tert- butvl ⁇ lvciinvl) amino-2-hvdroxv-4-phenvl-l-TN-(3- methylbutyl) -N-r (1.1- dimethvlethvl)amino1carbonvllaminobutane
  • Step G (2R. 3S) -3- (L-tert-butvl ⁇ lvcinvl) mino-2- hvdroxv-4-phenvl-l-TN-( -methvlbutvl ) -N-r (1.1- dimethylethyl)amino1carbonyll aminobutane
  • Step I (2R, 3S) -3 (N-methylaminoacetyl-L-tert- butvl ⁇ lvcinvl) amJno-2-hvdroxv-4-r)henvl-l- TN-(3- methylbutyl) -N- U l .1-dimethylethyl)amino! carbonvllaminobutane or butaneamide.
  • 2-r N-monomethvlaminoacetvl) mino! -N-T3- r r r (l.l-dimethvlethvl)amino!carbonvll (3- methylbutyl)amino! -2-hvdroxy-l-(phenylmethyl)prooyll - 3.3-dimethyl-.
  • Step J (2R. 3S) - (N-methvlaminoacetvl-L-tert- butvl ⁇ lvcinvl)amino-2-hvdroxv-4-phenvl-l-rN-(3- methvlbutvl) -N-r (1,1-dimethylethvl)amino!carbonvll aminobutane hvdrochloride or butaneamide.
  • Step A (2R.3S) -3 (N-tert-butvloxvcarbonvl-L- isoleucinvl)amido-1-rN-isoamvl-N- (tert- butvlcarbamovl) 1amino-4-phenvl-2-butanol
  • Step B (2R.3S) -3- (L-isoleucinyl)amido-1-rN-isoamyl-N- (tert-butvlcarbamovl) 1amino-4-phenvl-2-butanol hyfa-pchlqrifle ?elt
  • Step C (2R.3S)-3-(N-chloroacetvl-T,-isoleucinvl)amido- 1-rN-isoamvl-N-(tert-butvlcarbamovl) 1amino-4-phenvl-2- butanol
  • Step D (2R.3S) -3- (N-methvlaminoacetvl -L-isoleucinvl) phenvl-2-butanol or butaneamide.
  • Step A (2R. 3S)-3-(N-tert-butvloxvcarbonvl-S-methvl-L- penicillaminvl)amido-l-rN-isoamvl-N-(tert- butylcarbamoyl) 1amino-4-phenyl-2-butanol
  • Step B (2R. 3S)-3-(S-me hvl-L-penicillaminvl)amido-l- rN-isoamvl-N-(tert-butvlcarbamovl) 1 mino-4-phenvl-2- butanol hydrochloride
  • Step C f2R. 3S) -3-(N-chloroacetvl-S-methvl-L- penicillaminvl) amido-1-rN-isoamvl-N-(tert- butylcarba oyl) 1amino-4-phenyl-2-butanol
  • Step D (2R. 3S) -3-(N-methylaminoacetyl-S-methyl-L- penicillaminvl)amido-l-fN-isoamvl-N-(tert- butvlcarbamovl) 1 mino-4-phenvl-2-butanol
  • Examples 7-9 exemplify the more preferred method of preparing the N,N, alpha-S- Tris (phenylmethyl) -2S-oxiranemethanamine, useful as an intermediate in the described general procedure.
  • the solid material was suspended in a mixture of toluene (400 mL) and water (100 ml) .
  • the mixture was cooled to 5"C and treated with 2.5 N NaOH (186 mL) and then stirred at room temperature until solid dissolved.
  • the toluene layer was separated from the aqueous phase and washed with water and brine, dried over magnesium sulfate, filtered and concentrated to a volume of 75 mL (89 g) .
  • Ethyl acetate (25 mL) and hexane (25 mL) were added to the residue upon which the desired alcohol product began to crystallize. After 30 min, an additional 50 mL hexane were added to promote further crystallization.
  • L-phenylalaninol (176.6 g, 1.168 mol) was added to a stirred solution of potassium carbonate (484.6 g, 3.506 mol) in 710 mL of water. The mixture was heated to 65 * C under a nitrogen atmosphere. A solution of benzyl bromide (400 g, 2.339 mol) in 3A ethanol (305 mL) was added at a rate that maintained the temperature between 60-68 * C. The biphasic solution was stirred at 65 * C for 55 ,min and then allowed to cool to lO'C with vigorous stirring. The oily product solidified into small granules. The product was diluted with 2.0 L of tap water and stirred for 5 minutes to dissolve the inorganic by products.
  • the reaction mixture was cooled with ice water and quenched with 1.6 L of cold water (10-15 * 0 over 45 minutes.
  • the resultant solution was' extracted with ethyl acetate (2.0 L) , washed with 5% citric acid (2.0 L) , and brine (2.2 L) , dried over MgS04 (280 g) and filtered.
  • the solvent was removed on a rotary evaporator at 35-40 * C and then dried under vacuum to give 198.8 g of alpha-S-[Bis-(phenylmethyl)amino] - benzenepropanaldehyde as a pale yellow oil (99.9%).
  • the crude product obtained was pure enough to be used directly in the next step without purification.
  • Triethylamine (41.2 ml, 0.295 mol) was then added over 10 min. (temp. -78 "to -68 * C) upon which the ammonium salt procipitated.
  • the cold mixture was stirred for 30 min. and then water (225 ml) was added.
  • the dichloromethane layer was separated from the aqueous phase and washed with water, brine, dried over magnesium sulfate, filtered and concentrated. The residue was diluted with ethyl acetate and hexane and then filtered to further remove the ammonium salt. The filtrate was concentrated to give the desired aldehyde product.
  • the aldehyde was carried on to the next step without purification.
  • the mixture was stirred for 10 minutes and theorganic and aqueous layers were separated.
  • the aqueous phase was extracted with ethyl acetate (2x 500 mL) .
  • the ethyl acetate layer was combined with the tetrahydrofuran layer.
  • the combined solution was dried over magnesium sulfate (220 g) , filtered and concentrated on a rotary evaporator at 65"C.
  • the brown oil residue was dried at 70"C in vacuo (0.8 bar) for 1 h to give 222.8 g of crude material. (The crude product weight was >100%. Due to the relative instability of the product on silica gel, the crude product is usually used directly in the next step without purification) .
  • the diastereomeric ratio of the crude mixture was determined by proton NMR: (2 ⁇ )/(2R) : 86:14.
  • An analytical sample of each of the diastereomers was obtained by purification on silica-gel chromatography (3% ethyl acetate/hexane) and characterized as follows:
  • the mixture was stirred for 10 minutes and theorganic and aqueous layers were separated.
  • the aqueous phase was extracted with ethyl acetate (2x 500 mL) .
  • the ethyl acetate layer was combined with the tetrahydrofuran layer.
  • the combined solution was dried over magnesium sulfate (220 g) , filtered and concentrated on a rotary evaporator at 65 * C.
  • the brown oil residue was dried at 70 * C in vacuo (0.8 bar) for 1 h to give 222.8 g of crude material.
  • the desired product may be obtained in a manner similar to methods 1 and 2 above.
  • the catalyst is removed by filtration through a sintered glass funnel and concentrated to an off white foam. Final traces of solvent are removed in-vacuo to provide 98.3g (101%) of (2R,3S)-3-amino-l-isoamyl-l-(tert-butylcarbamoyl)amino- 4-phenyl-2-b ⁇ tanol, the compound of Example 2, Step E above.
  • Example 19 The product of Example 19 above (205 mg, 0.3 mmol) was dissolved in MeOH (15 mL) and to this was added 10% Pd on carbon. The mixture was hydrogenated at 5 psi of hydrogen at ambient temperature for 4 hours. The reaction mixture was filtered through diatomaceous earth and concentrated to a gummy solid, 160 mg (95%) . The product was identified by its NMR spectrum..
  • Butaneamide, 2-[ (N,N-dimethylaminoacetyl)amino] -N- [3-[ [ t (1/1-dimethylethyl)amino]carbonyl] (3- methylbutyl)amino] -2-hydroxy-1-(phenylmethyl)propyl] - 3,3-dimethyl-, [IS-[1R*(R*) , 2S*]]-, as prepared in Example 1 herein, is administered to male rats (n 5) orally in water containing 0.5% methylcellulose and 0.1% polysorbate 80 in a total volume of 5 to 7 mL per kilogram. Blood samples are collected at 0.25, 0.50, 1,
  • the samples for each time-point are pooled and extracted onto a 100 mg C 18 solid phase extraction column (SPEC) after prior treatment with methanol to denature plasma proteins.
  • the extract is purified by washing the SPEC with water and collected after the addition of 1% formic acid in methanol to the SPEC. Concentration of the extract is achieved by evaporation and resolubilization in a small volume of water:acetonitrile (3:1, v/v).
  • the resolubilized extract is filtered through a 0.2 ⁇ filter and injected onto a liquid chromatograph coupled to a mass spectrometer, using a VG Trio 2 Mass Spectrometer, by means of a thermospray interface.
  • the chromatographic separation is produced by use of a 10 cm (2 x 5 cm) YMC Basic column with a mobile phase of water:acetonitrile.-pyridine:formic acid (545:440:10:5, by volume) . Data are obtained for which the corresponding spectra and chromatogram from a standard of the synthesized metabolite can be favorably compared.
  • the compounds of the present invention are advantageously effective HIV protease inhibitors. Utilizing an enzyme assay as described below, the compounds set forth in the examples herein disclosed inhibited the HIV enzyme.
  • the preferred compounds of the present invention and their calculated IC50 (the concentration at which the inhibitor compound reduces enzyme activity by 50%) values are shown in Table 2.
  • the enzyme method is as follows.
  • the substrate is 2-aminobenzoyl-Ile-Nle-Phe(p-N02) -Gln-ArgNH2.
  • the positive control is MVT-101 (Miller, M. et al, Science, 246, 1149 (1989)).
  • the assay buffer is 20 mM sodium phosphate, pH 6.4, 20% glycerol, 1 mM EDTA, 1 mM DTT and 0.1% CHAPS.
  • the substrate is dissolved in DMSO, then diluted 10 fold in assay buffer. Final substrate concentration in the assay is about 80 ⁇ M.
  • HIV protease is diluted in the assay buffer to a final enzyme concentration of about 12.3 nanomolar, based on a molecular weight of 10,780.
  • the final concentration of DMSO is about 14% and the final concentration of glycerol is about 18%.
  • the tes.t compound is dissolved in DMSO and diluted in DMSO to ten times (lOx) the test concentration.
  • Ten microliters (10 ⁇ D of the enzyme preparation is added, the materials mixed and then the mixture is incubated at ambient temperature for 15 minutes.
  • the enzyme reaction is initiated by the addition of 40 ⁇ L of substrate.
  • the increase in fluorescence is monitored at 4 time points (0, 8, 16 and 24 minutes) at ambient temperature. Each assay is carried out in duplicate wells.
  • the HIV inhibition assay method of acutely infected cells is an automated tetrazolium based calorimetric assay essentially that reported by Pauwles et al, J. Virol. Methods 20, 309-321 (1988). Assays were performed in 96-well tissue culture plates. CEM cells, a CD4+ cell line, were grown in RPMI-1640 medium (Gibco) supplemented with a 10% fetal calf serum and were then treated with polybrene (2 ⁇ g/ml) . An 80 ⁇ l volume of medium containing 1 x 104 cells was dispensed into each well of the tissue culture plate.
  • tissue culture medium or medium without test compound as a control
  • TCID50 the dose of virus that infects 50% of cells in tissue culture
  • 20 ⁇ L volume of the virus sample was added to wells containing test compound and to wells containing only medium (infected control cells) .
  • TCID50 the dose of virus that infects 50% of cells in tissue culture
  • Several wells received culture medium without virus (uninfected control cells) .
  • the intrinsic toxicity of the test compound was determined by adding medium without virus to several wells containing test compound.
  • the tissue culture plates contained the following experiments:
  • test compounds were 1, 10, 100 and 500 ⁇ g/ml. Either azidothymidine (AZT) or dideoxyinosine (ddl) was included as a positive drug control. Test compounds were dissolved in DMSO and diluted into tissue culture medium so that the final DMSO concentration did not exceed 1.5% in any case. DMSO was added to all control wells at an appropriate concentration.
  • test compounds could be added on days 0, 2 and 5 if desired.
  • day 7 post-infection, the cells in each well were resuspended and a lOO ⁇ l sample of each cell suspension was removed for assay.
  • a 20 ⁇ L volume of a 5 mg/ l solution of 3-(4,5-dimethylthiazol-2-yl) -2,5- diphenyltetrazolium bromide (MTT) was added to each lOO ⁇ L cell suspension, and the cells were incubated for 4 hours at 27 * C in a 5% ' CO 2 environment.
  • MTT is metabolically reduced by living cells resulting in the production in the cell of a colored formazan product.
  • To each sample was added lOO ⁇ l of 10% sodium dodecylsulfate in 0.01 N HCl to lyse the cells, and samples were incubated overnight. The absorbance at 590 nm was determined for each sample using a Molecular Devices microplate reader. Absorbance values for each set of wells is compared to assess viral control infection, uninfected control cell response as well as test compound by cytotoxicity and antiviral efficacy.
  • the compounds of the present invention are advantageously effective antiviral compounds and, in particular, are effective inhibitors of retroviruses, particularly, lentiviruses as shown above.
  • the subject compounds are effective inhibitors of HIV. It is contemplated that the subject compounds will also inhibit other strains of HIV, such as HIV-2 and other viruses such as, for example, VIS-A virus and Simian Immunodeficiency virus (SIV) , HTLV-1 and HTLV-2.
  • the subject compounds are effective in the treatment and/or prophylaxis of retroviral infections.
  • optical isomers can be obtained by resolution of the racemic mixtures according to conventional processes, for example by formation of diastereoisomeric salts by treatment with an optically active acid or base.
  • appropriate acids are tartaric, diacetyltartaric, dibenzoyltartaric, ditoluoyltartaric and camphorsulfonic acid and then separation of the mixture of diastereoisomers by crystallization followed by liberation of the optically active bases from these salts.
  • a different process for separation of optical isomers involves the use of a chiral chromatography column optimally chosen to maximize the separation of the enantio ers.
  • Still another available method involves synthesis of covalent diastereoisomeric molecules by reacting compounds of Formula I with an optically pure acid in an activated form or an optically pure isocyanate.
  • the synthesized diastereoisomers can be separated by conventional means such as chromatography, distillation, crystallization or sublimation, and then hydrolyzed to deliver the enantiomericaly pure compound.
  • the optically active compounds of Formula I can likewise be obtained by utilizing optically active starting materials. These isomers may be in the form of a free acid, a free base, an ester or a salt.
  • the present invention is also meant to include the solvate or hydrates of the compounds of the formula I, when possible, and are prepared or isolated by methods known in the art.
  • the compounds of the present invention can be used in the form of salts derived from inorganic or organic acids.
  • These salts include but are not limited to the following: acetate, adipate, alginate, citrate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, camphorate, camphorsulfonate, digluconate, cyclopentanepropionate, dodecylsulfate, ethanesulfonate, glucoheptanoate, glycerophosphate, hemisulfate, heptanoate, hexanoate, fumarate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxy-ethanesulfonate, lactate, maleate, methanesulfonate, nicotinate, 2- naphthalenesulfonate, oxalate, palmoate, pectinate, persulfate,
  • the basic nitrogen- containing groups can be quaternized with such agents as lower alkyl halides, such as methyl, ethyl, propyl, and butyl chloride, bromides, and iodides; dialkyl sulfates like dimethyl, diethyl, dibutyl, and diamyl sulfates, long chain halides such as decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides, aralkyl halides like benzyl and phenethyl bromides, and others. Water or oil-soluble or dispersible products are thereby obtained.
  • lower alkyl halides such as methyl, ethyl, propyl, and butyl chloride, bromides, and iodides
  • dialkyl sulfates like dimethyl, diethyl, dibutyl, and diamyl sulfates
  • long chain halides
  • acids which may be employed to form pharmaceutically acceptable acid addition salts include such inorganic acids as hydrochloric acid, sulphuric acid and phosphoric acid and such organic acids as oxalic acid, maleic acid, succinic acid and citric acid preferably hydrochloride salt.
  • Other examples include salts with alkali metals or alkaline earth metals, such as sodium, potassium, calcium or magnesium or with organic bases.
  • Total daily dose administered to a host in single or divided doses may be in amounts, for example, from 0.01 to 50 mg/kg body weight daily and more usually 0.1 to 30 mg. Dosage unit compositions may contain such amounts of submultiples thereof to make up the daily dose.
  • the amount of active ingredient that may be combined with the carrier materials to produce a single dosage form will vary depending upon the host treated and the particular mode of administration.
  • the dosage regimen for treating a disease condition with the compounds and/or compositions of this invention is selected in accordance with a variety of factors, including the type, age, weight, sex, diet and medical condition of the patient, the severity of the disease, the route of administration, pharmacological considerations such as the activity, efficacy, pharmacokinetic and toxicology profiles of the particular compound employed, whether a drug delivery system is utilized and whether the compound is administered as part of a drug combination.
  • the dosage regimen actually employed may vary widely and therefore may deviate from the preferred dosage regimen set forth above.
  • the compounds of the present invention may be administered orally, parenterally, by inhalation spray, rectally, or topically in dosage unit formulations containing conventional nontoxic pharmaceutically acceptable carriers, adjuvants, and vehicles as desired. Topical administration may also involve the use of transdermal administration such as transdermal patches or iontophoresis devices.
  • parenteral as used herein includes subcutaneous injections, intravenous, intramuscular, intrasternal injection, or infusion techniques.
  • sterile injectable preparations for example, sterile injectable aqueous or oleaginous suspensions may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents.
  • the sterile injectable preparation may also be a sterile injectable solution or suspension in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol.
  • acceptable vehicles and solvents that may be employed are water, Ringer's solution, and isotonic sodium chloride solution.
  • sterile, fixed oils are conventionally employed as a solvent or suspending medium.
  • any bland fixed oil may be employed including synthetic mono- or diglycerides.
  • fatty acids such as oleic acid find use in the preparation of injectables.
  • Suppositories for rectal administration of the drug can be prepared by mixing the drug with a suitable nonirritating excipient such as cocoa butter and polyethylene glycols which are solid at ordinary temperatures but liquid at the rectal temperature and will therefore melt in the rectum and release the drug.
  • Solid dosage forms for oral administration may include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound may be admixed with at least one inert diluent such as sucrose lactose or starch.
  • Such dosage forms may also comprise, as in normal practice, additional substances other than inert diluents, e.g., lubricating agents such as magnesium stearate.
  • additional substances e.g., lubricating agents such as magnesium stearate.
  • the dosage forms may also comprise buffering agents. Tablets and pills can additionally be prepared with enteric coatings.
  • Liquid dosage forms for oral administration may include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs containing inert diluents commonly used in the art, such as water.
  • Such compositions may also comprise adjuvants, such as wetting agents, emulsifying and suspending agents, and sweetening, flavoring, and perfuming agents.
  • the compounds of the invention can be administered as the sole active pharmaceutical agent, they can also be used in combination with two or three other antiviral agents which are effective against HIV- 1.
  • Such compounds include, but are not limited to; other HIV-1 protease inhibitors as disclosed in co-owned and co-filed with this application US Patent Application (Attorney Docket No. 2816) incorporated herein by reference in its entirety, various nucleoside analogs nonnucleoside reverse transcriptase inhibitors, tat antagonists and glycosidase inhibitors.
  • HIV-1 protease inhibitors include, but not limited to, Ro 31-8959 (Roberts, N.A. et al. Science 1990, 248, 358-361 and Drugs of the Future 1991, 16(3), 210-212), KNI-272, (Kagayama, S., et al. Antimicrobial Agents and Chemotherapy 1993, 810-817), the cyclic urea series (Lam, P., et al., "De Novo Design and Discovery of Potent, Nonpeptidal HIV-1 Protease Inhibitors, " paper 96 at the 205th American Chemical Society National
  • L-735,524 (Dorsey, B.D., et al., "L- 735,524: The Rational Design of a Potent and Orally Bioavailable HIV Protease Inhibitor, " paper 6 at the 206th American Chemical Society National Meeting,
  • Examples of competitive nucleoside analogs include, but are not limited to, AZT, DDI, DDC, 3TC, D4T and PMEA.
  • Examples of non-nucleoside, non-competitive reverse transcriptase inhibitors include, but are not limited to, the pyridone class (Wei, J.S., et al. J. Med. Chem. 1993, 36, 249-255; Hoffman, J.M., et al. J. Med. Chem. 1992, 35, 3784-3791; Saari et al. J. Med. Chem. 1992, 35 3792-3802; Drugs of the Future 1992, 17(4), 283-285, and analogs thereof); the bis-
  • heteroaryl piperazines class (heteroaryl)piperazines class (Romero, D.L., et al. J. Med. Chem. 1993, 36, 1505-1508; Romero, D.L., et al. Proc. Natl. Acad. Sci. USA 1991, 34, 746-751 and 3187- 3198; and analogs thereof) and the tricyclic pyridobenzo- and depyridodiazepinones (Hargrave, K.D., J. Med. Chem. 1991, 34, 2231-2241; Merluzzi, M.J.
  • tat antagonists include, but are not limited to, Ro 5-3335 and Ro 24-7429 (Hsu, M.C. et al., Proc. Natl. Acad. Sci. USA 1993, 909, 6395-6399; Tarn, S.
  • glycosidase inhibitors include, but are not limited to, castanospermine, castanospermine 6-butryl ester, N-butyl-1-deoxynojirimycin, N-butyl-1- deoxynojiri ycin per-butryl ester and analogs and prodrugs thereof.
  • the therapeutic agents can be formulated as separate compositions which are given at the same time or different times, or the therapeutic agents can be given as a single composition.
  • the compounds of the present invention are effective antiviral compounds and, in particular, are effective retroviral inhibitors as shown above.
  • the subject compounds are effective HIV protease inhibitors. It is contemplated that the subject compounds will also inhibit other retroviruses such as other lentiviruses in particular other strains of HIV, e.g. HIV-2, human T-cell leukemia virus, respiratory syncitial virus, simia immunodeficiency virus, feline leukemia virus, feline immuno-deficiency virus, hepadnavirus, cytomegalovirus and picornavirus.
  • the subject compounds are effective in the treatment and/or proplylaxis of retroviral infections.
  • the subject compounds are also effective in preventing the growth of retroviruses in a solution.
  • Both human and animal cell cultures such as T- lymphocyte cultures, are utilized for a variety of well known purposes, such as research and diagnostic procedures including calibrators and controls.
  • the subject compounds Prior to and during the growth and storage of a cell culture, the subject compounds may be added to the cell culture medium at an effective concentration to prevent the unexpected or undesired replication of a retrovirus that may inadvertently or unknowingly be present in the cell culture.
  • the virus may be present originally in the cell culture, for example HIV is known to be present in human T-lymphocytes long before it is detectable in blood, or through exposure to the virus. This use of the subject compounds prevents the unknowing or inadvertent exposure of a potentially lethal retrovirus to a researcher or clinician.

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Abstract

L'invention concerne la préparation et l'utilisation d'inhibiteurs de protéases rétrovirales représentés par la formule (I) ainsi que leurs combinaisons, lesquels sont efficaces dans la prévention de la réplication de rétrovirus mammifères, tel que le virus de l'immunodéficience humaine (VIH).
PCT/US1994/008697 1993-08-20 1994-08-09 Inhibiteurs de proteases retrovirales et leurs combinaisons WO1995006061A1 (fr)

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Cited By (12)

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WO1998035685A1 (fr) * 1997-02-14 1998-08-20 G.D. Searle And Co. Utilisation de composes de n-substitue-1,5-didesoxy-1,5-imino-d-glucitol dans des traitements associes contre des infections provoquees par des virus de l'hepatite
US6225325B1 (en) 1997-11-10 2001-05-01 G.D. Searle & Company Use of alkylated iminosugars to treat multidrug resistance
US6232333B1 (en) 1996-11-21 2001-05-15 Abbott Laboratories Pharmaceutical composition
US6515028B1 (en) 1999-02-12 2003-02-04 G.D. Searle & Co. Glucamine compounds for treating hepatitis virus infections
US6545021B1 (en) 1999-02-12 2003-04-08 G.D. Searle & Co. Use of substituted-1,5-dideoxy-1,5-imino-D-glucitol compounds for treating hepatitis virus infections
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US6809083B1 (en) 1998-02-12 2004-10-26 Richard A. Mueller Use of N-substituted-1, 5-dideoxy-1, 5-imino-D-glucitol compounds for treating hepatitis virus infections
EP1917958A2 (fr) 2000-01-19 2008-05-07 Abbott Laboratories Formules pharmaceutiques améliorées d' inhibiteurs de la HIV protease
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WO2010144869A2 (fr) 2009-06-12 2010-12-16 Nektar Therapeutics Inhibiteurs de protéase
EP2522367A1 (fr) 2007-03-12 2012-11-14 Nektar Therapeutics Conjugués oligomer-protéase inhibiteurs
US9095620B2 (en) 2008-03-12 2015-08-04 Nektar Therapeutics Reagents

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US6458818B1 (en) 1996-11-21 2002-10-01 Abbott Laboratories Pharmaceutical composition
US6521651B1 (en) 1996-11-21 2003-02-18 Abbott Laboratories Pharmaceutical composition
WO1998035685A1 (fr) * 1997-02-14 1998-08-20 G.D. Searle And Co. Utilisation de composes de n-substitue-1,5-didesoxy-1,5-imino-d-glucitol dans des traitements associes contre des infections provoquees par des virus de l'hepatite
US7612093B2 (en) 1997-02-14 2009-11-03 United Therapeutics Corporation Compositions of treating hepatitis virus infections with N-substituted-1,5-dideoxy-1,5-imino-D-glucitol compounds in combination therapy
CN100382804C (zh) * 1997-02-14 2008-04-23 G·D·瑟尔公司 N-取代的-1,5-二脱氧-1,5-亚氨基-d-山梨醇化合物在制备治疗肝炎病毒感染的药物中的用途
US6225325B1 (en) 1997-11-10 2001-05-01 G.D. Searle & Company Use of alkylated iminosugars to treat multidrug resistance
US6809083B1 (en) 1998-02-12 2004-10-26 Richard A. Mueller Use of N-substituted-1, 5-dideoxy-1, 5-imino-D-glucitol compounds for treating hepatitis virus infections
US6689759B1 (en) 1998-02-12 2004-02-10 G. D. Searle & Co. Methods of Treating hepatitis virus infections with N-substituted-1,5-dideoxy-1,5-imino-d-glucitol compounds in combination therapy
US6515028B1 (en) 1999-02-12 2003-02-04 G.D. Searle & Co. Glucamine compounds for treating hepatitis virus infections
US6747149B2 (en) 1999-02-12 2004-06-08 G. D. Searle & Co. Glucamine salts for treating hepatitis virus infections
US6545021B1 (en) 1999-02-12 2003-04-08 G.D. Searle & Co. Use of substituted-1,5-dideoxy-1,5-imino-D-glucitol compounds for treating hepatitis virus infections
EP1917958A2 (fr) 2000-01-19 2008-05-07 Abbott Laboratories Formules pharmaceutiques améliorées d' inhibiteurs de la HIV protease
EP2269591A2 (fr) 2000-01-19 2011-01-05 Abbott Laboratories Compositions pharmaceutiques ameliorées
EP2522367A1 (fr) 2007-03-12 2012-11-14 Nektar Therapeutics Conjugués oligomer-protéase inhibiteurs
US8598364B2 (en) 2007-03-12 2013-12-03 Nektar Therapeutics Oligomer-protease inhibitor conjugates
US9107956B2 (en) 2007-03-12 2015-08-18 Nektar Therapeutics Oligomer-protease inhibitor conjugates
US9095620B2 (en) 2008-03-12 2015-08-04 Nektar Therapeutics Reagents
WO2010144869A2 (fr) 2009-06-12 2010-12-16 Nektar Therapeutics Inhibiteurs de protéase

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AU7518694A (en) 1995-03-21
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