RELATED APPLICATION

This application claims benefit of U.S. Serial No. 61/347141 , filed May 21 , 2010, herein incorporated by reference.

FIELD OF THE INVENTION

The present invention relates to dihydropyrrolopyrazole compounds and their use as inhibitors of human immunodeficiency virus (HIV) capsid disassembly,

pharmaceutical compositions containing such compounds and methods for using these compounds in the treatment of HIV infection.

BACKGROUND OF THE INVENTION

To date, over 20 FDA-approved drugs make up the antiretroviral arsenal against HIV (1). These compounds generally target the viral enzymes or the viral entry process and can be divided within 6 mechanistic classes. The standard of care is a multi-drug therapeutic regime, often referred to as highly active antiretroviral therapy (HAART), where combinations of at least three drugs targeting the virus are administered. Although HAART can successfully restrict viral replication for many years, drug resistance can still occur. Cross-resistance within mechanistic classes and the emergence of multi-drug-resistant (MDR) isolates can have considerable impact on treatment options and patient outcome. This underlines the need to discover novel classes of HIV inhibitors.

The HIV-1 Capsid protein (CA), which plays an essential role in the viral replication cycle, may represent such a novel therapeutic target (2). CA is a domain of the GAG polyprotein, where it contributes some of the key protein-protein interactions required for the assembly of immature viral particles. During viral maturation, proteolytic cleavage of GAG releases CA which re-assembles to form a cone shaped structure called the core, enclosing the viral RNA genome and enzymatic activities required for infectivity. Core formation is driven by a multitude of weak protein/protein interactions (2). CA mutations that prevent core assembly result in non-infectious viral particles (3-5). CA-binding inhibitors of capsid assembly have been reported previously (6, 7), providing evidence that CA may be a viable drug target. Interestingly, the only inhibitors for which structural information is currently available seem to target the NTD-CTD interface, suggesting it may represent an inhibition 'hot-spot' (7, 8). References- (1 ) International Journal of Antimicrobial Agents 2009, 33(4), 307-320. (2) Current Opinion in Structural Biology 2008, 18, 203-17. (3) Journal of Virology 2002, 76, 5667-5677. (4) Journal of Virology 2004, 78, 2545-2552. (5) Journal of Virology 2003, 77, 5439-50. (6) Journal of Molecular Biology 2003, 327, 1013-20. (7) Λ/aftvre Structural and Molecular Biology 2005, 72, 678-82. (8) Journal of Molecular Biology 2007, 373, 355-66.

WO 2008/120725 discloses pyrrolinone derivatives as P2X3 and/or P2X2/3 antagonists.

SUMMARY OF THE INVENTION

The present invention provides a novel series of compounds having inhibitory activity against HIV replication. The compounds of the present invention have inhibitory activity against HIV-1 capsid disassembly. Further objects of this invention arise for the one skilled in the art from the following description and the examples.

One aspect of the invention provides compounds of formula (I) and an isomer, tautomer, racemate, enantiomer or diastereomer thereof:

wherein R is aryl or Het,

wherein each said aryl or Het is optionally substituted 1 to 3 times with halo, -(C ₁ _ ₆ )haloalkyl, -0-(C ₁ _ ₆ )haloalkyl, OH, COOH, -C(=0)-0-(C ₁ _ ₆ )alkyl, -C(=0)- NH-(C ₁ _ ₆ )alkyl, N0 ₂ , R ¹¹ , -O-R ¹¹ , NH ₂ , -NH((C ₁ _ ₆ )alkyl), -N((C ₁ _ ₆ )alkyl) ₂ , -

NH((C ₁ _ ₆ )alkyl-aryl), -NH-C(=0)-NH-aryl, -NH-S0 ₂ -R ¹¹ or -NH-S0 ₂ -((C ₂ . ₆ )alkenyl-aryl);

_ ₆ )alkyl, -(C-i. ₆ )alkyl-aryl, -(C-i. ₆ )alkyl-Het, aryl or Het, wherein each said alkyl, aryl and Het, either alone or in combination with another radical, are optionally substituted 1 to 3 times with COOH, -C(=0)-0- (d_ ₆ )alkyl, -C(=0)-NH-(C ₁ _ ₆ )alkyl, halo, OH, -0-(C ₁ _ ₆ )alkyl, N0 ₂ , NH ₂ , - NH(d_6)alkyl), -N((C ₁ _ ₆ )alkyl) ₂ , -NH-C(=0)-(d_ ₆ )alkyl, -NH-C(=0)-0-(d_ 6)alkyl, -C(=0)-NH ₂ , -(C ₁ . ₆ )alkyl-NH-C(=0)-(C ₁ . ₆ )alkyl, azido or Het optionally substituted 1 to 2 times with (C-i_ ₆ )alkyl;

R ² is halo, -(d-e)alkyl-aryl, -(C-i. ₆ )alkyl-Het, -NH-(d_ ₆ )alkyl-aryl, -0-(C-i. ₆ )alkyl-aryl, - 0-(d- ₆ )alkenyl-aryl or -0-(d_ ₆ )alkyl-Het,

wherein each said aryl and Het, either alone or in combination with another radical, are optionally substituted 1 to 3 times with halo, -0-(C-i_ ₆ )alkyl, -(d-

₆ )alkyl, -(C ₂ - ₆ )alkenyl, -(d_ ₆ )haloalkyl, OH or azido; n is 0, 1 or 2;

R ³ is H or (C ₁ _ ₆ )alkyl optionally substituted with -N((C _ ₆ )alkyl) ₂ , -C(=0)

OH or -C(=0)-0-(d_ ₆ )alkyl;

R is H or (d_ ₆ )alkyl;

_ ₇ )cycloalkyl, -(C-i_ ₆ )alkyl-aryl, -(C-i_ ₆ )alkyl-Het, aryl or Het;

wherein each said (C ₃ . ₇ )cycloalkyl, aryl and Het, either alone or in combination with another radical, are optionally substituted 1 to 3 times with halo, -0-(d_ ₆ )alkyl, -(d_ ₆ )alkyl, -(d_ ₆ )haloalkyl, OH, COOH, -C(=0)-0-(d_ 6)alkyl, -C(=0)NH ₂ , NH ₂ , -NH(d_ ₆ )alkyl), -N((d_ ₆ )alkyl) ₂ , -S0 ₂ -(d_ ₆ )alkyl, - (C-i- ₆ )alkyl-aryl or -(Ci_ ₆ )alkyl-Het; and

wherein said alkyl portion of alkyl-aryl and alkyl-Het is optionally substituted 1 to 2 times with -C(=0)NH ₂ or -C(=0)-0-(d_ ₆ )alkyl; or

R ³¹ and R ³² , together with the N to which they are attached, are linked to form a 4- to 10-membered Het optionally further containing 1 to 3 heteroatoms each

independently selected from the group consisting of N, O and S;

wherein said heterocycle is optionally substituted 1 to 3 times with halo, -0-(C-i_ ₆ )alkyl, -(d. ₆ )alkyl optionally substituted with -C(=0)-0- (C _1-6 )alkyl, -(C ₁ _ ₆ )haloalkyl, OH, COOH, -C(=0)NH ₂ , -C(=0)-NH-(d_ 6)alkyl, N0 ₂ , NH ₂ , -NH(d_6)alkyl), -N((C ₁ _ ₆ )alkyl) ₂ , -(C ₁ _ ₆ )alkyl-aryl, - (C-i. ₆ )alkyl-Het, aryl optionally substituted with -(C _ ₆ )alkyl, -0-(C-|. 6)alkyl or N0 ₂ , or Het optionally substituted with halo, -(Ci_ ₆ )alkyl or - (C _1-6 )haloalkyl; or a salt thereof; with the proviso that the following compounds are excluded:

4,5-dihydro-3,4-diphenyl-pyrrolo[3,4-c]pyrazol-6(1 H)-one;

4-(4-chlorophenyl)-4,5-dihydro-3-phenyl-pyrrolo[3,4-c]pyrazo l-6(1 H)-one;

4-(4-hydroxyphenyl)-4,5-dihydro-3-phenyl-pyrrolo[3,4-c]pyraz ol-6(1 H)-one;

4-(2-fluorophenyl)-4,5-dihydro-3-phenyl-pyrrolo[3,4-c]pyrazo l-6(1 H)-one;

4-(4-ethylphenyl)-4,5-dihydro-3-phenyl-pyrrolo[3,4-c]pyrazol -6(1 H)-one; and 4,5-dihydro-3-phenyl-4-(3-pyridinyl)-pyrrolo[3,4-c]pyrazol-6 (1 H)-one.

Another aspect of this invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, as a medicament. Included within the scope of this invention is a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof, in admixture with at least one

pharmaceutically acceptable carrier medium or auxiliary agent. According to a further aspect of this invention, the pharmaceutical composition according to this invention further comprises a therapeutically effective amount of at least one other antiviral agent.

The invention also provides the use of a pharmaceutical composition as described hereinabove for the treatment of an HIV infection in a human being having or at risk of having the infection.

Another important aspect of the invention involves a method of treating or preventing an HIV infection in a human being by administering to the human being a therapeutically effective amount of a compound of formula (I), a pharmaceutically acceptable salt thereof, or a composition as described above, alone or in combination with at least one other antiviral agent, administered together or separately.

Also within the scope of this invention is the use of a compound of formula (I), or a pharmaceutically acceptable salt thereof, as described herein, for the manufacture of a medicament for the treatment or prevention of an HIV infection in a human being.

A further aspect of the invention is a compound of formula (I) or a pharmaceutically acceptable salt thereof for treatment or prevention of HIV infection.

Still another aspect of this invention relates to a method of inhibiting the replication of HIV comprising exposing the virus to an effective amount of the compound of formula (I), or a salt thereof, under conditions where replication of HIV is inhibited.

Further included in the scope of the invention is the use of a compound of formula (I), or a salt thereof, to inhibit the replication of HIV.

DETAILED DESCRIPTION OF THE INVENTION DEFINITIONS

Terms not specifically defined herein should be given the meanings that would be given to them by one of skill in the art in light of the disclosure and the context. As used in the specification, however, unless specified to the contrary, the following terms have the meaning indicated and the following conventions are adhered to. In the groups, radicals, or moieties defined below, the number of carbon atoms is often specified preceding the group, for example, Ci_ ₆ -alkyl means an alkyl group or radical having 1 to 6 carbon atoms. In general, for groups comprising two or more subgroups, the first named subgroup is the radical attachment point, for example, the substituent "-C-i. ₃ -alkyl-aryl" means an aryl group which is bound to a -C-i_ ₃ -alkyl group, wherein the -C-i_ ₃ -alkyl-group is bound to the core. In the previous example of "-C-i. ₃ -alkyl-aryl", substituents may be attached to either the C _ ₃ -alkyl or aryl portion thereof or both, unless specified otherwise. The term "C _ _n -alkyl", wherein n is an integer from 2 to n, either alone or in combination with another radical denotes an acyclic, saturated, branched or linear hydrocarbon radical with 1 to n C atoms. For example the term C _ ₅ -alkyl includes, but is not limited to, the radicals H ₃ C-, H ₃ C-CH ₂ -, H ₃ C-CH ₂ -CH ₂ -, H ₃ C-CH(CH ₃ )-, H ₃ C-CH ₂ -CH(CH ₃ )-, H ₃ C-CH(CH ₃ )-CH ₂ -, H ₃ C-C(CH ₃ ) ₂ -, H ₃ C-CH ₂ -CH ₂ -CH(CH ₃ )-, H ₃ C-CH(CH ₃ )-CH ₂ -CH ₂ -, H ₃ C-CH ₂ -C(CH ₃ ) ₂ - and H ₃ C-C(CH ₃ ) ₂ -CH ₂ -.

The term "C ₂ . _n -alkenyl", is used for a group as defined in the definition for with at least two carbon atoms, if at least two of those carbon atoms of said group are bonded to each other by a double bond.

The term "carbocycle" means a mono- or multi-ring ring structure consisting only of carbon containing between one and four rings wherein such rings may be attached together in a pendent manner or may be fused. The term "carbocycle" refers to fully saturated and aromatic ring systems and partially saturated ring systems. The term "carbocycle" additionally encompasses spiro systems, and bridged systems.

The term "C ₃ . _n -cycloalkyl", wherein n is an integer 4 to n, either alone or in combination with another radical denotes a cyclic, saturated, unbranched hydrocarbon radical with 3 to n C atoms. For example the term C ₃ . ₇ -cycloalkyl includes cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl.

The term "halo" generally denotes fluorine, chlorine, bromine and iodine.

The term "azido" generally denotes the group -N ₃ (-N=N ⁺ =N-).

The term "aryl" as used herein, either alone or in combination with another radical, denotes a carbocyclic aromatic monocyclic group containing 6 carbon atoms which may be further fused one or more 5- or 6-membered carbocyclic group which may be aromatic, saturated or unsaturated. Aryl includes, but is not limited to, phenyl, indanyl, indenyl, naphthyl, anthracenyl, phenanthrenyl, tetrahydronaphthyl and dihydronaphthyl.

The term "Het" as used herein, either alone or in combination with another radical, denotes a heterocyclyl or heteroaryl ring system. The term "heterocyclyl" means a saturated or unsaturated mono- or polycyclic-ring systems including aromatic ring system containing one or more heteroatoms selected from N, O or S(0) _r with r=0, 1 or 2 wherein none of the heteroatoms is part of the aromatic ring. The term "heterocyclyl" is intended to include all the possible isomeric forms. Thus, the term "heterocyclyl" includes the following exemplary structures which are not depicted as radicals as each form may be attached through d:

The term "heteroaryl" means a mono- or polycyclic-ring systems containing one or more heteroatoms selected from N, O or S(0) _r with r=0, 1 or 2 wherein at least one of the heteroatoms is part of aromatic ring. The term "heteroaryl" is intended to include all the possible isomeric forms. Thus, the term "heteroaryl" includes the following exemplary structures which are not depicted as radicals as each form may be attached through a covalent bond to any atom so long as appropriate valences are maintained:

Many of the terms given above may be used repeatedly in the definition of a formula or group and in each case have one of the meanings given above, independently of one another.

In case a compound of the present invention or an intermediate used in the synthesis of a compound of the present invention is depicted in the form of a chemical name and as a formula, the formula shall prevail in case of any discrepancy between the name and formula.

An asterisk or the designation ' is used in sub-formulas to indicate the bond which is connected to the core molecule as defined.

As used herein, the designation whereby a bond to a substituent R is drawn as emanatin from the center of a ring system, such as, for example, , is intended to mean that the substituent R may be attached to any free position on the ring system that would otherwise be substituted with a hydrogen atom, unless specified otherwise.

Unless specifically indicated, throughout the specification and the appended claims, a given chemical formula or name shall encompass tautomers and all stereo, optical and geometrical isomers (e.g. enantiomers, diastereomers, E/Z isomers, atropisomers) and racemates thereof as well as mixtures in different proportions of the separate enantiomers, mixtures of diastereomers, or mixtures of any of the foregoing forms where such isomers and enantiomers exist, as well as salts, including pharmaceutically acceptable salts thereof and solvates thereof such as for instance hydrates including solvates of the free compounds or solvates of a salt of the compound.

One skilled in the art would know how to separate, enrich, or selectively prepare the enantiomers of the compounds of the present invention. Preparation of pure stereoisomers, e.g. enantiomers and diastereomers, or mixtures of desired enantiomeric excess (ee) or enantiomeric purity, are accomplished by one or more of the many methods of (a) separation or resolution of enantiomers, or (b) enantioselective synthesis known to those of skill in the art, or a combination thereof. These resolution methods generally rely on chiral recognition and include but not limited to chromatography using chiral stationary phases, enantioselective host-guest complexation, resolution or synthesis using chiral auxiliaries, enantioselective synthesis, enzymatic and nonenzymatic kinetic resolution, or spontaneous enantioselective crystallization. Such methods are disclosed generally in Chiral Separation Techniques: A Practical Approach (2nd Ed.), G. Subramanian (ed.), Wiley-VCH, 2000; T.E. Beesley and R.P.W. Scott, Chiral Chromatography, John Wiley & Sons, 1999; and Satinder Ahuja, Chiral Separations by

Chromatography, Am. Chem. Soc, 2000. Furthermore, there are equally well-known methods for the quantitation of enantiomeric excess or purity, including but not limited to GC, HPLC, CE, or NMR, and assignment of absolute configuration and conformation, including but not limited to CD, ORD, X-ray crystallography, or NMR.

The phrase "pharmaceutically acceptable" is employed herein to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, and commensurate with a reasonable benefit/risk ratio. As used herein, "pharmaceutically acceptable salts" refer to derivatives of the disclosed compounds wherein the parent compound is modified by making acid or base salts thereof. 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. For example, such salts include salts from ammonia, L-arginine, betaine, benethamine, benzathine, calcium hydroxide, choline, deanol, diethanolamine (2,2'- iminobis(ethanol)), diethylamine, 2-(diethylamino)-ethanol, 2-aminoethanol, ethylenediamine, N-ethyl-glucamine, hydrabamine, 1 H-imidazole, lysine, magnesium hydroxide, 4-(2-hydroxyethyl)-morpholine, piperazine, potassium hydroxide, 1-(2-hydroxyethyl)-pyrrolidine, sodium hydroxide, triethanolamine (2,2',2"-nitrilotris(ethanol)), tromethamine, zinc hydroxide, acetic acid, 2.2-dichloro- acetic acid, adipic acid, alginic acid, ascorbic acid, L-aspartic acid, benzenesulfonic acid, benzoic acid, 2,5-dihydroxybenzoic acid, 4-acetamido-benzoic acid, (+)- camphoric acid, (+)-camphor-10-sulfonic acid, carbonic acid, cinnamic acid, citric acid, cyclamic acid, decanoic acid, dodecylsulfuric acid, ethane-1 ,2-disulfonic acid, ethanesulfonic acid, 2-hydroxy-ethanesulfonic acid, ethylenediaminetetraacetic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, D-glucoheptonic acid, D- gluconic acid, D-glucuronic acid, glutamic acid, , glutaric acid, 2-oxo-glutaric acid, glycerophosphoric acid, glycine, glycolic acid, hexanoic acid, hippuric acid, hydrobromic acid, hydrochloric acid, isobutyric acid, DL-lactic acid, lactobionic acid, lauric acid, lysine, maleic acid, (-)-L-malic acid, malonic acid, DL-mandelic acid, methanesulfonic acid, galactaric acid, naphthalene-1 ,5-disulfonic acid, naphthalene- 2-sulfonic acid, 1 -hydro xy-2-naphthoic acid, nicotinic acid, nitric acid, octanoic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid (embonic acid), phosphoric acid, propionic acid, (-)-L-pyroglutamic acid, salicylic acid, ^aminosalicylic acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, tannic acid, (+)- L-tartaric acid, thiocyanic acid, p-toluenesulfonic acid and undecylenic acid. Further pharmaceutically acceptable salts can be formed with cations from metals like aluminium, calcium, lithium, magnesium, potassium, sodium, zinc and the like, (also see Pharmaceutical salts, Berge, S.M. et al., J. Pharm. Sci., (1977), 66, 1-19, incorporated herein by reference).

The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound which contains 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 sufficient amount of the appropriate base or acid in water or in an organic diluent like ether, ethyl acetate, ethanol, isopropanol, or acetonitrile, or a mixture thereof.

Salts of other acids than those mentioned above which for example are useful for purifying or isolating the compounds of the present invention (e.g. trifluoro acetate salts) also comprise a part of the invention.

The term "treatment" as used herein is intended to mean the administration of a compound or composition according to the present invention to alleviate or eliminate symptoms of HIV infection and/or to reduce viral load in a patient. The term

"treatment" also encompasses the administration of a compound or composition according to the present invention post-exposure of the individual to the virus but before the appearance of symptoms of the disease, and/or prior to the detection of the virus in the blood, to prevent the appearance of symptoms of the disease and/or to prevent the virus from reaching detectable levels in the blood, and the administration of a compound or composition according to the present invention to prevent perinatal transmission of HIV-1 from mother to baby, by administration to the mother before giving birth and to the child within the first days of life.

The term "antiviral agent" as used herein is intended to mean an agent that is effective to inhibit the formation and/or replication of a virus in a human being, including but not limited to agents that interfere with either host or viral mechanisms necessary for the formation and/or replication of a virus in a human being.

Preferred embodiments

In the following preferred embodiments, groups and substituents of the compounds of Formula (I) according to this invention are described in detail.

Any and each of the following definitions R ¹ -A to R ¹ -C, R ² -A to R ² -C, R ³ -A to R ³ -C and n-A to n-C may be combined with one another.

El

R ¹ -A: R ¹ is aryl or Het,

wherein each said aryl or Het is optionally substituted 1 to 3 times with halo, -(C ₁ _ ₆ )haloalkyl, -0-(C ₁ _ ₆ )haloalkyl, OH, COOH, -C(=0)-0-(C ₁ _ ₆ )alkyl, -C(=0)- NH-(d_ ₆ )alkyl, N0 ₂ , R ¹¹ , -O-R ¹¹ , NH ₂ , -NH((d_ ₆ )alkyl), -N((d_ ₆ )alkyl) ₂ , - NH((C ₁ _ ₆ )alkyl-aryl), -NH-C(=0)-NH-aryl, -NH-C(=0)-(d_ ₆ )alkyl, -NH-S0 ₂ -R ¹¹ or -NH-S0 ₂ -((C ₂ . ₆ )alkenyl-aryl);

R ¹¹ is -(C-i. ₆ )alkyl, -(C-i_ ₆ )alkyl-aryl, -(C-i. ₆ )alkyl-Het, aryl or Het,

wherein each said alkyl, aryl and Het, either alone or in combination with another radical, are optionally substituted 1 to 3 times with COOH, -C(=0)-0-(C ₁ _ ₆ )alkyl, -C(=0)-NH-(C ₁ _ ₆ )alkyl, halo, OH, -0-(d_ 6)alkyl, N0 ₂ , NH ₂ , -NH(d_ ₆ )alkyl), -N((d_ ₆ )alkyl) ₂ , -NH-C(=0)-(d_ 6)alkyl, -NH-C(=0)-0-(d_ ₆ )alkyl, -C(=0)-NH ₂ , -(d_ ₆ )alkyl-NH-C(=0)- (C _ ₆ )alkyl, azido or Het optionally substituted 1 to 2 times with (d_ 6)alkyl.

R ¹ -B: R ¹ is phenyl or Het,

wherein each said phenyl or Het is substituted 1 to 2 times with -0-(d_ 6)haloalkyl, OH, COOH, -C(=0)-0-(d_ ₃ )alkyl, -C(=0)-NH-(d_ ₃ )alkyl, N0 ₂ , R ¹¹ , -O-R ¹¹ , NH ₂ , -NH((d_ ₃ )alkyl), -N((d_ ₃ )alkyl) ₂ , -NH((d_ ₃ )alkyl- phenyl), - NH-C(=0)-NH- phenyl, -NH-C(=0)-(d_ ₃ )alkyl, -NH-S0 ₂ -R ¹¹ or -NH-S0 ₂ -((C ₂ _ 4)alkenyl-phenyl);

R ¹¹ is -(C _ ₃ )alkyl, -(d_ ₃ )alkyl-phenyl, -(d_ ₃ )alkyl-Het, phenyl or Het,

wherein each said phenyl and Het, either alone or in combination with another radical, are optionally substituted 1 to 2 times with COOH, -C(=0)-0-(d_ ₃ )alkyl, -C(=0)-NH-(d_ ₃ )alkyl, halo, OH, -0-(d 3)alkyl, N0 ₂ , NH ₂ , -NH(C ₁ _ ₃ )alkyl), -N((C ₁ _ ₃ )alkyl) ₂ , -NH-C(=0)-(d_ 3)alkyl, -NH-C(=0)-0-(d_ ₃ )alkyl, -C(=0)-NH ₂ , -(d_ ₃ )alkyl-NH-C(=0)- (d- ₃ )alkyl, azido or Het optionally substituted 1 to 2 times with (d_ 3)alkyl; and

Het is defined as a 5- or 6-membered heteroaryl or heterocyclyl ring system.

R ¹ is phenyl or pyridyl,

wherein each said phenyl or pyridyl is substituted 1 to 2 times with -0-(d_ 3)haloalkyl, OH, COOH, -C(=0)-0-(d_ ₃ )alkyl, -C(=0)-NH-(d_ ₃ )alkyl, R ¹¹ , -O- R ¹¹ , NH ₂ , -NH((d_ ₃ )alkyl), -N((d_ ₃ )alkyl) ₂ , -NH((d_ ₃ )alkyl-phenyl) or -NH- C(=0)-(d. ₃ )alkyl;

R ¹¹ is -(C-i_ ₃ )alkyl, -(d_ ₃ )alkyl-phenyl, -(d_ ₃ )alkyl-Het, phenyl or Het,

wherein each said alkyl, phenyl and Het, either alone or in combination with another radical, are optionally substituted 1 to 2 times with COOH, -C(=0)-0-(d_ ₃ )alkyl, -C(=0)-NH-(d_ ₃ )alkyl, halo, OH, -0-(d_ ₃ )alkyl, NH ₂ , -NH(d_ ₃ )alkyl), -N((d_ ₃ )alkyl) ₂ , -NH-C(=0)- (d_ ₃ )alkyl, -NH-C(=0)-0-(d_ ₃ )alkyl, -C(=0)-NH ₂ , -(d_ ₃ )alkyl-NH- C(=0)-(d_ ₃ )alkyl or Het optionally substituted 1 to 2 times with (d_ 3)alkyl; and

Het is defined as a 5- or 6-membered heteroaryl or heterocyclyl ring system. R ²

R ² -A: R ² is halo, -(d_ ₆ )alkyl-aryl, -(d_ ₆ )alkyl-Het, -NH-(d_ ₆ )alkyl-aryl, -0-(d_ ₆ )alkyl- aryl, -0-(C ₂ . ₆ )alkenyl-aryl or -0-(C-i. ₆ )alkyl-Het,

wherein each said aryl and Het, either alone or in combination with another radical, are optionally substituted 1 to 3 times with halo, -O- (d_ ₆ )alkyl, -(d_ ₆ )alkyl, -(C ₂ . ₆ )alkenyl, -(d_ ₆ )haloalkyl, OH or azido.

R ² -B: R ² is -(d_ ₃ )alkyl-phenyl, -(d_ ₃ )alkyl-Het, -0-(d_ ₃ )alkyl-phenyl, -0-(C ₂ .

4)alkenyl-phenyl or -0-(d_ ₃ )alkyl-Het,

wherein each said phenyl and Het, either alone or in combination with another radical, are optionally substituted 1 to 2 times with halo, -0-(C _1-3 )alkyl, -(C _1-3 )alkyl, -(C ₂ - ₄ )alkenyl, -(C _1-3 )haloalkyl, OH or azido; and

Het is defined as a 5- or 6-membered heteroaryl or heterocyclyl ring system.

R ² -C: R ² is -0-(C _1-3 )alkyl-phenyl or -0-(C _1-3 )alkyl-Het,

wherein each said phenyl and Het, either alone or in combination with another radical, are optionally substituted 1 to 2 times with halo, -0-(C _1-3 )alkyl, -(C _1-3 )alkyl, -(C _2- )alkenyl, -(C _1-3 )haloalkyl, OH or azido; and

Het is defined as a 5- or 6-membered heteroaryl or heterocyclyl ring system.

n

n-A n is 0, 1 or 2.

n-B n is 0 or 1 .

n-C n is 0.

R ³ is H or (C _1-6 )alkyl optionally substituted with -N((C _1-6 )alkyl) ₂ , -C(=0)- N(R ³¹ )(R ³² ), OH or -C(=0)-0-(C ₁ _ ₆ )alkyl);

R ³¹ is H or (C ₁ _ ₆ )alkyl;

R ³² is (C _3-7 )cycloalkyl, -(C _1-6 )alkyl-aryl, -(C _1-6 )alkyl-Het, aryl or Het;

wherein each said (C _3-7 )cycloalkyl, aryl and Het, either alone or in combination with another radical, are optionally substituted 1 to 3 times with halo, -0-(C _1-6 )alkyl, -(C _1-6 )alkyl, -(C _1-6 )haloalkyl, OH, COOH, -C(=0)-0-(C ₁ _ ₆ )alkyl, -C(=0)NH ₂ , NH ₂ , -NH(C ₁ _ ₆ )alkyl), - N((Ci _-6 )alkyl) ₂ , -S0 ₂ -(Ci _-6 )alkyl, -(C _ ₆ )alkyl-aryl or -(Ci _-6 )alkyl-Het; and wherein said alkyl portion of alkyl-aryl and alkyl-Het is optionally substituted 1 to 2 times with -C(=0)NH ₂ or -C(=0)-0-(Ci _-6 )alkyl; or R ³¹ and R ³² , together with the N to which they are attached, are linked to form a 4- to 10-membered Het optionally further containing 1 to 3 heteroatoms each independently selected from the group consisting of N, O and S;

wherein said Het is optionally substituted 1 to 3 times with halo, -O- (C _1-6 )alkyl, -(C _1-6 )alkyl optionally substituted with -C(=0)-0-(C _1-6 )alkyl, -(C ₁ _ ₆ )haloalkyl, OH, COOH, -C(=0)NH ₂ , -C(=0)-NH-(C _1-6 )alkyl, N0 ₂ , NH ₂ , -NH(d_ ₆ )alkyl), -N((C ₁ _ ₆ )alkyl) ₂ , -(C ₁ _ ₆ )alkyl-aryl, -(d_ ₆ )alkyl-Het, aryl optionally substituted with -(C _ ₆ )alkyl, -0-(C-i. ₆ )alkyl or N0 ₂ , or Het optionally substituted with halo, -(C _ ₆ )alkyl or -(d. ₆ )haloalkyl. R ³ -B: R ³ is H or (C ₁ _ ₃ )alkyl substituted with -N((C ₁ _ ₃ )alkyl) ₂ or -C(=0)-N(R ³¹ )(R ³² );

R ³¹ is H or (C ₁ _ ₃ )alkyl;

R ³² is -(C-i. ₃ )alkyl-phenyl, -(C-i. ₃ )alkyl-Het, phenyl or Het;

wherein Het is defined as a 5- or 6-membered heteroaryl or heterocyclyl ring system;

wherein each said phenyl and Het, either alone or in combination with another radical, are optionally substituted 1 to 2 times with halo,

-0-(d_ ₃ )alkyl, -(d_ ₃ )alkyl, -(d_ ₃ )haloalkyl, OH, COOH, -C(=0)-0-(d_

₃ )alkyl, -C(=0)NH ₂ , NH ₂ , -NH(d_ ₃ )alkyl), -N((d_ ₃ )alkyl) ₂ , -S0 ₂ -(d_

₃ )alkyl or -(C-i_ ₃ )alkyl-phenyl; and

wherein said alkyl portion of alkyl-phenyl and alkyl-Het is optionally substituted 1 to 2 times with -C(=0)NH ₂ or -C(=0)-0-(d_ ₃ )alkyl; or R ³¹ and R ³² , together with the N to which they are attached, are linked to form a 5-, 6-, 9- or 10-membered Het optionally further containing 1 to 2 N heteroatoms;

wherein said Het is optionally substituted 1 to 2 times with halo, -O-

(C-i_ ₃ )alkyl, -(C _ ₃ )alkyl optionally substituted with -C(=0)-0-(d_ ₃ )alkyl, -(d_ ₃ )haloalkyl, OH, COOH, -C(=0)NH ₂ , -C(=0)-NH-(d_ ₃ )alkyl, N0 ₂ , NH ₂ , -NH(d_ ₃ )alkyl), -N((d_ ₃ )alkyl) ₂ , -(d_ ₃ )alkyl-phenyl or phenyl optionally substituted with -(d_ ₃ )alkyl, -0-(d_ ₃ )alkyl or N0 ₂ .

R ³ -C: R ³ is H or (d_ ₃ )alkyl substituted with -C(=0)-N(R ³¹ )(R ³² );

R ³¹ is H or (d_ ₃ )alkyl;

R ³² is -(d_ ₃ )alkyl-phenyl, -(d_ ₃ )alkyl-Het, phenyl or Het;

wherein Het is defined as a 5- or 6-membered heteroaryl or heterocyclyl ring system;

wherein each said phenyl and Het, either alone or in combination with another radical, are optionally substituted 1 to 2 times with halo, -0-(d_ ₃ )alkyl, -(d_ ₃ )alkyl, -(d_ ₃ )haloalkyl, OH, COOH, -C(=0)-0-(d_ 3)alkyl, -C(=0)NH ₂ , NH ₂ , -NH(d_ ₃ )alkyl), -N((d_ ₃ )alkyl) ₂ , -S0 ₂ -(d_ 3)alkyl or -(d_ ₃ )alkyl-phenyl; and wherein said alkyl portion of alkyl-phenyl and alkyl-Het is optionally substituted 1 to 2 times with -C(=0)NH ₂ or -C(=0)-0-(Ci _-3 )alkyl; or R ³¹ and R ³² , together with the N to which they are attached, are linked to form a 5-, 6-, 9- or 10-membered Het optionally further containing 1 to 2 N heteroatoms;

wherein said Het is optionally substituted with 1 to 2 times with halo, - 0-(C _1-3 )alkyl, -(C _1-3 )alkyl optionally substituted with -C(=0)-0-(d_ 3)alkyl, -(d_ ₃ )haloalkyl, OH, COOH, -C(=0)NH ₂ , -C(=0)-NH-(d_ 3)alkyl, N0 ₂ , NH ₂ , -NH(d_ ₃ )alkyl), -N((d_ ₃ )alkyl) ₂ , -(d_ ₃ )alkyl-phenyl or phenyl optionally substituted with -(d_ ₃ )alkyl, -0-(d_ ₃ )alkyl or N0 ₂ .

The following table represents further embodiments E1 to E9 of the compounds of formula I, wherein each substituent group of each embodiment is defined according to the definitions set forth above:

In the aforementioned embodiments E1 -E9, if applicable, the following compounds are excluded:

4,5-dihydro-3,4-diphenyl-pyrrolo[3,4-c]pyrazol-6(1 H)-one;

4-(4-chlorophenyl)-4,5-dihydro-3-phenyl-pyrrolo[3,4-c]pyrazo l-6(1 H)-one;

4-(4-hydroxyphenyl)-4,5-dihydro-3-phenyl-pyrrolo[3,4-c]pyraz ol-6(1 H)-one;

4-(2-fluorophenyl)-4,5-dihydro-3-phenyl-pyrrolo[3,4-c]pyrazo l-6(1 H)-one;

4-(4-ethylphenyl)-4,5-dihydro-3-phenyl-pyrrolo[3,4-c]pyrazol -6(1 H)-one; and

4,5-dihydro-3-phenyl-4-(3-pyridinyl)-pyrrolo[3,4-c]pyrazo l-6(1 H)-one. Examples of most preferred compounds according to this invention are each single compound listed in Tables 1 to 4.

PHARMACEUTICAL COMPOSITION

Suitable preparations for administering the compounds of Formula (I) will be apparent to those with ordinary skill in the art and include for example tablets, pills, capsules, suppositories, lozenges, troches, solutions, syrups, elixirs, sachets, injectables, inhalatives and powders etc. The content of the pharmaceutically active compound(s) should be in the range from 0.05 to 90 wt.-%, preferably 0.1 to 50 wt.- % of the composition as a whole.

Suitable tablets may be obtained, for example, by mixing one or more compounds according to formula I with known excipients, for example inert diluents, carriers, disintegrants, adjuvants, surfactants, binders and/or lubricants . The tablets may also consist of several layers.

The dose range of the compounds of the invention applicable per day is usually from 0.01 to 100 mg/kg of body weight, preferably from 0.1 to 50 mg/kg of body weight. Each dosage unit may conveniently contain from 5% to 95% active compound (w/w). Preferably such preparations contain from 20% to 80% active compound.

The actual pharmaceutically effective amount or therapeutic dosage will of course depend on factors known by those skilled in the art such as age and weight of the patient, route of administration and severity of disease. In any case the combination will be administered at dosages and in a manner which allows a pharmaceutically effective amount to be delivered based upon patient's unique condition. COMBINATION THERAPY

When the composition of this invention comprises a combination of a compound of the invention and one or more additional therapeutic or prophylactic agent, both the compound and the additional agent should be present at dosage levels of between about 10 to 100%, and more preferably between about 10 and 80% of the dosage normally administered in a monotherapy regimen. Therefore, according to one embodiment, the pharmaceutical composition of this invention additionally comprises one or more antiviral agents.

Antiviral agents contemplated for use in such combination therapy include agents (compounds or biologicals) that are effective to inhibit the formation and/or replication of a virus in a human being, including but not limited to agents that interfere with either host or viral mechanisms necessary for the formation and/or replication of a virus in a human being. Such agents can be selected from:

• NRTIs (nucleoside or nucleotide reverse transcriptase inhibitors; including but not limited to zidovudine, didanosine, zalcitabine, stavudine, lamivudine, emtricitabine, abacavir, tenofovir, festinavir (OBP-601 ), elvucitabine, apricitabine);

• NNRTIs (non-nucleoside reverse transcriptase inhibitors; including but not limited to nevirapine, delavirdine, efavirenz, etravirine, rilpivirine, BILR 355, RDEA806, lersivirine (UK435061 ) and GSK2248761 (IDX-899));

· protease inhibitors (including but not limited to ritonavir, tipranavir, saquinavir, nelfinavir, indinavir, amprenavir, fosamprenavir, atazanavir, lopinavir, darunavir, brecanavir, TMC-31091 1 , PPL-100 (MK-8122), DG17 and SPI-256);

• entry inhibitors including but not limited to

• CCR5 antagonists (including but not limited to maraviroc (UK-427,857), vicriviroc (SCH-D, SCH-417690), TAK-652, INCB9471 , PF-232798, PRO-

140, TBR-652, SCH532706, GSK 706769 and TBR-220),

• CXCR4 antagonists (including but not limited to AMD-1 1070),

• fusion inhibitors (including but not limited to enfuvirtide (T-20), sifuvirtide, albuvirtide and TRI-1 144) and

· others (including but not limited to BMS-663069, ibalizumab (TNX-355) and BMS-488043);

• integrase inhibitors (including but not limited to raltegravir (MK-0518), c-1605, BMS-538158, elvitegravir (GS 9137), GSK1349572, GSK 1265744 and JTK- 656);

· TAT inhibitors;

• maturation inhibitors (including but not limited to bevirimat (PA-457)); and

• immunomodulating agents (including but not limited to levamisole).

Furthermore, a compound according to the invention can be used with at least one other compound according to the invention or with one or more antifungal or antibacterial agents (including but not limited to fluconazole).

EXAMPLES

Other features and advantages of the present invention will become apparent from the following more detailed Examples which illustrate, by way of example, the principles of the invention. As is well known to a person skilled in the art, reactions are performed in an inert atmosphere (including but not limited to nitrogen or argon) where necessary to protect reaction components from air or moisture. Solution percentages and ratios express a volume to volume relationship, unless stated otherwise. Flash chromatography is carried out on silica gel (Si0 ₂ ) according to the procedure of W.C. Still ef a/., J. Org. Chem., (1978), 43, 2923. Mass spectral analyses are recorded using electrospray mass spectrometry. Semi-preparative HPLC is carried out using a Combiprep ODS-AQ column, 50 x 20 mm, 5 μιη, 120 A, elution with a gradient of CH ₃ CN/H ₂ 0 containing 0.06% TFA.

Analytical HPLC is carried out under standard conditions with one of the following methods:

HPLC Method A: Combiscreen ODS-AQ C18 reverse phase column, YMC, 50 x 4.6 mm i.d., 5 μιη, 120 A at 220 nM, elution with a linear gradient of 2-100% CH ₃ CN / H ₂ 0 + 0.06 % TFA, run time = 10.5 min, flow = 3 mL/min. HPLC Method B: Waters Sunfire™ C18 3.5 μΜ reverse phase column, 4.6 x 50 mm i.d., 120 A at 220 nM, elution with a linear gradient of 5-100% CH ₃ CN / H ₂ 0 + 0.1 % TFA, run time = 10 min, flow = 3 mL/min.

HPLC Method C: Waters Sunfire™ C18 3.5 μΜ reverse phase column, 4.6 x 50 mm i.d., 120 A at 220 nM, elution with a linear gradient 2-100% CH ₃ CN /H ₂ 0 + 0.06 % TFA, run time = 10.5 min, flow = 3 mL/min.

Abbreviations or symbols used herein include:

Ac: acetyl; AcOH: acetic acid; BOC or Boc: fe/f-butyloxycarbonyl; Bu: butyl; CSA: capsid stabilization assay; DCM: dichloromethane; DDQ: 2,3-dichloro-5,6- dicyanobenzoquinone; DIPEA: diisopropylethylamine; DMAP: 4- dimethylaminopyridine; DME: ethylene glycol dimethyl ether; DMF: N,N- dimethylformamide; DMSO: dimethylsulfoxide; EC ₅ o: 50% effective concentration; Et: ethyl; Et ₃ N: triethylamine; EtOAc: ethyl acetate; EtOH: ethanol; Hex: hexane; HATU: N,N,N',N'-tetramethyl-0-(7-azabenzotriazol-1 -yl)uronium

hexafluorophosphate; HPLC: high performance liquid chromatography; IC ₅₀ : 50% inhibitory concentration; 'Pr or i-Pr: 1 -methylethyl (/so-propyl); LC-MS: liquid chromatography-mass spectrometry; m/z: mass-to-charge ratio; [M+H] ⁺ : protonated molecular ion; Me: methyl; MeOH: methanol; MS: mass spectrometry; MsCI:

methanesulfonyl chloride; Ph: phenyl; Pr: propyl; Prep LCMS: preparative liquid chromatography-mass spectrometry; RT: room temperature (approximately 18°C to 25°C); ferf-butyl, t-butyl or f-Bu: 1 , 1 -dimethylethyl; TFA: trifluoroacetic acid; Tf: trifluoromethanesulfonyl; THF: tetrahydrofuran; t _R : retention time. 1

1001

Step 1 :

(reference: Gein V.L., Kasimova N.N.; Russian Journal of General Chemistry, 2005, 75, 254-260)

A solution of 3-trifluoromethylbenzaldehyde (33 μΙ_, 0.25 mmol, Aldrich), ammonium acetate (87 mg, 1 .35 mmol, Aldrich) and ethyl 2,4-dioxo-4-phenylbutanoate (50 mg, 0.23 mmol, Aldrich) in EtOH (2 mL) is heated for 7 min at 120°C in a microwave apparatus (Biotage Initiator™). To the resulting solution is added AcOH (1 .5 mL) and hydrazine monohydrate (57 μΙ_, 1 .14 mmol, Aldrich). The reaction mixture is then heated for 1 1 min at 140°C in the microwave. The reaction mixture is diluted to 3 mL with AcOH, filtered and directly injected onto a semi-preparative HPLC to isolate product 1001.

Example 2: Preparation of compound 1007

1006 1007

Compound 1006 is prepared analogously to the procedure described in Example 1 . Step 1 :

Compound 1006 (13 mg, 0.04 mmol) is suspended in MeOH (1 mL) and thionylchloride (7 μΙ_, 0.10 mmol) is added at 0°C. The reaction mixture is heated to reflux for 18 h and concentrated under vacuum to dryness. The residue is dissolved in AcOH, filtered and directly injected onto a semi-preparative HPLC to isolate product 1007. 008

1006 1008

Compound 1006 is prepared analogously to the procedure described in Example 1 . Step 1 :

Compound 1006 (15 mg, 0.047 mmol) is dissolved in DMF (0.5 mL) and methyl amine (2 M) in THF (52 μΙ_, 0.103 mmol), diisopropylethylamine (16 μΙ_, 0.94 mmol) and HATU (23 mg, 0.061 mmol) are sequentially added. The reaction mixture is stirred for 3 h at RT, acidified with a 6 N HCI solution and directly injected onto a semi-preparative HPLC to isolate product 1008.

Example 4: Preparation of compound 1028

1028

Compound 1026 is prepared analogously to the procedure described in Example 1. Step 1 :

Compound 1026 (707 mg, 2.21 mmol) is dissolved in THF (1 1 mL) and di-ferf-butyl dicarbonate (506 mg, 2.32 mmol) followed by 4-(dimethylamino)pyridine (270 mg, 2.21 mmol) are added. The reaction mixture is stirred at RT for 1 h and diluted with EtOAc. The organic phase is washed with a saturated sodium bicarbonate solution and with brine. The organic solution is dried with MgS0 ₄ , filtered and concentrated under vacuum to dryness. The crude material is purified by flash chromatography (40% EtOAc in Hex) to isolate product 4.1.

Step 2:

Compound 4.1 (531 mg, 1.26 mmol) is suspended in MeOH (32 mL) and then palladium hydroxide (80 mg) is added. The reaction mixture is stirred under a hydrogen atmosphere for 40 min. The reaction mixture is filtered through Celite™ and concentrated under vacuum to dryness to isolate product 4.2.

Step 3:

Compound 4.2 (30 mg, 0.077 mmol) and benzenesulfonyl chloride (1 1 μί, 0.085 mmol, Aldrich) are dissolved in pyridine (0.5 mL) and the reaction mixture is stirred at RT for 1 h. The reaction mixture is then concentrated under vacuum to dryness and 4 N HCI in dioxane (2 mL) is added to the residue. The mixture is stirred for 16 h at RT and is then concentrated under vacuum to dryness. The residue is dissolved in AcOH, filtered and directly injected onto a semi-preparative HPLC to isolate product 1028.

Step 1 :

Compound 4.2 (30 mg, 0.077 mmol) is dissolved in DCM (0.5 mL) and treated with phenyl isocyanate (9 μΙ_, 0.085 mmol, Aldrich). The reaction mixture is stirred at RT for 30 min. The reaction mixture is then concentrated under vacuum to dryness and 4 N HCI in dioxane (2 mL) is added to the residue. The mixture is stirred for 16 h at RT and is concentrated under vacuum to dryness. The residue is dissolved in AcOH, filtered and directly injected onto a semi-preparative HPLC to isolate product 1029.

Example 6: Preparation of compound 1033

Step 1 :

Potassium carbonate (679 mg, 4.91 mmol) is added to a solution of 4- hydroxybenzaldehyde (400 mg, 3.28 mmol, Aldrich) and 4-nitrobenzyl bromide (708 mg, 3.28 mmol, Aldrich) in DMF (4 mL). The reaction mixture is then stirred for 15 min at 100°C and cooled to RT. The reaction mixture is diluted with EtOAc and washed with water. The organic phase is dried over sodium sulfate, filtered and concentrated under vacuum to dryness. The crude material is purified by flash chromatography (100% Hex to 50% EtOAc in Hex) to isolate product 6.1. Step 2:

(reference: Gein V.L., Kasimova N.N.; Russian Journal of General Chemistry, 2005, 75, 254-260)

A solution of compound 6.1 (705 mg, 2.74 mmol), ammonium acetate (1.06 g, 13.7 mmol, Aldrich) and methyl 2,4-dioxo-4-phenylbutanoate (604 mg, 2.93 mmol, Bionet) in EtOH (9 mL) is heated for 7 min at 120°C in a microwave apparatus (Biotage Initiator™). To the resulting solution is added AcOH (9 mL) and hydrazine monohydrate (686 μί, 13.7 mmol, Aldrich). The reaction mixture is then heated for 1 1 min at 140°C in the microwave. The reaction mixture is cooled to RT and the resulting solid is collected by filtration. The solid is washed with EtOH and dried under vacuum to afford compound 1033.

Example 7: Preparation of compound 1037

1033 1037

Step 1 :

To compound 1033 (35 mg, 0.082 mmol) in EtOH (1 mL) and water (0.25 mL) are added iron (18 mg, 0.33 mmol) and 1 N HCI (82 μί, 0.082 mmol). The reaction mixture is heated for 2 h at 100°C in a sealed vial. The reaction mixture is filtered and directly injected onto a semi-preparative HPLC to isolate product 1037.

Example 8: Preparation of compound 1040

1033 1040 Step 1 :

To a suspension of compound 1033 (35 mg, 0.082 mmol) in EtOH (0.5 mL), acetic anhydride (1 mL) and water (0.25 mL) is added iron (18 mg, 0.33 mmol) and 1 N HCI (82 μί, 0.082 mmol). The reaction mixture is heated for 30 min at 100°C in a sealed vial. The reaction mixture is filtered and directly injected onto a semi- preparative HPLC to isolate product 1040. 1034

9.1 1034

Compound 9.1 is prepared analogously to the procedure described in Example 1

Step 1 :

Compound 9.1 (35 mg, 0.082 mmol) is suspended in MeOH (2 mL) and palladium hydroxide (6 mg) is added. The reaction mixture is stirred under a hydrogen atmosphere for 45 min. The reaction mixture is filtered through Celite™ and concentrated under vacuum to dryness. The residue is dissolved in AcOH, filtered and directly injected onto a semi-preparative HPLC to isolate product 1034.

1026 10.1 1041

Compound 1026 is prepared analogously to the procedure described in Example 1. Step 1 :

Compound 1026 (200 mg, 0.62 mmol) is suspended in MeOH (2 mL) and palladium hydroxide (40 mg) is added. The reaction mixture is stirred under a hydrogen atmosphere for 45 min. The reaction mixture is filtered through Celite and concentrated under vacuum to dryness to isolate compound 10.1.

Step 2:

To compound 10.1 (30 mg, 0.10 mmol) and benzaldehyde (23 μΙ_, 0.22 mmol) in / ^' - P/OAc is added sodium borohydride (55 mg, 0.25 mmol) and AcOH (12 μΙ_). The reaction mixture is stirred for 24 h at 60°C and then diluted with EtOAc. The organic solution is washed with 1 N NaOH and brine. The organic phase is dried over MgS0 ₄ , filtered and concentrated under vacuum to dryness. The residue is dissolved in AcOH, filtered and directly injected onto a semi-preparative HPLC to isolate product 1041.

Example 11 : Preparation of compound 1045

Step 1 :

3-Nitropropiophenone (300 mg, 1.67 mmol, Aldrich) is dissolved in MeOH (16 mL) and cooled to 0°C. Sodium borohydride (76 mg, 2.00 mmol) is added and the reaction mixture is stirred for 1 h at RT. The reaction mixture is concentrated under vacuum to dryness. The crude material is purified by flash chromatography (25% EtOAc in Hex to 35% EtOAc in Hex) to isolate product 11.1.

Step 2:

To a solution of compound 11.1 (52 mg, 0.29 mmol) in DCM (1.5 mL) is added Et ₃ N (40 μΙ_, 0.29 mmol) and MsCI (22 μΙ_, 0.29 mmol) and the reaction mixture is stirred for 2 h at 0°C. The reaction mixture is diluted with EtOAc and washed with a saturated sodium bicarbonate solution. The organic phase is dried over sodium sulfate, filtered and concentrated under vacuum to dryness. The crude material is purified by flash chromatography (20% EtOAc in Hex to 40% EtOAc in Hex) to isolate product 11.2.

Steps 3 and 4:

Performed analogously to the procedure described in Example 6. Step 5:

Performed analogously to the procedure described in Example 7 to afford compound 1045.

1044

Step 1 :

4-Aminobenzaldehyde (125 mg, 1.03 mmol, Betapharma), isovaleryl chloride (135 μΙ_, 1.11 mmol, Aldrich) and DIPEA (550 μΙ_, 3.16 mmol) are dissolved in DCM (10 mL) and stirred for 1 h at RT. Water is added and the phases are separated. The organic phase is washed with an aqueous citric acid solution and with brine, dried over sodium sulfate, filtered and concentrated under vacuum to dryness to isolate product 12.1. Product 12.1 is used as such without any purification.

Step 2:

Performed analogously to the procedure described in Example 1 to afford compound 1044.

Example 13: Preparation of compound 1047

Step 1 :

(3-Aminocarbonylphenyl)boronic acid (64 mg, 0.390 mmol, Frontier), 3-bromo-4- hydroxybenzaldehyde (53 mg, 0.260 mmol), potassium carbonate (90 mg, 0.650 mmol) and PEPPSI ^(TM) -/Pr catalyst (18 mg, 0.026 mmol, Aldrich) are suspended in DMF (0.7 mL) under argon. The reaction mixture is heated to 80°C for 16 h, filtered and directly injected onto a semi-preparative HPLC to isolate product 13.1.

Step 2:

A solution of compound 13.1 (16 mg, 0.067 mmol), ammonium acetate (21 mg, 0.27 mmol, Aldrich) and methyl 2,4-dioxo-4-phenylbutanoate (1 1 mg, 0.053 mmol, Aldrich) in EtOH (1.5 mL) is heated for 7 min at 120°C in a microwave apparatus (Biotage Initiator™). The resulting mixture is concentrated under vacuum to dryness. AcOH (1.5 mL) and hydrazine monohydrate (1 1 μί, 0.27 mmol, Aldrich) are added and the reaction mixture is heated for 1 1 min at 140°C in the microwave. The reaction mixture is filtered and directly injected onto a semi-preparative HPLC to isolate product 1047.

Example 14: Preparation of compounds 1050, 1051 and 1053

Step 1 :

To 3-nitrophenacylamine hydrochloride (1.0 g, 4.6 mmol, Alfa Aesar) in THF (23 mL) is added DIPEA (0.88 mL, 5.1 mmol), di-ferf-butyl dicarbonate (1.0 g, 4.6 mmol, Oakwood) and DMAP (0.56 g, 4.6 mmol). The reaction mixture is stirred at RT for 1 h and diluted with EtOAc. The reaction mixture is washed with water, 1 N HCI, saturated sodium bicarbonate and brine. The organic phase is dried over MgS0 ₄ , filtered and concentrated under vacuum to dryness to isolate product 14.1. Product 14.1 is used as such without any purification.

Steps 2 to 4:

Performed analogously to the procedure described in Example 1 1 (steps 1 to 3). Step 5:

(reference: Gein V.L., Kasimova N.N.; Russian Journal of General Chemistry, 2005, 75, 254-260)

A solution of compound 14.4 (35 mg, 0.091 mmol), ammonium acetate (35 mg, 0.46 mmol, Aldrich) and methyl 2,4-dioxo-4-phenylbutanoate (20 mg, 0.091 mmol, Aldrich) in EtOH (1 mL) is heated for 7 min at 120°C in a microwave apparatus (Biotage Initiator™). To the resulting solution is added AcOH (1 mL) and hydrazine monohydrate (22 μΙ_, 0.46 mmol, Aldrich). The reaction mixture is then heated for 1 1 min at 140°C in the microwave and concentrated under vacuum to dryness. To the residue in DCM (1 mL) is added TFA (140 μΙ_, 1.8 mmol) and the reaction mixture is stirred for 16 h at RT. The reaction mixture is concentrated under vacuum to dryness and the residue is dissolved in AcOH, filtered and directly injected onto a semi-preparative HPLC to isolate product 14.5.

Step 6:

To compound 14.5 (1 1 mg, 0.023 mmol) in DMF (1 mL) is added AcOH (1.3 μί, 0.023 mmol), DIPEA (5.2 μΙ_, 0.030 mmol) and HATU (1 1 mg, 0.028 mmol). The reaction mixture is stirred at RT for 30 min. It is then diluted with EtOAc and washed with 1 N HCI, saturated sodium bicarbonate and brine. The organic solution is dried over MgS0 ₄ , filtered and concentrated under vacuum to dryness to isolate product 14.6. Product 14.6 is used as such without any purification.

Step 7:

To compound 14.5 (16 mg, 0.035 mmol) in pyridine (0.5 mL) is added methyl chloroformate (2.7 μί, 0.035 mmol). The reaction mixture is stirred at RT for 4 h. The reaction mixture is concentrated under vacuum to dryness and the residue is dissolved in AcOH, filtered and directly injected onto a semi-preparative HPLC to isolate product 14.7.

Steps 8, 9 and 10:

Performed analogously to the procedure described in Example 7. 54

1036 1054

Compound 1036 is prepared analogously to the procedure described in Example 7. Step 1 :

To compound 1036 (18 mg, 0.047 mmol) in TFA (0.25 mL) is added sodium nitrite (3.2 mg, 0.047 mmol) at 0°C. The reaction mixture is stirred for 15 min and sodium azide (9.1 mg, 0.14 mmol) is then added (N ₂ evolution). The mixture is stirred at 0°C for 1 h, diluted with water and then filtered. The crude material is dissolved in AcOH, filtered and directly injected onto a semi-preparative HPLC to isolate product 1054. 070

16.1 1070

Compound 16.1 is prepared analogously to the procedure described in Example 13.

Step 1 :

To compound 16.1 (44 mg, 0.095 mmol) in DCM (1 mL) at 0°C is added 1 M BBr ₃ in DCM (0.14 mL, 140 mmol). The reaction mixture is stirred for 30 min at 0°C and diluted with EtOAc. It is then washed with water and saturated sodium bicarbonate. The organic solution is dried over MgS0 ₄ , filtered and concentrated under vacuum to dryness. The crude material is dissolved in AcOH, filtered and directly injected onto a semi-preparative HPLC to isolate product 1070. 1072

1034 1072

Step 1 :

To compound 1034 (16 mg, 0.052 mmol) in MeOH (0.26 mL) is added

benzaldehyde (5.3 μί, 0.052 mmol) and the reaction mixture is stirred for 6 h at 45°C. The reaction mixture is concentrated under vacuum to dryness and the crude material is dissolved in DCM (0.52 mL). DDQ (13 mg, 0.057 mmol) is then added and the reaction mixture is stirred at RT for 45 min. The reaction mixture is diluted with DCM and washed twice with 10% sodium carbonate and brine. The organic solution is dried over MgS0 ₄ , filtered and concentrated under vacuum to dryness. The crude material is dissolved in AcOH, filtered and directly injected onto a semi- preparative HPLC to isolate product 1072.

1075 18.2 1076

Step 1 :

To 3-nitrobenzyl alcohol (108 μΙ_, 0.71 mmol, Aldrich) in DMF (1 mL) is added NaH 60% in mineral oil (28 mg, 0.71 mmol) and the reaction mixture is stirred for 5 min at RT. 6-chloropyridine-3-carboxaldehyde (100 mg, 0.71 mmol, Aldrich) is added and the reaction mixture is stirred for 30 min at 1 10°C. The reaction mixture is diluted with EtOAc and washed with a saturated sodium bicarbonate solution. The organic phase is dried over sodium sulfate, filtered and concentrated under vacuum to dryness. The crude material is purified by flash chromatography (100% Hex to 100% EtOAc) to isolate product 18.1.

Step 2:

Performed analogously to the procedure described in Example 1 to afford 18.2.

Step 3: Performed analogously to the procedure described in Example 7 to afford compound 1075.

Step 4:

Performed analogously to the procedure described in Example 8 to afford compound 1076.

2005

Step 1 :

To 3'-hydroxyacetophenone (1.0 g, 7.4 mmol, Acros) and 4-methylbenzyl bromide (1.4 g, 7.6 mmol, Aldrich) in DMF (8 mL) is added cesium carbonate (2.5 g, 7.7 mmol) and the reaction mixture is heated at 100°C for 18 h. The mixture is poured into 1 M HCI and extracted with EtOAc. The organic phase is washed with 1 M HCI and brine, dried with MgS0 ₄ , filtered and concentrated under vacuum to dryness. The crude material is purified by flash chromatography (5% EtOAc in Hex to 10% EtOAc in Hex) to isolate product 19.1.

Step 2:

Compound 19.1 (1.0 g, 4.2 mmol) and diethyl oxalate (1.1 mL, 8.1 mmol, Aldrich) in THF (6 mL) are added dropwise to a solution of sodium fe/f-butoxide (1.6 g, 16 mmol) in THF (14 mL) at RT. The reaction mixture is then poured over a 0.5 N HCI solution and the aqueous solution is extracted with EtOAc. The organic phase is dried with MgS0 ₄ , filtered and concentrated under vacuum to dryness. The crude material is purified by flash chromatography (25% EtOAc in Hex) to isolate product 19.2. Step 3:

Performed analogously to the procedure described in Example 1 to afford compound 2005.

2017

Step 1 :

To 2-thiophenemethanol (420 mg, 3.1 mmol, Aldrich), 3'-hydroxyacetophenone (0.37 mL, 3.9 mmol, Acros) and triphenylphosphine (1.2 g, 4.5 mmol) in THF (10 mL) at 0°C is added diisopropyl azodicarboxylate (0.91 mL, 4.5 mmol, Aldrich) in THF (5 mL) dropwise over 30 min. The reaction mixture is stirred for 3 h at 0°C and then for 24 h at RT. The reaction mixture is concentrated under vacuum to dryness and the crude material is purified by flash chromatography (100% Hex to 30% EtOAc in Hex) to isolate product 20.1.

Steps 2 and 3:

Performed analogously to the procedure described in Example 19 (Steps 2 and 3). Example 21 : Preparation of compound 2023

2023 21.3

Step 1 :

To 3'-iodoacetophenone (1.9 g, 7.6 mmol, Transworld), phenylacetylene (0.88 mL, 7.8 mmol, Aldrich), diethylamine (1.8 mL, 17 mmol, Aldrich) and copper(l)iodide (145 mg, 0.76 mmol, Aldrich) in THF (20 mL) is added Pd(PPh ₃ ) ₄ (860 mg, 0.74 mmol, Strem) and the reaction mixture is stirred for 16 h at 75°C. The reaction mixture is diluted with EtOAc and washed with a saturated sodium bicarbonate solution, water and brine. The organic phase is dried with MgS0 ₄ , filtered and concentrated under vacuum to dryness. The crude material is purified by flash chromatography (10% EtOAc in Hex) to isolate product 21.1.

Step 2:

To a solution of compound 21.1 (678 mg, 3.08 mmol) in EtOH (15 mL) is added palladium hydroxide 20% w/w (50 mg, Aldrich) and the reaction mixture is stirred for 5 h at RT under an atmosphere of hydrogen. The catalyst is removed by filtration through a Millex™ filter and the solution is concentrated under vacuum to dryness. The crude material is purified by flash chromatography (5% EtOAc in Hex) to isolate product 21.2.

Steps 3 and 4:

Performed analogously to the procedure described in Example 19 (Steps 2 and 3). Example 22: Preparation of compound 2030

2030 22.3

Step 1 :

Performed analogously to the procedure described in Example 20, Step 1

Step 2:

Compound 22.1 (150 mg, 0.49 mmol), 2,4,6-trivinylcyclotriboroxane pyridine complex (120 mg, 0.50 mmol, Aldrich) and Pd(PPh ₃ ) ₄ (60 mg, 0.05 mmol, Strem) in DME (5 mL) and 2 M sodium bicarbonate (5 mL) are purged with argon (3x). The reaction mixture is heated at 80°C under an argon atmosphere for 16 h. The reaction mixture is diluted with EtOAc and washed with water. The organic phase is dried over sodium sulfate, filtered and concentrated under vacuum to dryness. The crude material is purified by flash chromatography (5% EtOAc in Hex to 30% EtOAc in Hex) to isolate product 22.2.

Steps 3 and 4:

Performed analogously to the procedure described in Example 19 (Steps 2 and 3). Example 23: Preparation of compounds 1055 and 3001

Step 1 :

(reference: Gein V.L., Kasimova N.N.; Russian Journal of General Chemistry, 2005, 75, 254-260)

To a solution of 3-pyridinecarboxaldehyde (2.36 g, 25 mmol, Aldrich) in EtOH (230 mL) is added ammonium acetate (8.75 g, 1 14 mmol). The reaction mixture is heated to reflux for 1 h. Ethyl 2,4-dioxo-4-phenylbutanoate (5.0 g, 23 mmol, Aldrich) is added and the reflux is continued for 1 h. Hydrazine monohydrate (5.5 mL, 1 13 mmol) is added and the reflux is continued for 2 h. AcOH (230 mL) and TFA (4.7 mL) are added and reflux is continued for 5 h. The reaction mixture is concentrated under vacuum to dryness and a saturated sodium carbonate solution is added until basic pH is achieved. The reaction mixture is extracted with EtOAc (4x) and the combined organic phases are washed with brine. The organic solution is dried over MgS0 ₄ , filtered and concentrated under vacuum to dryness. The crude material is purified by flash chromatography (5% EtOH in EtOAc to 10% EtOH in EtOAc) to isolate product 1055.

Step 2:

To a solution of compound 1055 (4.0 g, 15 mmol) in DMF (15 mL) at 0°C is added NaH 60% in mineral oil (720 mg, 18 mmol). The reaction mixture is stirred for 20 min at 0°C and methyl bromoacetate (1.6 mL, 17 mmol) is added. The reaction mixture is stirred for 3 h at RT and poured into iced water. The pH is adjusted to 4-5 with 1 N HCI and the aqueous mixture is extracted with EtOAc. The combined organic phases are washed with brine, dried over MgS0 ₄ , filtered and concentrated under vacuum to dryness to isolate compound 23.1. Compound 23.1 is used as such without purification.

Step 3:

To a solution of compound 23.1 (4.7 g, 14 mmol) in MeOH (20 mL) is added 5 M NaOH (18 mL, 90 mmol) and the reaction mixture is stirred for 2 h at RT. The pH of the solution is adjusted to 4-5 with 1 N HCI and the reaction mixture is diluted with EtOAc. The organic phase is washed with brine, dried over MgS0 ₄ , filtered and concentrated under vacuum to dryness to isolate compound 23.2. Compound 23.2 is used as such without purification.

Step 4:

To a solution of compound 23.2 (8.4 mg, 0.025 mmol) in DMF (0.1 mL) are successively added 3-aminopyridine (2.8 mg, 0.030 mmol), HATU (1 1 mg, 0.30 mmol) and Et ₃ N (17 μί, 0.12 mmol). The reaction mixture is stirred for 2 h at RT. The reaction mixture is filtered and directly injected onto a semi-preparative HPLC to isolate product 3001.

Step 1 :

To a solution of compound 1055 (500 mg, 1.8 mmol) in DMF (3 mL) at 0°C is added NaH 60% in mineral oil (87 mg, 2.2 mmol). The reaction mixture is stirred for 20 min at 0°C, allyl bromide (0.19 mL, 2.2 mmol) is then added and stirring is continued for 3 h at RT. The reaction mixture is acidified with a potassium hydrogen sulfate solution and diluted with EtOAc. The phases are separated and aqueous phase is extracted with EtOAc. The combined organic phases are washed with brine, dried over MgS0 ₄ , filtered and concentrated under vacuum to dryness to isolate compound 24.1. Compound 24.1 is used as such without purification.

Step 2:

To a solution of compound 24.1 (100 mg, 0.32 mmol) in THF (3.3 mL) is added water (2.8 mL), osmium tetroxide 2.5% in fe/f-butanol (46 μΙ_, 0.005 mmol) and sodium periodate (220 mg, 0.95 mmol). The reaction mixture is stirred for 16 h at RT and concentrated under vacuum to dryness. EtOAc is added to the residue and the organic mixture is washed with water and brine. The organic phase is dried over MgS0 ₄ , filtered and concentrated under vacuum to dryness to isolate compound 24.2. Compound 24.2 is used as such without purification.

Step 3:

To a solution of compound 24.2 (25 mg, 0.079 mmol) in DMF (0.5 mL) is added dimethylamine hydrochloride (24 mg, 0.30 mmol, Aldrich) and the reaction mixture is stirred for 1 h at RT. Sodium cyanoborohydride (12 mg, 0.19 mmol, Aldrich), AcOH (9 μί, 0.15 mmol) and sodium acetate (25 mg, 0.30 mmol) are added and stirring is continued for 2 days. The reaction mixture is filtered and directly injected onto a semi-preparative HPLC to isolate product 3002.

4001

Step 1 :

To benzaldehyde (0.25 mL, 2.5 mmol, Aldrich) in EtOH (20 mL) is added ammonium acetate (875 mg, 1 1 mmol) and the reaction mixture is heated to reflux for 1 h. Ethyl 2,4-dioxo-4-phenylbutanoate (500 mg, 2.3 mmol, Aldrich) is added and the reflux is continued for 4 h. The reaction mixture is cooled to RT and the resulting solid is collected by filtration. The solid is washed with MeOH to afford compound 25.1.

Step 2:

(reference: Gein V.L., Kasimova N.N.; Russian Journal of General Chemistry, 2005, 75, 254-260)

To compound 25.1 (50 mg, 0.18 mmol) in AcOH (2 mL) is added 2- hydroxyethylhydrazine (61 μί, 0.90 mmol, Aldrich) and the reaction mixture is heated for 1 1 min at 140°C in a microwave apparatus (Biotage Initiator™). The reaction mixture is filtered and directly injected onto a semi-preparative HPLC to isolate product 4001.

Compound 26.1 is prepared analogously to the procedure described in Example 20 (Steps 1 and 2).

Step 1 :

Performed analogously to the procedure described in Example 1. Steps 2, 3 and 4:

Performed analogously to the procedure described in Example 23 (Steps 2, 3 and 4).

Step 5:

To a solution of compound 26.5 (28 mg, 0.041 mmol) in EtOH (1 mL) is added iron (9 mg, 0.16 mmol) and 1 N HCI (41 μΙ_, 0.41 mmol). The reaction mixture is stirred at 80°C for 45 min and diluted with AcOH. The reaction mixture is filtered and directly injected onto a semi-preparative HPLC to isolate product 4003.

2003 2007 2008

Compound 2003 is prepared analogously to the procedure described in Example 19.

Step 1 :

The two enantiomers of compound 2003 are separated using chiral semi- preparative HPLC (Chiracel-AGP (150x10 mm) 5 micron, isocratic 12% ACN / 88% H ₂ 0 + 0.1 % AcOH (pH = 4.3), 4 mL/min, 60 min) to isolate compounds 2007 and 2008.

Example 28: Capsid Stabilization Assay

The GAG polyprotein is the major structural protein required for HIV viral particle assembly. During the maturation process, Gag is cleaved by the viral protease and releases its four major proteins, matrix (MA), capsid (CA), nucleocapsid (NC) and p6, as well as two spacer peptides termed SP1 and SP2. This triggers profound morphological changes to the viral particle as CA will reassemble to form the viral core, a structure that is essential for viral infectivity. The compounds of the invention inhibit HIV-1 capsid disassembly as tested using an immobilized capsid stabilization assay (CSA) using the CA-NC polypeptide of SEQ ID NO: 2 having a G94D mutation over the wildtype (numbered based on SEQ ID NO: 2) encoded by SEQ ID NO: 1 .

SEQ ID NO: 1 is a nucleic acid of 900 base pairs encoding HIV-1 NL4-3 CA-NC (Gag residues 133-432), having the CA G94D mutation of SEQ ID NO: 2. SEQ ID NO: 1 , is transferred to pET-1 1 a expression vector (Novagen™) by PCR amplification using primers that introduce an Ndel site and a start codon at the 5'-end and a BamHI site and a stop codon at the 3'-end. Resistance mutations in CA were identified by passage of the HIV-1 virus in the presence of compound 1027, namely, A105T, T107A and T107N, all numbered with reference to SEQ ID NO: 2,. These resistance mutations may be introduced into the expression vector using the QuikChange® II Site-Directed Mutagenesis Kit (Stratagene) according to the manufacturer's instructions.

SEQ ID NO: 2 is expressed in the BL21 (DE3) E. coli cells (Novagen™). Briefly, LB media is inoculated with overnight pre-cultures and grown at 37°C until mid log- phase (Abs600 ~0.6), protein expression is induced by addition of 0.5-1 mM isopropyl-p,D-thiogalactopyranoside (IPTG) and carried out for 4 to 6 hours at 30°C. Cells are harvested by centrifugation and pellets are stored at -80°C until purification.

Purification SEQ ID NO: 2 is as follows: 5 to 10 g of cell paste are lysed by sonication in 40 mL of Buffer A [20 mM Tris pH 7.5; 1 μΜ ZnCI2; 10 mM β- mercaptoethanol] supplemented with 0.5M NaCI and Complete EDTA-free® protease inhibitors tablets (Roche). Nucleic acids and cell debris are removed by adding 0.1 1 volumes of 0.2 M ammonium sulfate and an equivalent volume of 10% poly-ethyleneimine pH 8.0, stirring the sample for 20 minutes at 4°C, followed by centrifugation at 30 OOOx g for 20 minutes. SEQ ID NO: 2 is recovered from the supernatant by adding 0.35 volumes of saturated ammonium sulfate solution followed by centrifugation at 10 OOOx g for 15 minutes. The pellet is dissolved in 10 ml of Buffer A + 0.1 M NaCI and dialyzed overnight in the same buffer but at 0.05 M NaCI (Dialysis Buffer). The sample is then cleared by centrifugation and passed on a 1-ml HiTrap™ SP HP column (GE Healthcare) pre-equilibrated with Dialysis Buffer. SEQ ID NO: 2 is eluted with Buffer A supplemented with 0.5 M NaCI and fractions containing the protein are pooled. The absorbance of SEQ ID NO: 2 at 280nm is measured and the concentration is determined using the calculated molar extinction coefficient (E = 40220 M ^"1 cm ^"1 ).

Capsid Stabilization Assay (CSA)

Reacti-Bind® Neutravidin Coated black 384-well plates (Pierce, catalogue # 15402) are washed once with 80 μΙ/well of Buffer B (50 mM Tris pH 8.0; 350 mM NaCI; 10 μΜ ZnS04; 0.0025% CHAPS (w/v); 50 μg/ml BSA; 1 mM DTT). Immobilization of a 5'-end biotin labeled (TG)25 oligonucleotide (Integrated DNA Technology Inc.) is carried out by adding 50 μΙ/well of a 25 nM solution of oligonucleotide in Buffer C (50 mM Tris pH 8.0; 350 mM NaCI; 10 μΜ ZnS04; 0.0025% CHAPS (w/v); 5mg/ml BSA; 1 mM DTT) and incubating overnight. Unbound material is removed by washing twice with 80 μΙ/well of Buffer B. Assembly reactions are performed in 60 μΙ/well of Buffer B comprising 100 nM of 5'-end fluorescein labeled (TG)25 oligonucleotide (Integrated DNA Technology Inc.) and 2 μΜ SEQ ID NO: 2 Assembly reactions are incubated for 2 hours at RT and non-immobilized material is removed by washing once with 80 μΙ/well of Buffer D (50 mM Tris pH 8.0; 250 mM NaCI; 10 μΜ ZnS04; 0.0025% CHAPS (w/v); 50 μ9/ητιΙ BSA; 1 mM DTT). The negative control wells are washed once with 80 μΙ/well of Buffer B and 80 μΙ/well of Buffer B supplemented with 0.1 % SDS (w/v) is added. The wells are incubated for 15 minutes prior to the quantification of captured fluorescence on a Victor2 plate reader (Perkin Elmer Life Sciences) equipped with fluorescein excitation and emission filters using manufacturer's setting for fluorescein fluorescence. The remainder of the microplate is processed as follows: Buffer D is removed and test compounds, serially diluted in Buffer D + 1 % dimethyl sulfoxide (DMSO), are added to the wells (60 μΙ/well) and the microplate is incubated at room temperature for 2 hours. The positive control wells lack compounds (1 % DMSO only) and are treated identically to wells receiving test compounds. Disassembled material is removed by two successive 80 μΙ/well washes with Buffer B. Finally, 80 μΙ/well of Buffer B + 0.1 % SDS (w/v) is added and the microplate is incubated for 15 minutes prior to quantification of captured fluorescence as described above. The capacity of a test compound to inhibit the dissociation of the assembled complexes is considered proportional to the observed gain of captured fluorescence. For each well, the % disassembly is calculated using the following equation:

% disassembly = (F _ne g control ^— Ftest well) (Fneg control ^— Fpositive control) ^* 100, where F _neg control is the average fluorescence of all the negative control wells within the assay plate, F _p0S itive control is the average fluorescence of all the positive control wells within the assay plate and F _te st weii is average fluorescence of the wells with test compound. For the negative and positive control wells, the % disassembly is 0 and 100%, respectively.

The % disassembly values are then used to generate an IC ₅ o value by fitting the values from the ten-point dilution series to the following equation:

% inhibition = ((Imaxn χ [l]n) ÷ ([l]n + IC50n)) where the IC ₅ o represents the concentration of compound required to inhibit the dissociation of 50% of the assembled complexes. IC ₅₀ data for representative compounds is provided in the table below:

Example 29: C8166 HIV-1 Luciferase Assay (EC ₅₀ )

The assay used to measure inhibition of HIV replication is as described in WO 2004/050643, pages 73-75, herein incorporated by reference, with the following modifications:

Preparation of test compounds

Serial dilutions of test compounds are prepared in RPMI 1640 media supplemented with 10% fetal bovine serum and 1 % penicillin/streptomycin (referred to here-in as complete media) from DMSO stock solutions. Eleven serial dilutions of the test compounds are prepared and the 12 ^th well contains complete media with no test compound and serves as the positive control. All samples, including the negative and positive controls, contain the same concentration of DMSO (< 0.5%) in complete media. 100 μΙ of diluted test compound is added, to triplicate wells, of a 96 well assay plate (Corning Costar black microtiter plate, catalogue # 3904).

Infection of cells

The C8166-LTR-L.UC cells are infected with HIV-1 at a moi of 0.005 in a minimal volume of complete media in a tissue culture flask (ex. 3x107 cells in 10 mL of complete media/25 cm2 flask) for 1 .5 hours at 37°C in a 5% C02 incubator (with rocking). The infected cells are then diluted with complete media, to a final concentration of 2.5x105 cells/ml. 100 μΙ of diluted cells are then added to each well of the assay plate (2.5x104 cells/well) containing the test compounds. 2.5x104 uninfected cells/well are added to the last row of the assay plate, in a final volume of 200 μΙ and serve as the negative control. Cells are incubated for 3 days at 37°C in an incubator containing 5% C02. Firefly luciferase activity is determined by adding 50 μΙ of SteadyGlo (ProMega, catalogue #E2520 ) to each well of the assay plate. Luminescence is then measured using the LumiStar Galaxy plate reader (BMG Labtech). % inhibition values are used to generate an EC50 value which represents the concentration of test compound that inhibits 50% of HIV-1 viral replication. EC50 data for representative compounds is provided in the table of Example 28.

TABLES OF COMPOUNDS

The following tables list compounds representative of the invention. All of the compounds in Tables 1 to 4 are synthesized analogously to the Examples described above. For each compound in the tables, the analogous synthetic route to prepare each compound is identified by Example number. It will be apparent to a skilled person that the analogous synthetic routes may be used, with appropriate modifications, to prepare the compounds of the invention as described herein.

Retention times (t _R ) for each compound are measured using the standard analytical HPLC conditions described in the Examples. As is well known to one skilled in the art, retention time values are sensitive to the specific measurement conditions. Therefore, even if identical conditions of solvent, flow rate, linear gradient, and the like are used, the retention time values may vary when measured, for example, on different HPLC instruments. Even when measured on the same instrument, the values may vary when measured, for example, using different individual HPLC columns, or, when measured on the same instrument and the same individual column, the values may vary, for example, between individual measurements taken on different occasions.

The compounds listed in Tables 1 to 4 showed either EC ₅₀ values in the range of 40 μΜ or less, and mostly in a range of 15 μΜ or less, when tested in the assay of Example 29 or showed IC ₅₀ values in the range of 40 μΜ or less, and mostly in a range of 20 μΜ or less, when tested in the assay of Example 28.

TABLE 1

TABLE 2

HPLC Synthetic

Cmpd # R ¹ _R 2a tp (M+H) ⁺

(min) method method

HO

2029 5.3 B 412.2 Ex. 20

2030 4.981 A 409.2 Ex. 22

Ex. 15 &

2031 5.58 B 467.1

19

TABLE 3

TABLE 4

Each reference, including all patents, patent applications, and publications cited in the present application is incorporated herein by reference in its entirety, as if each of them is individually incorporated. Further, it would be appreciated that, in the above teaching of invention, the skilled in the art could make certain changes or modifications to the invention, and these equivalents would still be within the scope of the invention defined by the appended claims of the application.