KR20130127515A - Combinations of a pyrimidine containing nnrti with rt inhibitors - Google Patents

Abstract

The present invention relates to NNRTI, designated TMC278, useful for the treatment of HIV-infected patients or for preventing HIV transmission or infection, as nucleoside reverse transcriptase inhibitors such as emtricitabine, lamivudine or abakavir and / or nucleotide reverse transcriptase inhibitors. And to pyrimidine blends comprising tenofovir.

Description

Combination of pyrimidine-containing NNTRi and RT inhibitors {COMBINATIONS OF A PYRIMIDINE CONTAINING NNRTI WITH RT INHIBITORS}

The present invention relates to pyrimidine combinations comprising NNRTI with nucleoside reverse transcriptase inhibitors and / or nucleotide reverse transcriptase inhibitors useful for the treatment of HIV-infected patients or for preventing HIV transmission or infection.

Although markedly advanced by the introduction of HAART therapy (Highly Active Anti-Retroviral Therapy), nucleoside reverse transcriptase inhibitors (NRTIs), non-nucleoside reverse transcriptase inhibitors ( Resistance of the HIV virus to NNRTIs), nucleotide reverse transcriptase inhibitors (NtRTIs), protease inhibitors, and recent fusion inhibitors is a major cause of therapy failure. For example, half of patients who receive anti-HIV combination therapy because of the resistance of the virus to one or more drugs used do not respond to the treatment as a whole. In addition, resistant viruses can affect even newly infected individuals, resulting in severe limited therapeutic options for patients who have not received the drug. At an international AIDS conference in Paris in July 2003, the researchers found that the largest study of AIDS drugs found that about 10% of all newly infected patients in Europe had drug-resistant strains. Smaller tests were conducted to measure immunity propagation at high-risk city centers in San Francisco. The test showed that the highest level of resistance was 27%.

The pharmacokinetic profiles of many commercially available antiviral agents do not allow relatively low therapeutic doses. Antiviral agents with low solubility and low pharmacokinetic profiles cause AIDS patients to face large amounts of pill burden, particularly those not receiving drugs or undesirable for first line therapy. In addition, as a result of AIDS viruses that are resistant to antiviral combination therapy, doctors will increase the plasma levels of the active drug in order to allow the antiviral agent to restore efficacy against mutated HIV viruses, resulting in pill burden. Will increase even more. Increasing plasma levels may increase the risk of incompatibility with prescription therapy and increase side effects.

To date, there have been several attempts to devise combination therapies. For example, a combination of 150 mg dose of lamivudine (nucleoside RT inhibitor, or 3TC) and 300 mg dose of zidovudine (nucleotide RT inhibitor, or AZT) is formulated as an oral tablet and administered twice daily, or 300 A combination of mg abakavir (nucleoside RT inhibitor) and the same amount of abakavir sulfate, 150 mg lamivudine and 300 mg dozibudine is formulated as an oral tablet and administered twice daily.

WO 93/23021 describes therapeutic combinations for the treatment of HIV-infectious agents comprising zidovudine and agents that enhance the activity of antiviral agents against HIV populations resistant to zidovudine.

WO 96/01110 describes triple combinations of rojilides, which are zidovudine, lamivudine and non-nucleoside RT inhibitors of the α-APA class.

For an overview of novel antiretroviral drugs, see Clinical Microbiology and Infection 2003, Vol. 9: 3, pp. 186-193.

WO 03/016306 specifically describes more than 250 pyrimidine derivatives having HIV replication inhibitory properties which act as non-nucleoside RT inhibitors (NNRTIs) having the ability to inhibit replication of wild type and mutant strains. One of the NNRTIs is 4-[[4-[[4- (2-cyanoethenyl) -2,6-dimethylphenyl] -amino] -2-pyrimidinyl] amino] -benzonitrile (hereinafter TMC278) to be. WO 03/016306 also describes a method for synthesizing these compounds. In addition, other antiviral agents such as suramin, pentamidine, thymopentin, castanospermine, dextran (dextran sulfate), foscameth-sodium (trisodium phosphono formate), zidobudine (3'-azide) Fig. 3'-deoxythymidine, AZT), didanosine (2 ', 3'-di-deoxyinosine; ddI), zalcitabine (dideoxytidine, ddC), lamivudine (2'-3'- Dideoxy-3'-thiacytidine, 3TC), stavudine (2 ', 3'-didehydro-3'-deoxythymidine, d4T), abacavir, nevirapine (11-cyclopropyl-5, 11 -Dihydro-4-methyl-6H-dipyrido- [3,2-b: 2 ', 3'-e] [1,4] diazepin-6-one), efavirens, deravirdin, TMC120 , TMC125, tenofovir, (S) -8-chloro-4, 5,6,7-tetrahydro-5-methyl-6- (3-methyl-2-butenyl) imidazo- [4,5, 1-jk] [1,4] benzodiazepine-2 (1W) -thione, α-[(2-nitrophenyl) amino] -2,6-dichloro-benzene-acetamide, RO-5-3335, indinavir , Ritonavir, saquinavir, lopinavir (ABT-37 8), Nelpinavir, Amprenavir, TMC126, BMS-232632, VX-175, T-20, T-1249, AMD-3100 and a combination of hydroxyurea and NNRTI are described.

Despite current combination therapies, there is a need for improved antiviral therapy, more particularly AIDS therapy. There is a particular need for therapy that is effective against wild type HIV viruses, and the more common resistant HIV virus. It may be particularly desirable for first line therapy to limit or inhibit the recurrence of drug resistant viruses and to devise combination therapy with a low burden of pills, which can be used for a long time and remain long term.

It is an object of the present invention to provide a combination of one or more therapeutically effective antiviral drugs that can be used as first line therapy in patients who have not been administered the drug for a long time.

It is an object of the present invention to provide a method for the treatment of one or more therapeutically effective antiviral drugs, in which the antiviral drug has a complementary resistance profile, allowing patients with high barriers of resistance and who have not received the drug to take the formulation for a long time. To provide a formulation.

Another object of the present invention is to provide a combination of one or more therapeutically effective antiviral drugs in which each active antiviral drug in the combination is administered once daily to reduce the patient's pill burden.

A further object of the present invention is to provide a combination of one or more therapeutically effective antiviral drugs in which each active antiviral drug in the combination can be formulated together.

It is a further object of the present invention to provide a combination of one or more therapeutically effective antiviral drugs in which each active antiviral drug in a therapeutically effective amount of the combination can be formulated together in one single pharmaceutical formulation.

It is another object of the present invention to provide a combination of one or more therapeutically effective antiviral drugs that can be used to prevent HIV transmission or infection in humans.

All references cited herein are incorporated by reference.

Accordingly, according to the first aspect the present invention provides a composition comprising (i) TMC278 or a stereoisomeric form thereof; Or a pharmaceutically acceptable salt thereof; Or prodrugs thereof; And (ii) nucleoside reverse transcriptase inhibitors and / or nucleotide reverse transcriptase inhibitors; TMC278 and nucleotide reverse transcriptase inhibitors and / or nucleoside reverse transcriptase inhibitors provide combinations that are therapeutically effective HIV inhibitors at doses that can be administered once daily.

Accordingly, according to a second aspect, the present invention provides a kit comprising (i) the stereoisomeric form of TMC278; Or a pharmaceutically acceptable salt thereof; Or prodrugs thereof; And (ii) nucleoside reverse transcriptase inhibitors; TMC278 and nucleoside reverse transcriptase inhibitors provide combinations that are therapeutically effective HIV inhibitors at doses that can be administered once daily.

According to a third aspect, the present invention provides (i) a stereoisomeric form of TMC278; Or a pharmaceutically acceptable salt thereof; Or prodrugs thereof; And (ii) nucleotide reverse transcriptase inhibitors; TMC278 and nucleoside reverse transcriptase inhibitors provide combinations that are therapeutically effective HIV inhibitors at doses that can be administered once daily.

According to a fourth aspect, the present invention provides a composition comprising (i) TMC278 in its stereoisomeric form; Or a pharmaceutically acceptable salt thereof; Or prodrugs thereof; And (ii) nucleoside reverse transcriptase inhibitors; And (iii) nucleotide reverse transcriptase inhibitors; TMC278 and nucleoside reverse transcriptase inhibitors and nucleotide reverse transcriptase inhibitors provide triple combinations that are therapeutically effective HIV inhibitors at doses that can be administered once daily.

According to a fifth aspect, the present invention provides a composition comprising (i) TMC278 in its stereoisomeric form; Or a pharmaceutically acceptable salt thereof; Or prodrugs thereof; And (ii) nucleoside reverse transcriptase inhibitors; And (iii) a second nucleoside reverse transcriptase inhibitor that is different from the nucleoside reverse transcriptase inhibitor of (ii); TMC278 and the first and second nucleoside reverse transcriptase inhibitors provide triple combinations that are therapeutically effective HIV inhibitors at doses that can be administered once daily.

According to yet another aspect there is provided a pharmacological preparation comprising a pharmaceutically acceptable carrier and a combination described herein.

The invention also relates to the use of the combinations described herein as HIV inhibitors and to their use in the treatment of HIV infected patients or in the prevention of HIV transmission or infection.

The present invention is based on the discovery that TMC278 is an potent reverse transcriptase inhibitor with an extremely high genetic barrier with a desirable pharmacokinetic profile that allows once-daily administration. It is surprising to discover that the TMC278 has all these properties. What mutations in the HIV-1 genome are screened by the given drug, whether the mutated virus can have a chance of survival under the pressure of the drug, and how much drug is needed to limit or inhibit the recurrence of the mutated virus This is characteristic because it is impossible to predict how often the drug should be provided to maintain the suppression of the development of resistant viruses that can overcome the genetic barrier of the drug.

As used herein, the term 'therapeutically effective HIV inhibitor at a dose that can be administered once daily' means that the HIV inhibitor is suitable for administration every 24 hours. The term 'appropriate for administration every 24 hours' means that the HIV inhibitor is administered every 24 hours and is effective in inhibiting HIV infection over 24 hours by providing the active ingredient in an effective plasma concentration. HIV inhibitors for use in the present invention may be administered every 24 hours.

TMC278 or 4-[[4-[[4- (2-cyanoethenyl) -2, 6-dimethylphenyl] -amino] -2-pyrimidinyl] -amino] -benzonitrile is known as W003 / It may be prepared as described in 016306. TMC278 can be used in base form or, preferably, in suitable pharmaceutically acceptable salt form, in particular as acid addition salt form. Pharmaceutically acceptable addition salts are meant to include therapeutically active non-toxic salt forms. Acid addition salt forms include base acid salts, such as inorganic acids, such as hydrochloric acid, such as hydrochloric or hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and pseudoacids; Or organic acids, such as acetic acid, propanoic acid, hydroxyacetic acid, 2-hydroxypropanoic acid, 2-oxopropanoic acid, oxalic acid, malonic acid, succinic acid, maleic acid, fumaric acid, malic acid, tartaric acid, 2-hydroxy-1 With 2,3-propanetricarboxylic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, 4-methylbenzenesulfonic acid, cyclohexanesulfonic acid, 2-hydroxybenzoic acid, 4-amino-2-hydroxybenzoic acid, and the like. Can be obtained by treatment. Hydrogen halide salts, in particular hydrochloride salts, are preferred for use in the present invention.

TMC278 is formed in stereoisomeric forms, more particularly in E- and Z-isomeric forms. Both isomers can be used in the combinations of the present invention. When referred to in the context of TMC278, it is meant to include both E- and Z- and mixtures of both forms.

Preferred forms of TMC278 for use in the present invention are E-isomers, namely (E) -4-[[4-[[4- (2-cyanoethenyl) -2,6-dimethylphenyl] -amino] -2 -Pyrimidinyl] -amino] -benzonitrile (hereinafter referred to as E-TMC278). Z-isomer of TMC278, ie (Z) -4-[[4-[[4- (2-cyanoethenyl) -2, 6-dimethylphenyl] -amino] -2-pyrimidinyl] -amino] -Benzonitrile (hereinafter referred to as compound Z-TMC278) can also be used. It has a relatively high potency against nocturnal HIV-1 but less activity against single and double mutants compared to E-isomers. Table 1 shows the IC ₅₀ values (nM) of the E and Z isomers of TMC278.

Table 1

When referring to the E-form of TMC278 (ie, E-TMC278), it is meant to include isomers of the E- and Z-form in which the pure E-isomer or E-form predominantly exists, ie Isomeric mixtures comprising at least 50% or in particular at least 80% of the E-form, or at least 90% of the E-form. Of particular interest are the E-forms, which are substantially free of Z-forms. Substantially free in this context refers to an isomeric mixture comprising an E-Z-mixture with no or little Z-form, eg, 90%, in particular 95% or 98% or 99%. Equally, when referring to the Z-form of TMC278 (ie, Z-TMC278), it is meant to include mixtures of Z- and E-forms in which the pure Z- or Z-form is predominantly present. That is, isomeric mixtures comprising at least 50% or in particular at least 80% Z-form, or at least 90% Z-form. Of particular interest is the Z-form, which is substantially free of the E-form. Substantially free in this context refers to an isomeric mixture comprising an E-Z-mixture with little or no E-form, such as 90%, in particular 95% or 98% or 99% Z-form. Salts in the isomeric form of TMC278, in particular the salts mentioned above, are meant to be included for use in the present invention. Of particular interest are Z-TMC278 hydrochloride and especially E-TMC278 hydrochloride.

Advantageously, the nucleotide reverse transcriptase inhibitor and nucleoside reverse transcriptase inhibitor select mutations in the reverse transcriptase that do not cause resistance to TMC278. Of particular interest are (1) TMC278 and nucleoside / nucleotide reverse transcriptase inhibitors or therapeutically effective HIV inhibitors at doses that can be administered once daily (2) nucleoside / nucleotide reverse transcription Enzyme inhibitors or inhibitors are combinations described herein that select for mutations in reverse transcriptases that do not cause resistance to TMC278.

In particular, in one aspect, comprising (i) TMC278 or its stereoisomeric form or pharmaceutically acceptable salt or prodrug thereof, and (ii) nucleoside reverse transcriptase inhibitors, comprising (1) TMC278 and nucleoside / nucleotides Reverse transcriptase inhibitors or inhibitors are therapeutically effective HIV inhibitors at doses that can be administered once daily, and (2) nucleoside reverse transcriptase inhibitors or inhibitors select mutations in the reverse transcriptase that do not cause resistance to TMC278. To provide a blend. In another aspect, (i) TMC278 or its stereoisomeric form or pharmaceutically acceptable salt or prodrug thereof, and (ii) nucleoside reverse transcriptase inhibitors, comprising (1) TMC278 and nucleoside / nucleotides A combination wherein the reverse transcriptase inhibitor or inhibitors are therapeutically effective HIV inhibitors at doses that can be administered once daily and (2) nucleotide reverse transcriptase inhibitors or inhibitors select mutations in the reverse transcriptase that do not cause resistance to TMC278. To provide.

In a preferred embodiment, it comprises (i) TMC278 or its stereoisomeric form or pharmaceutically acceptable salt or prodrug thereof, and (ii) nucleoside reverse transcriptase inhibitor, and (iii) nucleotide reverse transcriptase inhibitor, TMC278 and nucleotide reverse transcriptase inhibitors and nucleoside reverse transcriptase inhibitors are therapeutically effective HIV inhibitors at doses that can be administered once daily (2) nucleotide reverse transcriptase inhibitors and nucleoside reverse transcriptase inhibitors to TMC278 Triple combinations are provided that select mutations in reverse transcriptases that do not cause resistance. In another preferred aspect, (i) TMC278 or its stereoisomeric form or a pharmaceutically acceptable salt or prodrug thereof, and (ii) nucleoside reverse transcriptase inhibitors, and (iii) the nucleoside reverse transcriptase of (ii) A second nucleoside reverse transcriptase inhibitor that is different from the inhibitor; (1) TMC278 and nucleoside reverse transcriptase inhibitors are therapeutically effective HIV inhibitors at doses that can be administered once daily and (2) nucleoside reverse transcriptase inhibitors are used in reverse transcriptase that do not cause resistance to TMC278. Triple combinations are provided for selecting mutations.

Preferred nucleotide reverse transcriptase inhibitors that can be used in the combinations of the present invention include tenofovir and its prodrug tenofovir disoproxyl fumarate.

Tenofovir is a currently available adenosine nucleotide analogue having activity against retroviruses. Tenofovir disoproxyl fumarate (tenofovir DF) is an orally administrable prodrug of disposable tenofovir per day. For antiviral activity, tenofovir DF needs to be hydrolyzed to ANP analogs and phosphorylated to active diphosphate sites [Arimilli et al Antiviral Chemistry and Chemotherpy 1997, 8: 6 (557-564); Fridland et al. Antiviral Research 1997, 34]. After invading into lymphocytes or macrophages, the prodrug is converted into parent analogs, tenofovir, and phosphorylated into mono- and diphosphate metabolites. Cellular enzymes responsible for phosphorylation of the drug are adenylate kinase and nucleoside diphosphate kinase [Robbins et al. Antimicrobial Agents and Chemotherapy 1995, 39: 10 (2304-2308); Robbins et al. Antimicrobial Agents and Chemotherapy 1998, 42: 3 (612-617). Unlike other nucleoside analogs, such as zidovudine or stavudine, whose phosphorylation is dependent on the cell cycle, tenofovir is efficiently phosphorylated when stabilizing and cycling peripheral blood lymphocytes [Robbins et al. 1998]. Tenofovir can inhibit HIV-1 replication in different cell types that can target HIV, including primary human blood lymphocytes and macrophages [Perno et al. Antiviral Research 1992 (289-304); Perno et al. Molecular Pharmacology 1996, 50: 2 (359-366)]. The first target of tenofovir diphosphate is reverse transcriptase (RT). Tenofovir diphosphate is a competitive inhibitor for incorporating deoxyadenosine triphosphate into neonatal proviral DNA chains. Inhibition of HIV-1 RT by tenofovir diphosphate has an inhibition constant of approximately 0.9 μM and can terminate additional chain elongation when the analog is incorporated into the growing viral DNA chain. Tenofovir inhibits viral RT even more effectively than inhibiting cellular DNA polymerase [Suo et al Journal of Biological Chemistry 1998,273: 42 (2750-2758)]. The concentration (EC ₅₀ ) required to inhibit replication of various HIV-1 strains by 50% in lymphocyte and magrophage cell types (MT-2, CEM, ACH8) ranges from 0.2 to 10 μΜ. Antiviral effects are achieved at nontoxic doses of tenofovir (optionally in the range of 100 to 2000). Tenofovir DF is currently available as a 300 mg tablet taken once daily.

Viral resistance to tenofovir slowly appears in vitro. Recombinant viruses expressing K65R mutations exhibited a three-fold reduced sensitivity in vitro [Cherrington et al. Interscience Conference on Antimicrobial Agents and Chemo-Therapy 1997, 37th]. In particular, clinical HVI strains expressing M184V rubivudine-associated resistance mutations on RT, regardless of changes to mutant enzymes (Ki), showed increased sensitivity to tenofovir in wild-type or in vitro [Miller et al. . Interscience Conference on Antimicrobial Agents and Chemotherapy 1998,]. Long-term treatment (5-15 weeks) of rhesus macaque monkeys born with tenofovir (dosage 30 mg / kg), starting at 3 weeks after inoculation with monkey immunodeficiency virus, results in susceptibility to tenofovir. An SIV emerged, which was approximately 5-fold reduced [Van Rompay et al. Antimicrobial Agents and Chemotherapy 1996, 40: 11 (2586-2591). Such low levels of resistance are associated with the appearance of K65R mutations.

In a preferred embodiment, it comprises (i) TMC278 or its stereoisomeric form or pharmaceutically acceptable salt or prodrug thereof, and (ii) tenofovir or its prodrug tenofovir disoproxyl fumarate, including TMC278 and Tenofovir or its prodrug Tenofovir disoproxyl fumarate provides a combination that is a therapeutically effective HIV inhibitor in a dose that can be administered once daily.

In another example, (i) TMC278 or its stereoisomeric form or a pharmaceutically acceptable salt or prodrug thereof, and (ii) nucleoside reverse transcriptase inhibitors, and (iii) tenofovir disoproxyl fumarate Including, but not limited to, TMC278 and nucleoside reverse transcriptase inhibitors and tenofovir disoproxyl fumarate provide triple combinations of therapeutically effective HIV inhibitors that can be administered once daily.

Preferred nucleoside reverse transcriptase inhibitors that can be used in the combinations of the present invention include abakavir or pharmaceutically acceptable salts thereof, emtricitabine, racemic FTC and lumivudine (or 3TC).

Emtricitabine or (-)-FTC is racemic FTC or (±) -cis-4-amino-5-fluoro-1- [2- (hydroxymethyl) -1,3-oxathiolan-5-yl ] -2 (1H) -pyrimidinone (FTC) is the left (-) rotary enantiomer form. It is a commercially available nucleoside analogue and has been shown to be active against HIV-1 [Hoong et al. Journal of Organic Chemistry 1992 (5563-5565); Jeong et al Journal of Medicinal Chemistry 1993, 36: 2 (181-195); Van Roey et al. Antiviral Chemistry and Chemotherapy 1993,4: 6 (369-375) .The in vitro anti-HIV-1 activity of FTC's (-)-beta enantiomer is at least 20 times that of (+)-beta-enantiomer. (+)-Enantiomers have been reported to be significantly more toxic to bone marrow precursor cells than (-)-enantiomers (Schinazi et al Antimicrobial Agents and Chemotherapy 1992, 36: 11 (2423-2431)). Successive passages of the virus in human PBMCs and MT-2 cells with increasing drug concentration assessed the efficacy of HIV-1 resistance to FTC.High drug-resistant HIV-1 variants were found after two or more infection cycles. Replication of virus populations was dominated RT derived from drug-resistant virus particles was 15- to 50-fold less susceptible to 5'-triphosphate of FTC compared to enzymes from parent drug sensitive virus. RT gene DNA sequencing confirmed mutations from Met (ATG) to Val (GTG or GTA) at the 184th codon [Schinazi etal Antimicrobial Agents and Chemotherapy 1993,37: 4 (875-881); Tisdale et al Antiviral Research 1993,20: Suppl 1; Smith et al Journal of Virology 1997,71: 3 (2357-2362); Harrer et al Jouynal of infectious Diseases 1996,173: 2 (476-479); Tisdale et al Proceedings of the National Academy of Sciences of the United States of America 1993, 90: 12 (5653-5656)] Since no single mutation was observed in YMDD of reverse transcriptase in HIV-infected patients, (-)-FTC is not appropriate for monotherapy and is effective in treating HIV-infected patients. Should be administered in combination with other antiretroviral agents to be treated. Emtricitabine can be used as a 200 mg capsule administered once daily.

The chemical name of Lumivudine is (-)-2 ', 3'-dideoxy-3'-thiacytidine, and is described, for example, as an antiviral nucleoside analogue in EP-382 526. Also, by way of example, it is a well established and useful antiretroviral agent commercially available as a 150 mg oral tablet. Lumivudine is formulated with zidovudine (300 mg zidovudine / 150 mg lumivudine) and commercially available in combination with lumivudine and abakavir sulfate (300 mg zidovudine / 150 mg lumivudine / equivalent to 300 mg abacavir).

Abacavir is a well-established and useful commercially available oral solution of abacavir sulfate at a concentration equivalent to 20 mg abakavir base / ml, or as an oral tablet of abacavir sulfate at a concentration equivalent to 300 mg abakavir base. Antiretroviral agent. Abacavir sulfate is also commercially available in combination with luminubidine and zidovudine (300 mg dozivudine / 150 mg lumivudine / equivalent to 300 mg abakavir).

Abakavir is a carbocyclic nucleoside with potent and selective anti-HIV activity. Abacavir in its optically active form is described in EP-434 450. The cis-isomer of Abakavir with non-specific absolute stereochemical configuration is described in EP-349 242. Abakavir is one of the most potent NRTIs currently developed. After short-term abacavir monotherapy, an average viral load of 1.4 log10 RNA copies / ml was observed to decrease. In vitro, resistant viruses are not rapidly selected by Abakavir. No significant decrease in sensitivity to abakavir was observed in wild-type or zidovudine-resistant HIV-1 strains until after 8-10 passages in MT-4 cells. One set of resistance mutations at HIV reverse transcriptase (RT) codons, 65R, 74V, 115F, and / or 184V are selected for in vitro passage using abakavir, and the combination of these mutations is selected for Abakavir in laboratory HIV strains. It was necessary to reduce the sensitivity to 10 times. The first mutation detected when subcultured HIV-1 with increasing concentrations of abakavir is M184V, which only reduces the sensitivity to HIV-1 only three-fold. Phenotypic resistance to 3TC and / or the presence of M184V mutations does not prevent the viral load response to abakavir therapy. Resistance to multiple nucleosides is associated with reducing or eliminating resistance to abakavir [Kumar et al Antimicrobial Agents and Chemotherapy 1999, 43: 3 (603-608); Lanier et al International Conference on Retro Viruses and Opportunistic Infections 1998, 5th: Chicago; posted on 16 April 1999].

In a preferred example, (i) TMC278 stereoisomeric form thereof; Or a pharmaceutically acceptable salt thereof; Or a prodrug thereof, and (ii) emtricitabine, wherein TMC278 and emtricitabine provide a combination that is a therapeutically effective HIV inhibitor in a dose that can be administered once daily.

In a preferred example, (i) TMC278 stereoisomeric form thereof; Or a pharmaceutically acceptable salt thereof; Or prodrugs thereof; And (ii) luminubidine, wherein TMC278 and luminubidine provide a combination that is a therapeutically effective HIV inhibitor in a dose that can be administered once daily.

In another example, (i) TMC278 stereoisomeric form thereof; Or a pharmaceutically acceptable salt thereof; Or prodrugs thereof; And (ii) abakavir or a pharmaceutically acceptable salt thereof, wherein TMC278 and abakavir are doses of a therapeutically effective HIV inhibitor that can be administered once daily.

In another example, (i) TMC278 stereoisomeric form thereof; Or a pharmaceutically acceptable salt thereof; Or prodrugs thereof; And (ii) abakavir sulfate, wherein TMC278 and abakavir sulfate are doses of therapeutically effective HIV inhibitors that can be administered once daily.

In another example, (i) TMC278 stereoisomeric form thereof; Or a pharmaceutically acceptable salt thereof; Or prodrugs thereof; And (ii) emtricitabine, and (iii) nucleotide reverse transcriptase inhibitor, wherein TMC278 and nucleoside reverse transcriptase inhibitor and emtricitabine are triple doses of therapeutically effective HIV inhibitors that can be administered once daily Provide the formulation.

In another example, (i) TMC278 or its stereoisomeric form; Or a pharmaceutically acceptable salt thereof; Or prodrugs thereof; And (ii) luminubidine, and (iii) nucleotide reverse transcriptase inhibitors, wherein TMC278 and nucleoside reverse transcriptase inhibitors and lumivudine are triple doses of therapeutically effective HIV inhibitors that can be administered once daily To provide.

In another example, (i) TMC278 or its stereoisomeric form; Or a pharmaceutically acceptable salt thereof; Or prodrugs thereof; And (ii) abakavir or a pharmaceutically acceptable salt thereof, or preferably abakavir sulfate, and (iii) a nucleotide reverse transcriptase inhibitor, wherein the TMC278 and nucleoside reverse transcriptase inhibitor and abakavir or a pharmaceutically acceptable salt thereof Acceptable salts, or preferably abakavir sulfate, provide triple combinations that are therapeutically effective HIV inhibitors in doses that can be administered once daily.

In another example, (i) TMC278 or its stereoisomeric form; Or a pharmaceutically acceptable salt thereof; Or prodrugs thereof; And (ii) emtricitabine, and (iii) tenofovir or prodrug tenofovir disoproxyl fumarate, including TMC278 and emtricitabine and tenofovir or prodrug tenofovir disoproxyl Fumarate provides triple combinations that are therapeutically effective HIV inhibitors in doses that can be administered once daily.

In another example, (i) TMC278 stereoisomeric form thereof; Or a pharmaceutically acceptable salt thereof; Or prodrugs thereof; And (ii) luminubidine and (iii) tenofovir or prodrug tenofovir disoproxyl fumarate, wherein TMC278 and luminubidine and tenofovir or its prodrug tenofovir disoproxyl fumarate Provided are triple combinations of therapeutically effective HIV inhibitors that can be administered once daily.

In another example, (i) TMC278 stereoisomeric form thereof; Or a pharmaceutically acceptable salt thereof; Or prodrugs thereof; And (ii) abakavir or a pharmaceutically acceptable salt form thereof, preferably abacavir sulfate; And (iii) tenofovir or prodrug thereof tenofovir disoproxyl fumarate, including TMC278 and abakavir or a pharmaceutically acceptable salt form thereof, preferably abacavir sulfate and tenofovir or a prodrug thereof Tenofovir disoproxyl fumarate provides a triple combination of therapeutically effective HIV inhibitors in a dose that can be administered once daily.

Preferred triple combinations also include:

(a) TMC278 stereoisomeric forms; Or a pharmaceutically acceptable salt thereof; Or prodrugs thereof; And emtricitabine and tenofovir disoproxyl fumarate;

(b) TMC278 stereoisomeric forms; Or a pharmaceutically acceptable salt thereof; Or prodrugs thereof; And lumivudine and tenofovir disoproxyl fumarate.

(c) TMC278 stereoisomeric forms; Or a pharmaceutically acceptable salt thereof; Or prodrugs thereof; And abacavir sulfate and tenofovir disoproxyl fumarate.

(d) TMC278 stereoisomeric forms; Or a pharmaceutically acceptable salt thereof; Or prodrugs thereof; And emtricitabine and lumivudine;

(e) TMC278 stereoisomeric forms; Or a pharmaceutically acceptable salt thereof; Or prodrugs thereof; Emtricitabine and abakavir or a pharmaceutically acceptable salt thereof, preferably abacavir sulfate.

(f) TMC278 stereoisomeric form; Or a pharmaceutically acceptable salt thereof; Or prodrugs thereof; Abacavir or a pharmaceutically acceptable salt thereof, preferably abacavir sulfate and luminvudine.

In particular, in each combination (a)-(f) the active ingredient, in particular TMC278, emtricitabine, lumivudine, abacavir or a pharmaceutically acceptable salt form thereof, preferably abacavir sulfate, and tenofovir or its Prodrug tenofovir disoproxyl fumarate is a therapeutically effective HIV inhibitor in a dose that can be administered once daily.

The dual combinations of the present invention may comprise one or more additional active ingredients, which may be agents or other active ingredients useful for the treatment of HIV infected patients. The triple combination of the present invention may comprise one or more additional active ingredients which may equally be a medicament or other active ingredient useful for treatment in an HIV infected patient. Preferably, the additional agents are therapeutically effective HIV inhibitors in a dose that can be administered once daily.

The active ingredients in the combinations of the invention may be administered simultaneously, concurrently, or sequentially. Simultaneous administration may be performed using a single pharmaceutical agent or separate pharmaceutical agents. Generally, the combination may be administered via topical, oral, rectal, intravenous, subcutaneous or intramuscular routes.

Co-administration using a single pharmaceutical agent is preferred for the first therapy for HIV infection.

Thus, in another aspect, there is provided a product comprising a formulation as described herein as a formulation for simultaneous, separate or sequential use against HIV infection.

The invention also provides combinations for simultaneous, separate or sequential use against HIV infection in the form of (i) TMC278 stereoisomeric forms thereof; Or a pharmaceutically acceptable salt thereof; Or prodrugs thereof; And (ii) a nucleoside reverse transcriptase inhibitor and / or nucleotide reverse transcriptase inhibitor; TMC278 and nucleoside reverse transcriptase inhibitors and / or nucleoside reverse transcriptase inhibitors provide products that are therapeutically effective HIV inhibitors in doses that can be administered once daily.

In a further aspect, as a combination for simultaneous, separate or sequential use against HIV infection (i) TMC278 stereoisomeric form thereof; Or a pharmaceutically acceptable salt thereof; Or prodrugs thereof; And (ii) nucleoside reverse transcriptase inhibitors; TMC278 and nucleoside reverse transcriptase inhibitors provide products that are therapeutically effective HIV inhibitors in doses that can be administered once daily.

In another aspect, as a combination for simultaneous, separate or sequential use against HIV infection (i) TMC278 stereoisomeric form thereof; Or a pharmaceutically acceptable salt thereof; Or prodrugs thereof; And (ii) nucleotide reverse transcriptase inhibitors, wherein the TMC278 and nucleoside reverse transcriptase inhibitors provide a product that is a therapeutically effective HIV inhibitor that can be administered once daily.

In another aspect, there is provided a combination for simultaneous, separate or sequential use against HIV infection in (i) TMC278 or a stereoisomeric form thereof; Or a pharmaceutically acceptable salt thereof; Or prodrugs thereof; And (ii) nucleoside reverse transcriptase inhibitors; And (iii) nucleotide reverse transcriptase inhibitors; TMC278 and nucleotide reverse transcriptase inhibitors and nucleoside reverse transcriptase inhibitors provide products that are therapeutically effective HIV inhibitors in doses that can be administered once daily.

In another aspect, as a combination for simultaneous, separate or sequential use against HIV infection (i) TMC278 stereoisomeric form thereof; Or a pharmaceutically acceptable salt thereof; Or prodrugs thereof; And (ii) nucleoside reverse transcriptase inhibitors; And (iii) a second nucleoside reverse transcriptase inhibitor that is different from the nucleoside reverse transcriptase inhibitor of (ii); TMC278 and nucleoside reverse transcriptase inhibitors provide products that are therapeutically effective HIV inhibitors in doses that can be administered once daily.

In the product of the present invention, the active ingredient is present in a therapeutically effective amount, wherein a therapeutically effective amount means an amount sufficient to exert a sufficient HIV inhibitory effect for a certain period of time, ie for a time interval when taking each formulation, preferably about 24 hours. do.

A product as described above comprising one or more specific active ingredients as described herein, such as emtricitabine, racemic FTC, lamivudine, tenofovir and prodrug tenofovir disoproxyl fumarate thereof It is a preferable example.

The above-mentioned products may comprise separate formulations of the active ingredient or, when applied, may formulate two or more active ingredients together.

In a further aspect the present invention provides a pharmaceutical formulation comprising a combination as described herein and a suitable carrier.

In another aspect, there is provided a pharmaceutical composition comprising (i) TMC278 a stereoisomeric form thereof as a pharmaceutically acceptable carrier and active ingredient; Or a pharmaceutically acceptable salt thereof; Or prodrugs thereof; And (ii) a nucleoside reverse transcriptase inhibitor and / or nucleotide reverse transcriptase inhibitor; TMC278 and nucleoside reverse transcriptase inhibitors and / or nucleoside reverse transcriptase inhibitors provide pharmaceutical preparations that are therapeutically effective HIV inhibitors at doses that can be administered once daily.

In another aspect, the present invention further provides pharmaceutically acceptable carriers and active ingredients as (i) TMC278 stereoisomeric forms; Or a pharmaceutically acceptable salt thereof; Or prodrugs thereof; And (ii) nucleoside reverse transcriptase inhibitors; TMC278 and nucleoside reverse transcriptase inhibitors provide pharmaceutical preparations that are therapeutically effective HIV inhibitors at doses that can be administered once daily.

In yet another aspect, the present invention further provides pharmaceutically acceptable carriers and active ingredients as (i) stereomeric forms of TMC278; Or a pharmaceutically acceptable salt thereof; Or prodrugs thereof; And (ii) a nucleotide reverse transcriptase inhibitor, wherein the TMC278 and nucleoside reverse transcriptase inhibitor and nucleoside reverse transcriptase inhibitor are therapeutically effective HIV inhibitors at a dose that can be administered once daily. do.

In yet another aspect, the present invention further provides pharmaceutically acceptable carriers and active ingredients as (i) stereomeric forms of TMC278; Or a pharmaceutically acceptable salt thereof; Or prodrugs thereof; And (ii) nucleoside reverse transcriptase inhibitors; And (iii) nucleotide reverse transcriptase inhibitors; TMC278 and nucleoside reverse transcriptase inhibitors and nucleoside reverse transcriptase inhibitors provide pharmaceutical formulations that are therapeutically effective HIV inhibitors at doses that can be administered once daily.

In yet another aspect, the present invention further provides pharmaceutically acceptable carriers and active ingredients as (i) stereomeric forms of TMC278; Or a pharmaceutically acceptable salt thereof; Or prodrugs thereof; And (ii) nucleoside reverse transcriptase inhibitors; And (iii) a second nucleoside reverse transcriptase inhibitor that is different from the nucleoside reverse transcriptase inhibitor of (ii); TMC278 and nucleoside reverse transcriptase inhibitors provide pharmaceutical preparations that are therapeutically effective HIV inhibitors at doses that can be administered once daily.

In the pharmaceutical formulations of the present invention, the active ingredient is administered in a therapeutically effective amount, the therapeutically effective amount being an amount sufficient to exert sufficient HIV inhibitory effect for a certain period of time, i. Means.

A product as described above comprising one or more specific active ingredients as described herein, such as emtricitabine, racemic FTC, lamivudine, tenofovir and prodrug tenofovir disoproxyl fumarate thereof It is a preferable example.

The pharmaceutical formulations of the present invention may be formulated in a variety of formulations for various types of administration. In order to prepare a pharmaceutical formulation of the present invention, an effective amount of the active ingredient in the form of an addition salt is optionally combined in intimate mixture with a pharmaceutically acceptable carrier, which carrier can take various forms depending on the dosage form desired for administration. The pharmaceutical preparations of the invention are particularly formulated in unit dosage forms as appropriate for oral, rectal, transdermal or parenteral injection. For example, when preparing a formulation for oral administration,

Oral liquid preparations, such as suspending agents, syrups, exciters, emulsions and solutions; Or solid carriers such as starch, sugar, kaolin, diluents, glidants, binders, disintegrants and in the case of powders, pills, capsules and tablets, any of the usual pharmaceutical media such as water, Glycols, oils, alcohols and the like can be used. For ease of administration, tablets and tablets represent the most advantageous oral dosage unit form, in which case solid pharmaceutical carriers are used. For parenteral compositions, the carrier may also contain sterile water, at least in large amounts, although other ingredients may also be included, for example, to aid solubility. By way of example, the carrier may prepare an injectable solution comprising saline solution, glucose solution or a mixture of saline and glucose solution. Injectable suspensions may also be prepared in which appropriate liquid carriers, suspending agents and the like may be used.

In one aspect of the invention, the formulations may be formulated in oral tablet form further comprising a pharmaceutically acceptable excipient having a weight range of 150 mg to 600 mg, suitable range 200 to 400 mg.

The nominal weight range of conventional oral tablets comprising the active ingredient according to the invention is 200 mg to 1500 mg, suitably 500 mg to 1250 mg, more suitably 600 to 1100 mg.

An advantage of the pharmaceutical formulations of the present invention lies in the fact that each component of the combination can be formulated together as one pharmaceutical formulation and does not need to be administered separately. The daily therapeutic antiviral amount of the present formulation components in a single pharmaceutical formulation to be co-formulated is preferably administered in a single unit dosage form, but preferably in multiple unit dosage forms, for example 2, 3, 4, 5 It can be administered in the above unit dosage form. The physician should take into account the severity of the patient's symptoms and the patient's weight, sex and possibly other parameters, such as the rate of release, metabolism, biodistribution and absorption of each drug, and other factors known to those skilled in the art. Will determine the quantity.

The present invention also provides a method of treating an HIV infected patient comprising administering a combination as described herein.

In addition, the present invention provides nucleoside reverse transcriptase inhibitors and / or nucleotide reverse transcriptases, which can be administered once a day in therapeutically effective amounts of TMC278 and nucleoside reverse transcriptase inhibitors and / or nucleotide reverse transcriptase inhibitors. Provided is a method of treating an HIV infected patient comprising administering TMC278 or a stereoisomeric form thereof or a pharmaceutically acceptable salt or prodrug thereof in combination with an inhibitor.

In addition, the present invention provides a TMC278 or a stereoisomeric form or pharmaceutical form thereof in combination with a nucleoside reverse transcriptase inhibitor, which can be administered once a day with a therapeutically effective amount of a TMC278 and nucleoside reverse transcriptase inhibitor in the method. Provided is a method of treating an HIV infected patient, comprising administering an acceptable salt or prodrug thereof.

A further aspect of the present invention provides a TMC278 or a stereoisomeric form thereof or a pharmaceutically acceptable salt or prodrug thereof, which can be administered once a day with a therapeutically effective amount of TMC278 and a nucleotide reverse transcriptase inhibitor in the method. Provided is a method of treating an HIV infected patient comprising administering a reverse transcriptase inhibitor.

A further aspect of the invention provides nucleoside reverse transcriptase inhibitors, and nucleotide reverse transcriptases, which can be administered once daily with therapeutically effective amounts of TMC278, nucleoside reverse transcriptase inhibitors and nucleoside reverse transcriptase inhibitors in the method. Provided is a method of treating an HIV infected patient comprising administering TMC278 or a stereoisomeric form thereof or a pharmaceutically acceptable salt or prodrug thereof in combination with an inhibitor.

A further aspect of the invention provides an agent different from the nucleoside reverse transcriptase inhibitor, and a nucleoside reverse transcriptase inhibitor that can be administered once a day with a therapeutically effective amount of TMC278, a nucleoside reverse transcriptase inhibitor in the method. Provided is a method of treating an HIV infected patient comprising administering TMC278 or a stereoisomeric form thereof or a pharmaceutically acceptable salt or prod thereof in combination with a nucleoside reverse transcriptase inhibitor of 2.

In the method of the present invention, the active ingredient is administered in a therapeutically effective amount, which means that the therapeutically effective amount means an amount sufficient to exert sufficient HIV inhibitory effect for a certain period of time, i. do.

Preferred examples are methods as described above, wherein one or more active ingredients described herein are administered, eg, emtricitabine, racemic FTC, lamivudine, tenofovir and prodrug tenofovir disoproxyl fumarate thereof. to be.

One aspect of the invention relates to a combination of the invention for use in the manufacture of a medicament for treating an HIV infected patient.

Of particular interest are combinations as described herein, or any product, pharmaceutical formulation, unit, wherein TMC278 is E-TMC287, or preferably TMC278 hydrochloride salt, or more preferably E-TMC278 hydrochloride salt. Dosage forms, methods and uses based on such combinations.

The combinations of the present invention are particularly useful for treating AIDS and related clinical abnormalities, such as AIDS related syndromes (ARC), progressive systemic lymphadenopathy (PGL) or AIDS related nervous system abnormalities, such as multiple sclerosis. The triple combination of the present invention may be particularly useful for treating HIV-infected patients who have not received drug.

The combinations of the invention are also useful for the prevention of HIV transmission or infection, especially sexual transmission in humans. Accordingly, the present invention relates to the use of a combination of the present invention for the manufacture of a medicament for preventing HIV transmission or infection through sexual intercourse or related intimate contact between partners. The invention also relates to a method for preventing HIV transmission or infection via sexual intercourse or related intimate contact between partners comprising administering an effective amount of a combination according to the invention to a subject in need thereof.

Each daily dosage of each active ingredient in the combinations of the present invention may range from 10 mg to 800 mg, preferably 50 to 400 mg, more preferably 50 and 300 mg, or 100 to 300 mg. In particular, the daily dose of TMC278 may range from 10 mg to 500 mg, preferably 10 to 300, more preferably 50 to 250 mg, even more preferably 50 to 200 mg, for example about 100 mg. .

The weight ratio of each couple component in the triple combination taken once daily may vary from 1/10 to 10/1. Suitably, the weight ratio of each couple component varies from 1/6 to 6/1, more suitably 1/4 to 4/1, preferably 1/3 to 3/1, and more preferably 1/2 to 2 Varies from / 1.

Table 2 lists an example for the daily dose of each active ingredient in the combination of Compound E-TMC278, Emtricitabine and Tenofovir.

Table 3 lists an example for the daily dosage of each active ingredient in the combination of TMC278, Abakavir and Lumivudine (wherein the doses mentioned in the tablets for Abakavir sulfate are equivalent to Abakavir base) .

Thus, the combination of interest according to the invention is a compound E- (A) in a daily dosage in the range of 10 mg to 500 mg, luminubidine and a 300 mg equivalent daily in a daily dosage of 150 mg Dosage includes abakavir base. Suitably, the combination is formulated into a single pharmaceutical formulation.

Another interesting combination according to the invention is compound E- (A) in a daily dosage in the range of 50 mg to 250 mg, luminubidine and a 300 mg equivalent daily in a daily dosage of 150 mg Dosage includes abakavir base. Suitably, the combination is formulated into a single pharmaceutical formulation.

The invention also provides pharmaceutically acceptable carriers and active ingredients in a form adapted for application to sites where sexual intercourse or related intimate contact occurs, such as the genitals, rectum, mouth, hands, lower abdomen, upper calf, especially the vagina and mouth. A pharmaceutical composition, comprising an effective amount of the combination according to the invention. Suitable specific suitable compositions include gels, jellies, creams, ointments, films, sponges, vaginal rings, cervical caps, rectal or vaginal suppositories, vaginal or rectal or buccal tablets, oral vaginal, rectal or mouth washes And all compositions commonly used for application to the skin. To prepare pharmaceutical compositions, triple combinations of an effective amount of each specific compound as an active ingredient are closely combined with pharmaceutically acceptable carriers that can take various forms depending on the dosage form. In order to increase the residence time of the pharmaceutical composition relative to the site of administration, it may be advantageous to include a biobinding agent, in particular a biobinding polymer, in the composition. Biobinding agents may be defined as materials capable of binding to biological surfaces such as, for example, mucosal or skin tissue.

Accordingly, the present invention also relates to a pharmaceutical composition comprising an effective amount of each compound of the present invention as a active ingredient, a triple combination of the present invention and a pharmaceutically acceptable carrier, characterized in that they are bioactive to the site of application. Preferably, the site of application is the vagina, rectum, mouth or skin, most preferably the vagina.

Otten RA et al. [Journal of Virology (2000), 74 (20), 9771-9775] and Witvrouw M et al. [Antiviral Research (2000), 46 (3), 215-221] develop HIV virus after very dangerous sexual exposure. Tenofovir's ability to delay is described.

Pani A et al. (Antiviral Chemistry & Chemotherapy (2001), 12 (Suppl. 1), 51-59) described the ability of luminibudine to delay the development of the virus.

The ability of TMC278 to prevent HIV infection by sexual intercourse or related intimate contact with a partner can be demonstrated by the following test. Immature monocycle-induced branched cells (immMO-DCs) are important initiators of the lipogenic response and an excellent model for interstitial branching cells, which are early targets for sexual HIV infection. immMO-DC was used as an "in vitro" model to test the prevention of HIV infection by intercourse with a partner or related intimate contact. The model is described in the experimental section and TMC278 is described as effectively inhibiting HIV replication in MO-DC / CD4 (+) T cell coculture.

< Example >

Example  1: E- TMC278 Pharmacokinetics

To evaluate the safety, tolerability, and in vitro pharmacokinetics of a single dose of Compound E-TMC278 in healthy male volunteers, a double blind randomized placebo-modulated first trial was designed. Oral dosages of 12.5, 25, and 50 mg in PEG 400 were formulated and taken with regular food. Pharmacokinetic results are shown in Table 4.

Pharmacokinetics of a Double-blind, Randomized Placebo-Modulated First Trial with 4 Administration Periods to Assess the Safety, Tolerance, and In Vitro Pharmacokinetics of Compound E-TMC278 with a Single Oral Dose of 100 mg and 200 mg in Healthy Male Subjects The results are also reported in Table 4. During each period, six subjects were randomized to receive the same amount of Compound E-TMC278 and three subjects to placebo. The time interval between each administration period was about 14 days. Table 4 shows that a large amount of exposure and exposure proportional to the dosage were obtained. The correction factor for the five C _m data points is 0.9897 and the correction factor for the area value under the curve (AUC-48 _hr ) between 0 and 48 hours is 0.9952. The half-life range of plasma concentrations was 37 to 39 hours. Volunteers showed good resistance to the compound. No related side effects of the compound were observed.

Table 4

Example  2: compound E- TMC278 Viral profile of

Compound E-TMC278 was tested in cell-based assays using naturally occurring HIV host cells. MT-4 cells (cell lines of human T cells) were infected with HIV-1 (wild type or mutant) and exposed to different concentrations of antiviral compounds in the presence of 10% fetal calf serum. Cytotoxicity can be measured along with antiviral activity to assess the specificity of antiviral efficacy. In order to inhibit the replication step in the cell, the active compound must penetrate the cell membrane.

After 4 days incubation at 37 ° C., viability of HIV and mock-infected cells was assessed via automated tetrazolium-based colorimetric analysis. This method yielded 50% inhibitory concentration (IC50), IC90 and 50% cytotoxicity concentration (CC50) for inhibition of viral cytotoxicity. Non-CC50 / IC50, also referred to as the selection index, is an indicator of the specificity of antiviral efficacy. Wild type (wt) HIV-1 as test strain of HIV; Panels of single and double mute obtained by site-directed mutagenesis (SDM), and panels of clinical isolates selected for resistance to NNRTIs.

Wild type and SDM On mutants  Active for

A limited panel of HIV-1 mutants was constructed using specific mutagenesis (SDM) and homologous recombination techniques. Compound E-TMC278 was tested on stretched panels of single and double mutants known to be resistant to commercially available NNRTIs. Controls include nevirapine (NVP) and efavirens (EFV).

The results are shown in Table 5 (the values shown are IC ₅₀ values (nM)). For wild type virus, the IC50 obtained was 0.4 nM (0.15 ng / ml) and IC90 was 1.3 nM (0.48 ng / ml). The HIV strain with low specificity for compound E-TMC278 in this selection was a double mutant 100I ± 103N, with an IC50 of about 8 nM and an IC90 of about 16 nM.

Table 5

In vitro  Tolerance development

NNRTIs are highly selective inhibitors of HIV-1, but current clinical use is limited because of the rapid emergence of NNRTI (cross-) resistance. The incidence of resistance to compound E-TMC278 and first generation NNRTIs nevirapine and epavirens was compared in vitro.

MT4 cells with wild type HIV-1 at high multiplicity of infection (> 1 infected virus per cell, to maximize the genetic diversity of the virus community) in the presence of various concentrations of compound E-TMC278 (40,200, 1000 and 5000 x IC50) Were infected and observed twice a week for virus replication. Emerging viruses were harvested for phenotype and genotype. Cultures without evidence of viral replication were passaged for a total of 30 days in the presence of the same concentration of inhibitor (10 passages).

Resistance to nevirapine appeared within 3-6 days at all test concentrations. Alternative virus involved the usual Y181C mutation. G190E was selected at all concentrations (5 μM) within 3 to 7 days through the same experiment using Epavirens. Compound E-TMC278 failed to select resistant virus within 30 days when wild type virus was used. When double resistant mutant K103N + Y181C (IC 50 0.8 nM) was used instead of wild type virus, resistance did not appear at all test concentrations. Starting from single mutant Y181C (IC50 1.3nM) or 103N (IC50 0.3nM) did not occur at virus -breasthour 40 and 200 nM, but occurred at 10 nM.

HIV-1 resistant to first generation NNRTIs was selected very quickly in experimental settings for high gene diversity. Resistant virus involved only one mutation. In contrast, the appearance of HIV-1 resistant to Compound E-TMC278 was neither delayed nor occurred.

Compound E- TMC278 Cardiovascular and lung safety

Compound E-TMC278 has little or no effect on cardiovascular and lung parameters at in vitro concentrations including and exceeding antiviral concentrations in vivo and in vitro at and above target plasma levels in humans. Did not give.

Example  3: intercourse or related intimate contact between partners HIV  Compound E- to prevent infection Of TMC278  To test your ability In vitro  Model

For example, as a model, monocyte-induced branched cells (MO-DCs) were infected for 2 hours using monotropic HIV strain Ba-L at a multiple infectivity (MOI) of 10 ⁻³ . After infection, cells were washed six times and resuspended in 10% BCS at 400.000 cells / ml. Autologous CD4 (+) T cells were purified with the same lymphocyte fractional lavage as MO-DC and used at a concentration of 2 × 10 ⁶ cells / ml (non-MO-DC / CD4 (+) T: 1/5).

Serial dilutions of the compound of formula (I) (test compound) were added to the MO-DC / CD4 (+) T cell coculture. Each experiment was performed in a 96-well plate, each cup containing 50 μl of MO-DC, 50 μl of CD4 (+) T cells and 100 μl of test compound. 1/2 of the culture medium containing the test compound was replaced with fresh one weekly. After incubation for 14 days, the supernatant was analyzed by ELISA. In order to measure antiviral activity, the concentration of test compound (EC50) capable of inhibiting viral replication by 50% at the end of the first culture was measured. The EC 50 value for compound E-TMC278 was 0.55 nM.

Example  4:

Tablet formulations of the following composition:

Emtricitabine 300 mg

Tenofovir Diisopropyl Fumarate 300 mg

110 mg of E-TMC278 hydrochloride salt

HPMC 2910 15mPa.s 24 mg

Polysorbate 20 6 mg

Crospolividone 18 mg

Lactose Monohydrate 43 mg

Magnesium Stearate 3 mg

Talc 6 mg

The active ingredient and lactose were fluidized and sprayed with HPMC and polysorbate solution (120 ml / tablet equivalent) in water. Subsequently, crospolyvidone was added with continuous fluidization followed by magnesium stearate and talc. The granules obtained were compressed into 13 mm cylindrical tablets using a standard compaction apparatus.

Claims (25)

4-[[4-[[4- (2-cyanoethenyl) -2,6-dimethylphenyl] -amino] -2-pyrimidinyl] -amino] -benzonitrile or a stereoisomer thereof; Or a pharmaceutically acceptable salt thereof, the composition for once-daily administration for treating HIV infection. The composition of claim 1 further comprising a nucleoside reverse transcriptase inhibitor, or a nucleotide reverse transcriptase inhibitor, or a nucleoside reverse transcriptase inhibitor and a nucleotide reverse transcriptase inhibitor. 3. The method of claim 2,
(i) 4-[[4-[[4- (2-cyanoethenyl) -2,6-dimethylphenyl] -amino] -2-pyrimidinyl] -amino] -benzonitrile or a stereoisomer thereof; Or a pharmaceutically acceptable salt thereof; And
(ii) a composition comprising a nucleoside reverse transcriptase inhibitor. 3. The method of claim 2,
(i) 4-[[4-[[4- (2-cyanoethenyl) -2,6-dimethylphenyl] -amino] -2-pyrimidinyl] -amino] -benzonitrile or a stereoisomer thereof; Or a pharmaceutically acceptable salt thereof; And
(ii) a composition comprising a nucleotide reverse transcriptase inhibitor. 3. The method of claim 2,
(i) 4-[[4-[[4- (2-cyanoethenyl) -2,6-dimethylphenyl] -amino] -2-pyrimidinyl] -amino] -benzonitrile or a stereoisomer thereof; Or a pharmaceutically acceptable salt thereof;
(ii) nucleoside reverse transcriptase inhibitors; And
(iii) a composition comprising a nucleotide reverse transcriptase inhibitor. 3. The method of claim 2,
(i) 4-[[4-[[4- (2-cyanoethenyl) -2,6-dimethylphenyl] -amino] -2-pyrimidinyl] -amino] -benzonitrile or a stereoisomer thereof; Or a pharmaceutically acceptable salt thereof;
(ii) nucleoside reverse transcriptase inhibitors; And
(iii) A composition comprising a second nucleoside reverse transcriptase inhibitor other than the nucleoside reverse transcriptase inhibitor of (ii) above. The method according to claim 2, wherein the nucleotide reverse transcriptase inhibitor or nucleoside reverse transcriptase inhibitor is 4-[[4-[[4- (2-cyanoethenyl) -2,6-dimethylphenyl] -amino] -2- A composition comprising screening out mutations in reverse transcriptases that do not cause resistance to pyrimidinyl] -amino] -benzonitrile. The composition of claim 2, wherein the nucleotide reverse transcriptase inhibitor is tenofovir or a prodrug tenofovir disoproxyl fumarate thereof. The method of claim 2, wherein the nucleoside reverse transcriptase inhibitor is emtricitabine, (±) -cis-4-amino-5-fluoro-1- [2- (hydroxymethyl) -1,3-oxathiolane- 5-yl] -2 (1H) -pyrimidinone (racemic FTC), lamivudine (or 3TC), abakavir or a pharmaceutically acceptable salt thereof. The composition of claim 2, wherein the nucleoside reverse transcriptase inhibitor is emtricitabine. The composition of claim 2, wherein the nucleoside reverse transcriptase inhibitor is lamivudine. 3. The method of claim 2,
(i) 4-[[4-[[4- (2-cyanoethenyl) -2,6-dimethylphenyl] -amino] -2-pyrimidinyl] -amino] -benzonitrile or a stereoisomer thereof; Or a pharmaceutically acceptable salt thereof;
(ii) tenofovir or its prodrug tenofovir disoproxyl fumarate; And
(iii) a composition that is a tablet comprising emtricitabine. 13. The compound according to claim 12, wherein 4-[[4-[[4- (2-cyanoethenyl) -2,6-dimethylphenyl] -amino] -2-pyrimidinyl] -amino] -benzonitrile or a steric form thereof Isomers; Or 10 to 200 mg of a pharmaceutically acceptable salt thereof. The composition of claim 13 comprising 300 mg of tenofovir disoproxyl fumarate. The composition of claim 14 comprising 200 mg of emtricitabine. 9. The method of claim 8,
(i) 4-[[4-[[4- (2-cyanoethenyl) -2,6-dimethylphenyl] -amino] -2-pyrimidinyl] -amino] -benzonitrile or a stereoisomer thereof; Or a pharmaceutically acceptable salt thereof;
(ii) tenofovir or its prodrug tenofovir disoproxyl fumarate; And
(iii) a composition comprising lamivudine. The composition of claim 2, wherein the nucleoside reverse transcriptase inhibitor is abakavir or a pharmaceutically acceptable salt thereof. 18. The method of claim 17,
(i) 4-[[4-[[4- (2-cyanoethenyl) -2,6-dimethylphenyl] -amino] -2-pyrimidinyl] -amino] -benzonitrile or a stereoisomer thereof; Or a pharmaceutically acceptable salt thereof;
(ii) tenofovir or its prodrug tenofovir disoproxyl fumarate; And
(iii) a composition comprising abakavir or a pharmaceutically acceptable salt thereof. 3. The method of claim 2,
(i) 4-[[4-[[4- (2-cyanoethenyl) -2,6-dimethylphenyl] -amino] -2-pyrimidinyl] -amino] -benzonitrile or a stereoisomer thereof; Or a pharmaceutically acceptable salt thereof;
(ii) lamivudine; And
(iii) a composition comprising emtricitabine. 3. The method of claim 2,
(i) 4-[[4-[[4- (2-cyanoethenyl) -2,6-dimethylphenyl] -amino] -2-pyrimidinyl] -amino] -benzonitrile or a stereoisomer thereof; Or a pharmaceutically acceptable salt thereof;
(ii) abakavir or a pharmaceutically acceptable salt thereof; And
(iii) a composition comprising emtricitabine. 3. The method of claim 2,
(i) 4-[[4-[[4- (2-cyanoethenyl) -2,6-dimethylphenyl] -amino] -2-pyrimidinyl] -amino] -benzonitrile or a stereoisomer thereof; Or a pharmaceutically acceptable salt thereof;
(ii) lamivudine; And
(iii) a composition comprising abakavir or a pharmaceutically acceptable salt thereof. The composition of claim 5, wherein the weight ratio of the two components in the triple combination taken on a daily basis can vary in the range of 1/10 to 10/1. The compound according to claim 1, wherein 4-[[4-[[4- (2-cyanoethenyl) -2,6-dimethylphenyl] -amino] -2-pyrimidinyl] -amino] -benzonitrile is E A composition characterized by the presence of isomers. The compound according to claim 1, wherein 4-[[4-[[4- (2-cyanoethenyl) -2,6-dimethylphenyl] -amino] -2-pyrimidinyl] -amino] -benzonitrile is (E ) -4-[[4-[[4- (2-cyanoethenyl) -2,6-dimethylphenyl] -amino] -2-pyrimidinyl] -amino] -benzonitrile hydrochloride salt Characterized in that the composition. A pharmaceutical formulation comprising the composition of any one of claims 1-24 and a pharmaceutically acceptable carrier.