WO1994014763A1 - Novel agents for inhibition of hiv infectivity and use therefor - Google Patents

Novel agents for inhibition of hiv infectivity and use therefor Download PDF

Info

Publication number
WO1994014763A1
WO1994014763A1 PCT/US1993/012276 US9312276W WO9414763A1 WO 1994014763 A1 WO1994014763 A1 WO 1994014763A1 US 9312276 W US9312276 W US 9312276W WO 9414763 A1 WO9414763 A1 WO 9414763A1
Authority
WO
WIPO (PCT)
Prior art keywords
pro
compound
alkyl
hiv
group
Prior art date
Application number
PCT/US1993/012276
Other languages
French (fr)
Inventor
Shyam K. Singh
Raymond J. Patch
Ariamala Gopalsamy
Peter V. Pallai
Original Assignee
Procept, Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Procept, Inc. filed Critical Procept, Inc.
Priority to AU58508/94A priority Critical patent/AU5850894A/en
Publication of WO1994014763A1 publication Critical patent/WO1994014763A1/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C309/00—Sulfonic acids; Halides, esters, or anhydrides thereof
    • C07C309/01—Sulfonic acids
    • C07C309/28—Sulfonic acids having sulfo groups bound to carbon atoms of six-membered aromatic rings of a carbon skeleton
    • C07C309/45—Sulfonic acids having sulfo groups bound to carbon atoms of six-membered aromatic rings of a carbon skeleton containing nitrogen atoms, not being part of nitro or nitroso groups, bound to the carbon skeleton
    • C07C309/47—Sulfonic acids having sulfo groups bound to carbon atoms of six-membered aromatic rings of a carbon skeleton containing nitrogen atoms, not being part of nitro or nitroso groups, bound to the carbon skeleton having at least one of the sulfo groups bound to a carbon atom of a six-membered aromatic ring being part of a condensed ring system
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C309/00—Sulfonic acids; Halides, esters, or anhydrides thereof
    • C07C309/01—Sulfonic acids
    • C07C309/28—Sulfonic acids having sulfo groups bound to carbon atoms of six-membered aromatic rings of a carbon skeleton
    • C07C309/45—Sulfonic acids having sulfo groups bound to carbon atoms of six-membered aromatic rings of a carbon skeleton containing nitrogen atoms, not being part of nitro or nitroso groups, bound to the carbon skeleton
    • C07C309/51—Sulfonic acids having sulfo groups bound to carbon atoms of six-membered aromatic rings of a carbon skeleton containing nitrogen atoms, not being part of nitro or nitroso groups, bound to the carbon skeleton at least one of the nitrogen atoms being part of any of the groups, X being a hetero atom, Y being any atom
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D223/00—Heterocyclic compounds containing seven-membered rings having one nitrogen atom as the only ring hetero atom
    • C07D223/14—Heterocyclic compounds containing seven-membered rings having one nitrogen atom as the only ring hetero atom condensed with carbocyclic rings or ring systems
    • C07D223/18—Dibenzazepines; Hydrogenated dibenzazepines
    • C07D223/22—Dibenz [b, f] azepines; Hydrogenated dibenz [b, f] azepines

Definitions

  • CD4 a surface glycoprotein receptor found on a subset of T lymphocytes known as CD4+ cells, is involved in Class II MHC recognition and appears to be the physio ⁇ logical receptor for Class II MHC.
  • Human CD4 is also the receptor for the gpl20 envelope glycoprotein of the human immunodeficiency virus (HIV) and is essential for virus entry into the host cell, and for membrane fusion, both of which contribute to cell-to-cell transmission of the virus and to its cytopathic effects. It is know that HIV causes AIDS by attacking the immune system and destroying CD4+ cells, thus leaving the body defenseless against attack by bacterial and other viral pathogens.
  • CD4 has been shown to be the major route of entry of HIV into CD4+ cells by binding to CD4, and then gaining entry into the cytoplasm through endocytosis.
  • a means by which to prevent HIV infection of CD4 bearing lymphocytes i.e., helper and inducer T lymphocytes
  • CD4 bearing lymphocytes i.e., helper and inducer T lymphocytes
  • helper and inducer T lymphocytes which make up approximately 60-80% of the total circulating T ly pho- cyte population
  • agents that can be used to treat AIDS-related complex (ARC) , AIDS-related dementia and non-symptomatic HIV infection ARC
  • This invention pertains to novel compounds that can abrogate HIV gpl20 binding to CD4, as demonstrated in a CD4/gpl20 binding assay. Based on these findings, the novel compounds can be used as therapeutic agents for the treatment of acquired immunodeficiency syndrome (AIDS) , as well as AIDS-related complex (ARC) , AIDS-related de ⁇ mentia and non-symptomatic HIV infection.
  • AIDS acquired immunodeficiency syndrome
  • ARC AIDS-related complex
  • AIDS-related de ⁇ mentia non-symptomatic HIV infection.
  • This invention pertains to a novel class of com ⁇ pounds that can be used to inhibit HIV infectivity.
  • compounds of this invention are repre ⁇ sented by general Formula I:
  • R 2 and R 3 are independently hydrogen, alkyl (C1-C6) , aralkyl or heteroaralkyl (wherein the aromatic portion contains C6-C10 and the alkyl portion contains C1-C3) , aryl (C6-C10) or heteroaryl (C4-C9) ; wherein the heteroatoms are selected from the group consisting of nitrogen, oxygen, sulfur and combinations thereof; wherein Ar 1 and Ar 2 are independently selected from the group consisting of phenyl, naphthyl, quinolyl, iso- quinolyl, pyridyl, thiophenyl, indolyl, benzimidazolyl, benzothiazolyl, pyrimidyl, quinazolyl, benzofuranyl and benzoxazolyl; wherein Ar 1 and Ar 2 can be substituted with one or more substituents selected from the group consisting of
  • R 2 and R 3 are defined above but exclude aromatic and heteroaromatic moieties; wherein Ar 1 and Ar 2 can be taken together, with or without their substituents, and with Y to form a fused ring system, including not limited to iminodibenzyl, carbazole, benzocarbazole, phenanthridine, phenanthrene, phenothiazine and phenoxazine.
  • novel compounds are represented by Formula II, as follows:
  • R is alkyl (C1-C4) , aryl (C6-C10) , or aral- kyl (e.g. , wherein the aromatic ring contains C6-C10 and the alkyl portion contains C1-C3) ;
  • Ar 1 and Ar 2 are independently selected from the group consisting of phenyl, naphthyl, quinolyl, isoquinolyl, pyridyl, thiophenyl, indolyl, benzimi- dazolyl, benzothiazolyl, pyrimidyl, quinazolyl, benzo ⁇ f ranyl and benzoxazolyl; wherein Ar 1 and Ar 2 can be substituted with one or more substituents selected from the group consisting of alkyl (C1-C4) , hydroxyl, alkoxyl (C1-C4) , aryloxy (C6- C10) , halo
  • Biologically active analogues of these compounds are also embraced by this application and include the addi ⁇ tion or deletion of substituents which do not detract from the biological activity of the compound, i.e., re- prise the ability of the molecule to inhibit gpl20 bind ⁇ ing.
  • Modifications to this compound include halogenat- ing, alkylating (e.g., C1-C4 alkyl groups), aralkylating, acylating, sulfonylating, as well as the addition of heteroatoms.
  • alkylating e.g., C1-C4 alkyl groups
  • aralkylating e.g., acylating, sulfonylating
  • thioureas and thiocarbamates These derivatives can be synthesized via the thioi- socyanate of 7-aminonaphthalene-l,3-disulfonic acid mono potassium salt. This synthesis is accomplished using methodology similar to that described by
  • Alkyl deriva ⁇ tives can be made by conventional routes by reacting 7-aminonaphthalene-l,3-disulfonic acid mono potassi ⁇ um salt with an activated halide (such as benzylic or allylic halides) in the presence of an aqueous base such as potassium carbonate at elevated temper ⁇ atures to yield a mixture of mono- and bis-alkylated analogues.
  • activated halide such as benzylic or allylic halides
  • mono-alkylated derivatives can be exclusively obtained under reductive amination conditions (see Scheme 3 below) .
  • the mono-alkylated derivatives can, in turn, be again alkylated to afford an unsymmetrical bis-alkylated derivative, or acylated as described above.
  • Trizenes can be made by diazotising the appropriately substituted aniline and reacting with the potassium salt of 7-amino-l , 3- naphthalenedisulfonic acid, as depicted below in Scheme 4.
  • Azo compounds are prepared by diazotization of the potassium salt of 7-amino-l,3-naphthalenedisulfonic acid and its reac ⁇ tion with the appropriately substituted aniline, as shown in Scheme 5.
  • Carbamate derivatives can be made by reacting 7-aminonaphthalene-l,3-disu- lfonic acid mono potassium salt with the correspond ⁇
  • Sulfonamides can be made by reacting 7-aminonaphthalene-l,3-disulfonic acid mono potassium salt with the corresponding a sulfonyl chloride in the presence of an aqueous base such as sodium bicarbonate, with or without the addition of a co-solvent, such as toluene or methy- lene chloride.
  • the compounds of this invention can be used to treat individuals infected with HIV, in vivo (e.g., by adminis ⁇ tration to infected individuals) . It can also be used prophyla t ically for individuals who test positive for HIV antibodies but remain asymptomatic.
  • the preparation can be used to inhibit binding of HIV to CD4 lymphocytes and to inhibit transmission of virus from an infected cell to uninfected cells by interfering with syncytia formation.
  • the preparation of this invention can be adminis- tered orally (e.g., capsular, tablet or liquid formula ⁇ tion), parenterally (e.g., intramuscularly, intravenous ⁇ ly, subcutaneously) , topically, nasally or via slow re ⁇ leasing microcarriers in dosage formulations containing physiologically acceptable vehicle and optional adjuvants and preservatives.
  • physiologically acceptable vehicles include saline, sterile water. Ringer's solu ⁇ tion, isotonic sodium chloride solutions.
  • the specific dosage level of active ingredient will depend upon a number of factors, including biological activity of the particular preparation, age, body weight, sex, general health and the clinical stage of AIDS.
  • antiviral agents which interfere with HIV viral replication can be administered in conjunction with this preparation, according to the methods of this inven- tion.
  • Co-administration of antiviral agents can effec ⁇ tively inhibit various stages of the virus life cycle, thus optimizing the therapeutic benefit of the prepara ⁇ tion of this invention, for reducing or eliminating viral infectivity and the symptoms associated therewith.
  • an HIV reverse transcriptase inhibitory agent such as zidovudine (AZT) or ddl
  • ZAT zidovudine
  • ddl can be co-administered with the compounds of this invention separately or as a single dosage that is formulated with other antiviral agent(s) .
  • the invention will be further illustrated by the following non-limiting exemplification:
  • the mobile phase consisted of 0.1% trifluoroacetic acid in water (solvent A) and 0.1% trifluoroacetic acid in acetonitrile/water (60:40) (solvent B) .
  • solvent A 0.1% trifluoroacetic acid in water
  • solvent B 0.1% trifluoroacetic acid in acetonitrile/water
  • PRO 363 Purity of PRO 363 was confirmed by analytical HPLC using a Waters 625 LC system equipped with a photodiode array detector (Waters 991) .
  • a solution of PRO 363 in water 50 ⁇ l of a 1 mg/ l solution
  • the concentration of solvent B in the mobile phase was increased to 100% at a rate of 2.7% per min.
  • PRO 363 eluted at 17.5 min relative to Orange G (Aldrich Chemical Company, lot # 04116MX) , which eluted at 8.9 min.
  • PRO 376 was carried out using the meth ⁇ odology described above for the synthesis of PRO 363 by substituting 2-benzylaniline (268.5 mg; 1.46 mmol) for 2- a inobiphenyl.
  • PRO 376 was also purified using the same system and conditions as for PRO 363. Monitoring at 400 nm, the peak eluting between 28.6 and 30.1 minutes was collected and evaporated to dryness under reduced pres ⁇ sure to afford 30 mg of pure PRO 376.
  • PRO 377 and PRO 378 were again synthesized by the methodology described for PRO 363 by using 2,2'-ethylene- dianiline diphosphate (454.6 mg; 1.46 mmol) instead of 2- a inobiphenyl.
  • the products, PRO 377 and PRO 378, were purified in a similar way as PRO 363. Again monitoring at 400 nm, the peak eluting between 19.60 and 20.62 min ⁇ utes was collected and evaporated to dryness to yield 45 mg of pure PRO 377, whereas the peak eluting between 30.0 and 32.0 minutes gave 8 mg of pure PRO 378, upon concen- tration.
  • PRO 377 and PRO 378 were confirmed as described for PRO 363.
  • PRO 377 eluted at 11.4 minutes and PRO 378 at 15.8 minutes.
  • PRO 391 was synthesized by a slight modification of the methodology described for PRO 363 where the diazonium salt suspension of 7-amino-l,3-naphthaleneidsulfonic acid was neutralized to - pH 7.5 by dropwise addition of sodi- urn hydroxide at 6-7° C. To this solution was then added a solution of iminodibenzyl (286 mg; 1.46 mmol) in metha ⁇ nol (4 mL) , dropwise with stirring at 6-7°C. The reac ⁇ tion mixture was stirred at ambient temperature over ⁇ night, filtered to remove a small amount of precipitate, and evaporated to dryness. The product was purified by preparative HPLC using the same method and conditions as for PRO 363, to give 60 mg of pure PRO 391.
  • PRO 391 Purity of PRO 391 was confirmed by analytical HPLC. Under the similar conditions as PRO 363, PRO 391 eluted at 24.5 minutes.
  • PRO 407 was accomplished using the method of PRO 391 by substituting 3-aminobiphenyl (247.9 mg; 1.46 mmol) for iminodibenzyl. The product was then purified on HPLC using the conditions of PRO 363, to afford 200 mg of PRO 407. Purity of PRO 407 was checked the same way as for PRO 363. PRO 407 eluted at 15.2 minutes.
  • PRO 376 (25 mg; 0.05 mmol) predried under vacuum at 40"C overnight) was sus ended in acetic anhydride (2mL) and refluxed under argon for 1.5 hours. The dark reac ⁇ tion mixture was evaporated to dryness under reduced pressure and the resultant solid was purified on HPLC using the method and conditions of PRO 363 to afford 15.6 mg of PRO 408.
  • PRO 414 was synthesized by the method of PRO 391 by substituting N-phenyl-l,2-phenylenediamine (269.9 mg; 1.46 mmol) for iminodibenzyl. The product was purified using the method described for PRO 363, to afford 35 mg of pure PRO 414.
  • PRO 414 Purity of PRO 414 was confirmed by analytical HPLC. Under the conditions of PRO 363, PRO 414 eluted at 15.8 minutes.
  • PRO 415 was prepared using the methodology of PRO 391 by substituting 2-phenoxyaniline (271.3 mg; 1.46 mmol) for iminodibenzyl. The product was purified by the method described for PRO 363, to give 40 mg of pure PRO 415.
  • PRO 421 was prepared using the methodology of PRO 391 by substituting a solution of 2-hydroxybiphenyl (248 mg; 1.46 mmol) in 10% sodium hydroxide (3mL) for the methanolic solution of iminodibenzyl. The product was purified by the method described for PRO 363, to give 444 mg of pure PRO 421. Confirmation of the purity of PRO 421 was again carried out by analytical HPLC. Under similar conditions as PRO 363, PRo 421 eluted at 18.5 minutes.
  • the aqueous layer was then purified directly by preparative HPLC using a Waters 600E system equipped with a PrePack® RCM cartridge column assembly (Waters Chromatography, Division of Millipore, Milford, MA) .
  • the mobile phase consisted of 0.1% trifluoroacetic acid in water (solvent A) and 0.1% trifluoroacetic acid in acetonitrile/water (60:40) (sol ⁇ vent B) .
  • solvent A trifluoroacetic acid in water
  • solvent B acetonitrile/water
  • the concentration of solvent B was increased to 100% in 40 minutes.
  • PRO 374 eluted at 16.5 minutes.
  • PRO 410 The synthesis and purification of PRO 410 was car ⁇ ried out using the methodology described above for the synthesis of PRO 374 by substituting 1-naphthylacetyl chloride with 3-phenylbenzoyl chloride (216 mg; 1 mmol) . Confirmation of purity of PRO 410 was carried out as described for PRO 374. Under similar conditions, PRO 410 eluted at 18.0 minutes.
  • the reaction mixture was stirred at room temperature for 3 hours and purified by high pressure liquid chromatogra ⁇ phy (HPLC) using a Waters 600E system equipped with Pre ⁇ Pack® RCM Cartridge column assembly (Waters Chromatogra- phy, Division of Millipore, Milford, MA) .
  • HPLC high pressure liquid chromatogra ⁇ phy
  • the mobile phase consisted of 0.1% trifluoroacetic acid in water (solvent A) and 0.1% trifluoroacetic acid in aceton- itrile/water (60:40) (solvent B) .
  • PRO 395 and PRO 420 Purity of PRO 395 and PRO 420 was confirmed by ana- lytical HPLC using a Waters 625LC system equipped with a photodiode array detector (Waters 991) .
  • a solution of PRO 395 in water 50 mL of a mg/mL solution
  • CEM cells (a human T cell leukemia line available from the American Type Culture Collection, Rockville, MD. ; 3 X 10 6 cells per ml) were suspended in RPMI 1640 (Mediatech, Washington, D.C.) with 10% fetal bovine serum
  • test compounds were dissolved in either water or dimethyl sulfoxide (DMSO) to a final concentration of 4 mg/ml.
  • DMSO dimethyl sulfoxide
  • PRO 378 50 PRO 391 50
  • PRO 415 50 PRO 421 150

Abstract

This invention pertains to compounds represented by formula (I) and physiologically acceptable salts thereof; wherein R1, X, Y, Ar1, and Ar2 are as defined in Claim 1; that can abrogate GIV gp120 binding to CD4, as demonstrated in a CD4/gp120 binding assay. Based in these findings, the compounds can be used as therapeutic agents for the treatment of acquired immunodeficiency syndrome (AIDS), as well as AIDS-related complex (ARC), AIDS-related dementia and non-symptomatic HIV infection.

Description

NOVEL AGENTS FOR INHIBITION OF HIV INFECTIVITY
AND USE THEREFOR
Background of the Invention
CD4, a surface glycoprotein receptor found on a subset of T lymphocytes known as CD4+ cells, is involved in Class II MHC recognition and appears to be the physio¬ logical receptor for Class II MHC. Human CD4 is also the receptor for the gpl20 envelope glycoprotein of the human immunodeficiency virus (HIV) and is essential for virus entry into the host cell, and for membrane fusion, both of which contribute to cell-to-cell transmission of the virus and to its cytopathic effects. It is know that HIV causes AIDS by attacking the immune system and destroying CD4+ cells, thus leaving the body defenseless against attack by bacterial and other viral pathogens. CD4 has been shown to be the major route of entry of HIV into CD4+ cells by binding to CD4, and then gaining entry into the cytoplasm through endocytosis.
Considerable effort has been expended in studying the CD4-gpl20 interaction and in trying to interfere with or inhibit that interaction, in an attempt to provide a means by which the life threatening effects of HIV infec¬ tion can be slowed or reversed. Thus far, a small number of antiviral drugs have been developed to interfere with infection of cells by HIV and its subsequent effects, such as zidovudine (also know as AZT) or ddl. A means by which to prevent HIV infection of CD4 bearing lymphocytes (i.e., helper and inducer T lymphocytes), which make up approximately 60-80% of the total circulating T ly pho- cyte population, would be of great value, particularly in light of the fact that HIV infection of such cells can cause total collapse of the immune system. It would be of further value to develop agents that can be used to treat AIDS-related complex (ARC) , AIDS-related dementia and non-symptomatic HIV infection.
Summary of Invention This invention pertains to novel compounds that can abrogate HIV gpl20 binding to CD4, as demonstrated in a CD4/gpl20 binding assay. Based on these findings, the novel compounds can be used as therapeutic agents for the treatment of acquired immunodeficiency syndrome (AIDS) , as well as AIDS-related complex (ARC) , AIDS-related de¬ mentia and non-symptomatic HIV infection.
Detailed Description of the Invention
This invention pertains to a novel class of com¬ pounds that can be used to inhibit HIV infectivity. In one embodiment, compounds of this invention are repre¬ sented by general Formula I:
Figure imgf000004_0001
and physiologically acceptable salts thereof; wherein R1 is hydrogen, C1-C6 alkyl, aralkyl (wherein the aromatic portion contains C6-C10 carbon atoms and the alkyl portion contains 1 to 3 carbon atoms) which may contain heteroatoms (e.g., oxygen, nitrogen, sulfur and combinations thereof) or R1 can be nothing if X is =N- to form an azo moiety; wherein X is S02, -N=N-, =N- or -C-(CH2)n- where n is 0, 1 or 2; wherein Y is -(CH2)n-, where n is 0, 1 or 2, -0-CH2-, -0-CH2-CH2-,
R2 R3 R2
I I I oxygen, sulfur, -CH-N-, -N- and -CH=CH-; wherein R2 and R3 are independently hydrogen, alkyl (C1-C6) , aralkyl or heteroaralkyl (wherein the aromatic portion contains C6-C10 and the alkyl portion contains C1-C3) , aryl (C6-C10) or heteroaryl (C4-C9) ; wherein the heteroatoms are selected from the group consisting of nitrogen, oxygen, sulfur and combinations thereof; wherein Ar1 and Ar2 are independently selected from the group consisting of phenyl, naphthyl, quinolyl, iso- quinolyl, pyridyl, thiophenyl, indolyl, benzimidazolyl, benzothiazolyl, pyrimidyl, quinazolyl, benzofuranyl and benzoxazolyl; wherein Ar1 and Ar2 can be substituted with one or more substituents selected from the group consisting of alkyl (C1-C4) , hydroxyl, alkoxyl (C1-C4) , aryloxy (C6- C10) , halo (e.g., fluorine, chlorine, bromine and iodine) , amino, nitro, alkylamino (C1-C4) , dialkylamino (C1-C4) , aralkylamino (wherein the aromatic ring contains C6-C10 and the alkyl portion contains C1-C3) and combi¬ nations thereof, provided that when X is =N- then Ar1 con¬ tains at least one substituent that is R2
N-R3 or OH; wherein R2 and R3 are defined above but exclude aromatic and heteroaromatic moieties; wherein Ar1 and Ar2 can be taken together, with or without their substituents, and with Y to form a fused ring system, including not limited to iminodibenzyl, carbazole, benzocarbazole, phenanthridine, phenanthrene, phenothiazine and phenoxazine.
In another embodiment of the invention, the novel compounds are represented by Formula II, as follows:
Figure imgf000006_0001
and physiologically acceptable salts thereof; wherein R2 is R3, hydrogen, alkyl (C1-C4) , aralkyl or heteroaralkyl (wherein the aromatic ring is C6-C10 and the alkyl portion is C1-C3) , acyl (C1-C6) , aracyl (C7- Cll) or heteroaracyl (C5-C10) ; wherein the heteroatoms are selected from the group consisting of oxygen, nitro¬ gen, sulfur and combinations of these; wherein R3 is -(CH2)n-Ar1-X-Ar2; wherein n is 1 or 2; wherein X is oxygen, sulfur, -CH2-, -CH2CH2-, -CH=CH-, -0-CH2- or -0-CH2CH2-; wherein Ar1 and Ar2 are independently selected from the group consisting of phenyl, naphthyl, quinolyl, iso- quinolyl, pyridyl, thiophenyl, indolyl, benzimidazolyl, benzothiazolyl, pyrimidyl, quinazolyl, benzofuranyl and benzoxazolyl; wherein Ar1 and Ar2 can be substituted with one or more substituents selected from the group consisting of alkyl (C1-C4) , hydroxyl, alkoxyl (C1-C4) , aryloxy (C6- iodine) , amino, nitro, alkylamino (C1-C4) , dialkylamino (C1-C4) , aralkylamino (wherein the aromatic ring contains C6-C10 and the alkyl portion contains C1-C3) and combi¬ nations thereof; wherein Ar1 and Ar2 can be taken together to form a fused ring system with or without their substituent and with X (up to five rings) , including but not limited to imino ibenzyl , carbazole, benzocarbazole, phenanthridine, phenanthrine, phenothiazine and phenoxazine. In yet another embodiment of the invention, novel compounds are represented by the following general Formu¬ la III:
Figure imgf000007_0001
and physiologically acceptable salts thereof;
O H S R O S
II I II I II II wherein X is -C-N-, -C-N-, -C-O- or -C-O-; wherein Y is oxygen, -CH2-, -CH2CH2-, -0-CH2-, H O
I II
-0-CH2CH2- or -N-C-; wherein R is alkyl (C1-C4) , aryl (C6-C10) , or aral- kyl (e.g. , wherein the aromatic ring contains C6-C10 and the alkyl portion contains C1-C3) ; wherein Ar1 and Ar2 are independently selected from the group consisting of phenyl, naphthyl, quinolyl, isoquinolyl, pyridyl, thiophenyl, indolyl, benzimi- dazolyl, benzothiazolyl, pyrimidyl, quinazolyl, benzo¬ f ranyl and benzoxazolyl; wherein Ar1 and Ar2 can be substituted with one or more substituents selected from the group consisting of alkyl (C1-C4) , hydroxyl, alkoxyl (C1-C4) , aryloxy (C6- C10) , halo (e.g., fluorine, chlorine, bromine and iodine) , amino, nitro, alkylamino (C1-C4) , dialkylamino (C1-C4) , aralkylamino (e.g., wherein the aromatic ring contains C6-C10 and the alkyl portion contains C1-C3) and combinations thereof; wherein Ar1 and Ar2 can be taken together with or without their substituents and with Y to form a fused ring system (up to five rings) , including but not limited to iminodibenzyl, carbazole, benzocarbazole, phenan- thridine, phenanthrine, phenothiazine and phenoxazine.
Biologically active analogues of these compounds are also embraced by this application and include the addi¬ tion or deletion of substituents which do not detract from the biological activity of the compound, i.e., re- duce the ability of the molecule to inhibit gpl20 bind¬ ing. Modifications to this compound include halogenat- ing, alkylating (e.g., C1-C4 alkyl groups), aralkylating, acylating, sulfonylating, as well as the addition of heteroatoms. Such modifications of the above structure are well within the skilled artisan.
Compounds of this invention can be synthesized using known synthetic techniques, such as those listed below:
1. Synthesis of amides: These derivatives can be synthesized by conventional routes by reacting 7- aminonaphthalene-l,3-disulfonic acid mono potassium salt with the corresponding acid chloride or acti¬ vated ester in the presence of an aqueous base such as sodium bicarbonate, with or without the addition of a co-solvent such as toluene or methylene chlo¬ ride. Akerfeldt, S., J. Med. Chem.. 14:596-600 (1971) .
Synthesis of thioureas and thiocarbamates: These derivatives can be synthesized via the thioi- socyanate of 7-aminonaphthalene-l,3-disulfonic acid mono potassium salt. This synthesis is accomplished using methodology similar to that described by
10 Braunitzer, G. et a . , Z. Physiol Chem.. 352:1730- 1732 (1971) and shown in Scheme 1. Conversion then to the corresponding thioureas and thiocarbamates can be carried out using conventional methodology.
Thiophosgene
Figure imgf000009_0001
Figure imgf000009_0002
Figure imgf000009_0003
SCHEME 1 Synthesis of alkyl derivatives: Alkyl deriva¬ tives can be made by conventional routes by reacting 7-aminonaphthalene-l,3-disulfonic acid mono potassi¬ um salt with an activated halide (such as benzylic or allylic halides) in the presence of an aqueous base such as potassium carbonate at elevated temper¬ atures to yield a mixture of mono- and bis-alkylated analogues. These derivatives are easily separated by reverse phase HPLC methods (see Scheme 2 below) ; Swiss Patent No. 235,195 Mar. 16, 1945.
Alternatively, mono-alkylated derivatives can be exclusively obtained under reductive amination conditions (see Scheme 3 below) . The mono-alkylated derivatives can, in turn, be again alkylated to afford an unsymmetrical bis-alkylated derivative, or acylated as described above.
Figure imgf000010_0001
iro-jS
Figure imgf000010_0002
Figure imgf000010_0003
SCHEME 2 RCHO
Figure imgf000011_0001
1) RCHO, Δ
Figure imgf000011_0002
2) NaBH4
SCHEME 3
Synthesis of triazeneε: Trizenes can be made by diazotising the appropriately substituted aniline and reacting with the potassium salt of 7-amino-l , 3- naphthalenedisulfonic acid, as depicted below in Scheme 4.
Figure imgf000011_0003
SCHEME 4 5. Synthesis at azo compounds: Azo compounds are prepared by diazotization of the potassium salt of 7-amino-l,3-naphthalenedisulfonic acid and its reac¬ tion with the appropriately substituted aniline, as shown in Scheme 5.
Figure imgf000012_0001
SCHEME 5
6. Synthesis of carbamates: Carbamate derivatives can be made by reacting 7-aminonaphthalene-l,3-disu- lfonic acid mono potassium salt with the correspond¬
10 ing carbamoyl chloride in the presence of an aqueous base such as sodium bicarbonate, with or without the addition of a co-solvent such as toluene or methy- lene chloride, as described above for amides.
7. Synthesis of ureas: These derivatives can be made by reacting 7-aminonaphthalene-l,3-disulfonic acid mono potassium salt with the corresponding isocyanate in dimethylformamide at elevated tempera¬ tures.
8. Synthesis of sulfonamides: Sulfonamides can be made by reacting 7-aminonaphthalene-l,3-disulfonic acid mono potassium salt with the corresponding a sulfonyl chloride in the presence of an aqueous base such as sodium bicarbonate, with or without the addition of a co-solvent, such as toluene or methy- lene chloride.
The compounds of this invention can be used to treat individuals infected with HIV, in vivo (e.g., by adminis¬ tration to infected individuals) . It can also be used prophyla t ically for individuals who test positive for HIV antibodies but remain asymptomatic. The preparation can be used to inhibit binding of HIV to CD4 lymphocytes and to inhibit transmission of virus from an infected cell to uninfected cells by interfering with syncytia formation.
The preparation of this invention can be adminis- tered orally (e.g., capsular, tablet or liquid formula¬ tion), parenterally (e.g., intramuscularly, intravenous¬ ly, subcutaneously) , topically, nasally or via slow re¬ leasing microcarriers in dosage formulations containing physiologically acceptable vehicle and optional adjuvants and preservatives. Suitable physiologically acceptable vehicles include saline, sterile water. Ringer's solu¬ tion, isotonic sodium chloride solutions. The specific dosage level of active ingredient will depend upon a number of factors, including biological activity of the particular preparation, age, body weight, sex, general health and the clinical stage of AIDS.
Other antiviral agents which interfere with HIV viral replication can be administered in conjunction with this preparation, according to the methods of this inven- tion. Co-administration of antiviral agents can effec¬ tively inhibit various stages of the virus life cycle, thus optimizing the therapeutic benefit of the prepara¬ tion of this invention, for reducing or eliminating viral infectivity and the symptoms associated therewith. For example, an HIV reverse transcriptase inhibitory agent, such as zidovudine (AZT) or ddl, can be co-administered with the compounds of this invention separately or as a single dosage that is formulated with other antiviral agent(s) . The invention will be further illustrated by the following non-limiting exemplification:
EXEMPLIFICATION
Synthesis of PRO 363:
To a solution of commercial monopotassium salt of 7- amino-l,3-naphthalenedisulfonic acid (500 mg; 1.46 mmol) in water (3 mL) and potassium carbonate (100 mg; 0.72 mmol) was added a solution of sodium nitrite (110 mg; 1.59 mmol) in water (2mL) . The resulting solution was then dropped, with stirring, into a test tube containing crushed ice (~ 10 gm) and concentrated hydrochloric acid (lmL) to precipitate out shining pink crystals of the diazonium salt. The suspension was stirred at 2°C for 30 minutes and then added dropwise to a stirred suspension of 2-aminobiophenyl (247.9 mg; 1.46 mmol) and excess potassium carbonate in water (25 L) maintained at pH >8 and - 7°C temperature. After stirring at ambient temper¬ ature for one hour, the reaction mixture was filtered and the filtrate was purified by preparative high pressure liquid chromatography (HPLC) using a Waters 600E system equipped with PrePack® RCM cartridge column assembly (Waters Chromatography, Division of Millipore, Milford, MA). The mobile phase consisted of 0.1% trifluoroacetic acid in water (solvent A) and 0.1% trifluoroacetic acid in acetonitrile/water (60:40) (solvent B) . Starting with an initial solvent mixture of 90% solvent A; 10% solvent B and a flow rate of 20 mL per minute, the stock solution (10 L) was loaded on a 40 X 300 mm DeltaPak™ C18 column (particle size = 15μM, mean pore diameter = 300A°). The concentration of solvent B was increased to 100% in 40 minutes. Monitoring at 400 nm, the peak eluting between 31.0 and 32.1 min was collected separately and evaporated under reduced pressure at 40βC to afford 54 mg of pure PRO 363.
Purity of PRO 363 was confirmed by analytical HPLC using a Waters 625 LC system equipped with a photodiode array detector (Waters 991) . A solution of PRO 363 in water (50μl of a 1 mg/ l solution) was injected onto a 3.9 X 150 mm DeltaPak™ C18 column (particle size = 5μM, mean pore diameter = 300 A") with an initial mobile phase mixture consisting of 90% solvent A and 10% solvent B and a flow rate of 1.0 mL/ in. After a run time of 2 min¬ utes, the concentration of solvent B in the mobile phase was increased to 100% at a rate of 2.7% per min. Under these conditions, PRO 363 eluted at 17.5 min relative to Orange G (Aldrich Chemical Company, lot # 04116MX) , which eluted at 8.9 min.
1H NMR (250 MHz, DMSO-d6) δ 9.28 (s, IH) , 8.28 (s, IH) , 8.12 (s, IH) , 8.04 (d, IH, JQ = 8.82 Hz) , 7.92 (dd, IH, J0 = 8.88 Hz, Jm = 1.80 Hz) 7.82 (dd, IH, J0 = 8.50 Hz, Jπ = 2.00 Hz), 7.70 (d, IH, Jm = 2.10 Hz), 7.58 - 7.40 (m, 5H) , 7.00 (d, IH, J0 = 8.56 Hz) , 4.80 ppm (br s, 2H, exch . ) .
FAB/MS 482 (M-l) .
Figure imgf000016_0001
Synthesis of PRO 376:
Synthesis of PRO 376 was carried out using the meth¬ odology described above for the synthesis of PRO 363 by substituting 2-benzylaniline (268.5 mg; 1.46 mmol) for 2- a inobiphenyl. PRO 376 was also purified using the same system and conditions as for PRO 363. Monitoring at 400 nm, the peak eluting between 28.6 and 30.1 minutes was collected and evaporated to dryness under reduced pres¬ sure to afford 30 mg of pure PRO 376.
Confirmation of purity of PRO 376 was carried out as described for PRO 363. Under similar conditions, PRO 376 eluted at 17.4 min. YL NMR (250 MHz, DMSO-d6) δ 9.22 (s, IH) , 8.26 (s, IH) , 8.10 (s, IH) , 8.00 (d, IH, JQ = 8.90 Hz), 7.86 (dd, IH, J0 = 8.94 Hz, Jm = 1.76 Hz), 7.68 (dd, IH, J0 = 8.46 Hz, Jm = 1.98 Hz), 7.60 (d, IH, Jm = 1.92 Hz), 7.36 - 7.20 (m, 5H) , 6.84 (d, IH, JQ = 8.44 Hz), 4.10 (br s, 2H, exch.) 3.82 ppm (s, 2H) .
FAB/MS 496 (M-l)
Figure imgf000017_0001
Synthesis of PRO 377 and PRO 378:
PRO 377 and PRO 378 were again synthesized by the methodology described for PRO 363 by using 2,2'-ethylene- dianiline diphosphate (454.6 mg; 1.46 mmol) instead of 2- a inobiphenyl. The products, PRO 377 and PRO 378, were purified in a similar way as PRO 363. Again monitoring at 400 nm, the peak eluting between 19.60 and 20.62 min¬ utes was collected and evaporated to dryness to yield 45 mg of pure PRO 377, whereas the peak eluting between 30.0 and 32.0 minutes gave 8 mg of pure PRO 378, upon concen- tration.
Purities of PRO 377 and PRO 378 were confirmed as described for PRO 363. PRO 377 eluted at 11.4 minutes and PRO 378 at 15.8 minutes.
1H NMR of PRO 377 (250 MHz, DMSO-d6) δ 9.38 (s, IH) , 8.30 (s, IH) , 8.14 (s, IH) , 8.11 (d, IH, Jm = 1.89 Hz),
8.08 (d, IH, J0 = 8.86 Hz), 7.95 (dd, IH, JQ = 8.92 Hz, Jm = 1.94 Hz), 7.84 (dd, IH, J0 = 8.48 Hz, Jm = 2.00 Hz), 7.21 (d, IH, J0 = 8.50 Hz), 4.00 (br s, 2H, exch.) 3.10 ppm (s, 2H) .
FAB/MS 839 (M-l) .
Figure imgf000018_0001
1H NMR of PRO 378 (250 MHz, DMSO-d6) δ 9.44 (s, IH) , 8.28 (d, IH, Jm = 1.96 Hz), 8.14 (s, IH) , 8.04 (d, IH, JQ = 8.96 Hz) , 7.82 - 7.72 (m, 2H) , 7.62 (dd, IH, J„ = 8.90 Hz, Jm = 1.90 Hz) , 7.40 - 7.28 (m, 4H) , 7.08 (d, IH, J0 = 8.50 Hz), 4.60 (br s, 2H, exch.) 2.88 - 2.62 ppm (m, 4H) .
FAB/MS 509 (M-NH2~H) .
Figure imgf000018_0002
Synthesis of PRO 391:
PRO 391 was synthesized by a slight modification of the methodology described for PRO 363 where the diazonium salt suspension of 7-amino-l,3-naphthaleneidsulfonic acid was neutralized to - pH 7.5 by dropwise addition of sodi- urn hydroxide at 6-7° C. To this solution was then added a solution of iminodibenzyl (286 mg; 1.46 mmol) in metha¬ nol (4 mL) , dropwise with stirring at 6-7°C. The reac¬ tion mixture was stirred at ambient temperature over¬ night, filtered to remove a small amount of precipitate, and evaporated to dryness. The product was purified by preparative HPLC using the same method and conditions as for PRO 363, to give 60 mg of pure PRO 391.
Purity of PRO 391 was confirmed by analytical HPLC. Under the similar conditions as PRO 363, PRO 391 eluted at 24.5 minutes.
1H NMR (250 MHZ, DMSO-d6) 6 9.31 (s, IH) , 9.02 (s, IH, exch.), 8.27 (s, 1H),,8.13 (s, IH) , 8.04 (d, IH, J0 = 8.98 Hz), 7.92 (dd, IH, J0 = 8.81 Hz, Jm = 1.88 Hz), 7.77 - 7.20 (m, 2H) , 7.17 - 7.07 (m, 4H) , 6.80 - 6.73 (m, IH) , 3.20 - 3.00 ppm (m, 4H) .
FAB/MS 509 (M-l) .
Figure imgf000019_0001
Synthesis of PRO 407:
Synthesis of PRO 407 was accomplished using the method of PRO 391 by substituting 3-aminobiphenyl (247.9 mg; 1.46 mmol) for iminodibenzyl. The product was then purified on HPLC using the conditions of PRO 363, to afford 200 mg of PRO 407. Purity of PRO 407 was checked the same way as for PRO 363. PRO 407 eluted at 15.2 minutes.
1H NMR (250 MHz, DMSO-d6) δ 9.36 (d, IH Jm = 1.65 Hz) , 8.26 (d, IH, Jm = 1.48 Hz), 8.10 (s, IH) , 8.00 - 7.90 (m, 2H) , 7.52 - 7.41 (m, 6H) , 7.03 - 6.98 (m, 2H) , 4.65 ppm (br s, 2H, exch. ) .
FAB/MS 482 (M-l) .
Figure imgf000020_0001
Synthesis of PRO 408:
PRO 376 (25 mg; 0.05 mmol) predried under vacuum at 40"C overnight) was sus ended in acetic anhydride (2mL) and refluxed under argon for 1.5 hours. The dark reac¬ tion mixture was evaporated to dryness under reduced pressure and the resultant solid was purified on HPLC using the method and conditions of PRO 363 to afford 15.6 mg of PRO 408.
Confirmation of the purity of PRO 408 was carried out by the method of PRO 363. PRO 408 eluted at 17.0 minutes.
1H NMR (250 MHz, DMSO-d6) δ 9.56 (s, IH, exch.), 9.40 (d, IH, Jm = 1.76 Hz), 8.28 (s, IH) , 8.14 (s, IH) , 8.06 (d, IH, J0 = 8.98 Hz), 7.94 (s, IH) , 7.91 (dd, IH, JQ = 8.84 Hz, Jm = 1.86 Hz) , 7.85 - 7.73 (m, 3H) , 7.33 - 7.16 (m, 4H) , 4.16 (s, 2H) , 2.02 ppm (s, 3H) .
FAB/MS 538 (M-l) .
Figure imgf000021_0001
Synthesis of PRO 414:
PRO 414 was synthesized by the method of PRO 391 by substituting N-phenyl-l,2-phenylenediamine (269.9 mg; 1.46 mmol) for iminodibenzyl. The product was purified using the method described for PRO 363, to afford 35 mg of pure PRO 414.
Purity of PRO 414 was confirmed by analytical HPLC. Under the conditions of PRO 363, PRO 414 eluted at 15.8 minutes.
1H NMR (250 MHz, DMSO-d6) δ 9.20 (s, IH) , 8.24 (s, IH) , 8.10 (s, IH) , 8.01 (d, IH, J0 = 9.00 Hz) , 7.88 (dd, IH, J0 = 8.82 Hz, Jm = 1.87 Hz) , 7.74 (d, IH, Jm = 2.15 Hz), 7.59 (dd, IH, J0 = 8.53 Hz, Jm = 2.20 Hz) , 7.27 - 7.20 (m, 2H) , 6.99 - 6.77 (m, 4H) , 4.50 ppm (br s, 3H, exch.) .
FAB/MS 497 (M-l) .
Figure imgf000021_0002
Synthesis of PRO 415:
PRO 415 was prepared using the methodology of PRO 391 by substituting 2-phenoxyaniline (271.3 mg; 1.46 mmol) for iminodibenzyl. The product was purified by the method described for PRO 363, to give 40 mg of pure PRO 415.
Confirmation of the purity of PRO 415 was again carried out by analytical HPLC. Under similar conditions as PRO 363, PRO 415 eluted at 18 minutes. 1H NMR (250 MHz, DMSO-d6) δ 9.16 (s, IH) , 8.23 (s, IH) , 8.09 (s, IH) , 8.00 (d, IH, J0 = 8.98 Hz) , 7.94 (s, IH) , 7.84 (dd, IH, J0 = 8.91 Hz, Jm = 1.76 Hz), 7.67 (dd, IH, J0 = 8.50 Hz, Jm = 2.07 Hz), 7.46 - 7.35 (m, 2H) , 7.32 (d, IH, Jm = 2.06 Hz), 7.20 - 7.08 (m, 2H) , 7.00 (d, IH, J„ = 8.35 Hz), 5.10 ppm (br s, 2H, exch.). FAB/MS 497 (M-l) .
Figure imgf000022_0001
Synthesis of PRO 421:
PRO 421 was prepared using the methodology of PRO 391 by substituting a solution of 2-hydroxybiphenyl (248 mg; 1.46 mmol) in 10% sodium hydroxide (3mL) for the methanolic solution of iminodibenzyl. The product was purified by the method described for PRO 363, to give 444 mg of pure PRO 421. Confirmation of the purity of PRO 421 was again carried out by analytical HPLC. Under similar conditions as PRO 363, PRo 421 eluted at 18.5 minutes. NMR (250 MHz, DMSO-d6) δ 9.36 (s, IH) , 8.28 (s, IH) , 8.15 (ε, IH) , 8.07 (d, IH, JQ = 8.66 Hz) , 7.94 (dd, IH, J0 = 8.60 Hz, Jm = 1.86 Hz), 7.86 (m, 2H) , 7.65 (m, 2H) , 7.32 - 7.48 (m, 3H) , 7.16 (d, IH, J0 = 8.60 Hz), 4.64 ppm (br s, 2H, exch.).
FAB/MS 483 (M-l) .
Figure imgf000023_0001
Synthesis of PRO 374:
To a solution of 7-amino-l,3-naphthalenedisulfonic acid, monopotassium salt (341 mg; 1 mmol) and potassium bicarbonate (300 mg; 3 mmol) in water (5 mL) was added a solution of 1-naphthylacetyl chloride (302 mg; 1.5 mmol) in toluene (5 L) and the resulting bi-phasic mixture was stirred at room temperature for 12h. The toluene layer was removed and the aqueous layer was acidified with IN HC1 and extracted with ether (3X5 mL) . The aqueous layer was then purified directly by preparative HPLC using a Waters 600E system equipped with a PrePack® RCM cartridge column assembly (Waters Chromatography, Division of Millipore, Milford, MA) . The mobile phase consisted of 0.1% trifluoroacetic acid in water (solvent A) and 0.1% trifluoroacetic acid in acetonitrile/water (60:40) (sol¬ vent B) . Starting with an initial solvent mixture of 90% solvent A; 10% solvent B and a flow rate of 20 L per minute, the stock solution (10 L) was loaded on a 40 X 300 mm DeltaPak™ C18 column (particle size = 15μM, mean pore diameter = 300A"). The concentration of solvent B was increased to 100% in 40 minutes. Monitoring at 280 nm, the major peak was collected and concentrated to afford 280 mg of pure PRO 374. Purity of PRO 374 was confirmed by analytical HPLC using a Waters 605 LC system according to the conditions set forth for PRO 363. Under these conditions, PRO 374 eluted at 16.5 minutes.
1H NMR (250 MHz, DMSO-d6) δ 10.72 (s, IH, exch.), 8.71 (s, IH) , 8.3 - 8.1 (m, 3H) , 8.02 (s, IH) , 8.0 - 7.7 (m, 3H) , 7.7 - 7.4 (m, 4H) , 4.22 (s, 2H) .
Figure imgf000024_0001
Synthesis of PRO 410:
The synthesis and purification of PRO 410 was car¬ ried out using the methodology described above for the synthesis of PRO 374 by substituting 1-naphthylacetyl chloride with 3-phenylbenzoyl chloride (216 mg; 1 mmol) . Confirmation of purity of PRO 410 was carried out as described for PRO 374. Under similar conditions, PRO 410 eluted at 18.0 minutes.
1H NMR (250 MHz, DMSO-d6) δ 10.77 (s, IH, exch.), 8.91 (s, IH) , 8.32 (s, IH) , 8.22 (s, IH) , 8.1 - 7.7 (m, 7H) , 7.7 - 7.6 (m, IH) , 7.6 - 7.5 (m, IH) , 7.5 - 7.4 (m, IH) .
Figure imgf000025_0001
Synthesis of PRO 395 and PRO 420:
To a suspension of commercial monopotassium salt of 7-amino-l,3-disulfonic acid (341 mg; 1 mmol) in DMSO (5 mL) was added n-butyl lithium (1.2 L of 2.5M in hexanes; 3 mmol) dropwise at room temperature. The bright orange mixture was stirred at room temperature for 15 minutes. The reaction mixture was cooled to 15°C and 2- bromomethylbiphenyl (741 mg; 3 mmol) was added dropwise. The reaction mixture was stirred at room temperature for 3 hours and purified by high pressure liquid chromatogra¬ phy (HPLC) using a Waters 600E system equipped with Pre¬ Pack® RCM Cartridge column assembly (Waters Chromatogra- phy, Division of Millipore, Milford, MA) . The mobile phase consisted of 0.1% trifluoroacetic acid in water (solvent A) and 0.1% trifluoroacetic acid in aceton- itrile/water (60:40) (solvent B) . Starting with an ini¬ tial solvent mixture of 90% solvent A; 10% solvent B and a flow rate of 20 mL per minute, the stock solution (10 mL) was loaded on 40 X 300 mm DeltaPak™ C18 column (parti¬ cle size = 15 mM, mean pore diameter = 300 A"). The concentration of solvent B was increased to 100% in 25 minutes. Monitoring at 220 nm, the peaks at 20.13 min¬ utes and 35.78 minutes were collected separately and evaporated under reduced pressure at 25"C to afford PRO 395 (92 mg) and PRO 420 (132 mg) , respectively.
Purity of PRO 395 and PRO 420 was confirmed by ana- lytical HPLC using a Waters 625LC system equipped with a photodiode array detector (Waters 991) . A solution of PRO 395 in water (50 mL of a mg/mL solution) was injected onto a 3.9 X 150 mm DeltaPak™ C18 column (particle size = 5 mM, mean pore diameter = 300 A") with an initial mobile phase mixture consisting of 90% solvent A and 10% solvent B and a flow rate of lmL/min. After a run time of 2 minutes the concentration of solvent B was increased to 100% at a rate of 2.7% per minute. Under these condi¬ tions PRO 395 eluted at 17.6 minute and PRO 420 eluted at 29.1 minute relative to Orange G (Aldrich Chemical Compa¬ ny, lot #04116 MX), which eluted at 8.9 min.
1H NMR for PRO 395 (250 MHz, DMSO-d6) δ 8.15 (s, IH) , 8.05 (s, IH) , 7.9 (s, IH) , 7.7 (d, J = 8.9 Hz, IH) , 7.6 (m, IH) , 7.5 - 7.2 (m, 8H) , 6.95 (dd, J, = 8.9 Hz, J2 = 1.5 Hz, IH) , 4.25 (m, 2H) . FAB/MS 468 (M-l) .
Figure imgf000026_0001
1H NMR for PRO 420 (250 MHz, DMSO-d6) δ 8.1 (S, IH) , 7.85 (s, IH) , 7.65 (d, J = 8.9 Hz, IH) , 7.6 - 7.05 (m, 19H) , 6.85 (d, J = 8.9 Hz, IH) , 4.5 (s, 4H) .
FAB/MS 634 (M-l) .
Figure imgf000027_0001
5 CD4/σpl20 Binding Assay
CEM cells (a human T cell leukemia line available from the American Type Culture Collection, Rockville, MD. ; 3 X 106 cells per ml) were suspended in RPMI 1640 (Mediatech, Washington, D.C.) with 10% fetal bovine serum
10 (JRH Biosciences, Lenexa, KS) plus 0.1% sodium azide. lOOμl of the suspension were added to each tube. Gener¬ ally, the test compounds were dissolved in either water or dimethyl sulfoxide (DMSO) to a final concentration of 4 mg/ml. Various dilutions (1:40, 1:200, 1:200) of test
15 compound were added to the tubes and incubated for 2 hr at 25CC. Next, gpl20 (American BiO-Technologies, Inc., Cambridge, MA) , diluted in RPMI buffer, was added to a final concentration of 10 nM. The solution was then incubated overnight (~ 16 hours) at 37°C.
20 Cells were washed thoroughly with phosphate-buffered , saline containing 10% fetal bovine serum and 0.1% sodium azide. To reveal bound gpl20, monoclonal antibody spe¬ cific for gpl20 (NEN-Dupont, NEA-9284) was then incubated with the cells at a concentration of lμg/ml (lOOμl per tube) for 30 minutes on ice. The cells were washed thor¬ oughly as before and stained with goat antimouse immuno- globulin (Boehringer Mannhein Biochemicals, Indianapolis, IN) which was labeled with fluorescein (50μl per tube) for 30 minutes on ice. The washed cells were analyzed for fluorescence on a FACScan™ (Becton Dickinson) .
The results of screening the test compounds in the CD4/gpl20 assay with CEM cells are shown in Table 1.
Table 1: Inhibition of CD4-gpl20 Binding
Compound Icso (μg/mL.
PRO 374 250
PRO 410 250
PRO 395 225 PRO 420 75
PRO 363 175
PRO 376 175
PRO 377 150
PRO 378 50 PRO 391 50
PRO 407 150
PRO 408 200
PRO 414 50
PRO 415 50 PRO 421 150
Eguivalents
Those skilled in the art will recognize, or be able to ascertain using not more than routine experimentation, many equivalents to the specific embodiments of the in- vention described herein. Such equivalents are intended to be encompassed by the following claims.

Claims

CLAIMS 1. A compound represented by the formula:
Figure imgf000029_0001
and physiologically acceptable salts thereof; wherein R1 is hydrogen, alkyl, aralkyl which may contain heteroatoms or R1 can be nothing if X is =N- to form an azo moiety;
O
II wherein X is S02, -N=N- , =N- or -C- (CH2) n- where n is O, 1 or 2 ; wherein Y is ~ (CH2) n- , where n is 0 , 1 or 2 , -O-
CH2- , -0-CH2-CH2-,
R2 R3 R2
I I I oxygen, sulfur, -CH-N-, -N- and -CH=CH-; wherein R2 and R3 are independently hydrogen, alkyl, aralkyl or heteroaralkyl, aryl or heteroaryl; wherein the heteroatoms are selected from the group consisting of nitrogen, oxygen, sulfur and combina¬ tions thereof; wherein Ar1 and Ar2 are independently selected from the group consisting of phenyl, naphthyl, quin- olyl, isoquinolyl, pyridyl, thiophenyl, indolyl, benzimidazolyl, benzothiazolyl, pyrimidyl, quinazo- lyl, benzofuranyl and benzoxazolyl; wherein Ar1 and Ar2 can be substituted with one or more substituents selected from the group con¬ sisting of alkyl, hydroxyl, alkoxyl, aryloxy, halo, amino, nitro, alkylamino, dialkylamino, aralkylamino and combinations thereof, provided that when X is
=N- then Ar1 contains at least one substituent that is R2
Figure imgf000030_0001
or OH; wherein R2 and R3 are defined above but ex¬ clude aromatic and heteroaromatic moieties; and wherein Ar1 and Ar2 can be taken together, with or without their substituents, and with Y to form a fused ring system.
2. A compound of Claim 1 wherein AR1 and AR2 taken to¬ gether form iminodibenzyl, carbazole, benzocar- bazole, phenanthridine, phenanthrene, phenothiazine and phenoxazine.
3. A compound represented by the forumla:
Figure imgf000030_0002
and physiologically acceptable salts thereof; wherein R2 is R3, hydrogen, alkyl, aralkyl or heteroaralkyl, acyl, aracyl or heteroaracyl; wherein the heteroatoms are selected from the group consist- ing of oxygen, nitrogen, sulfur and combinations of these; wherein R3 is -(CH2)n-Ar1-X-Ar2; wherein n is 1 or 2 ; wherein X is oxygen, sulfur, -CH2~, -CH2CH2~, - CH=CH-, -0-CH2- or -0-CH2CH2-; wherein Ar1 and Ar2 are independently selected from the group consisting of phenyl, naphthyl, quin- olyl, isoquinolyl, pyridyl, thiophenyl, indolyl, benzimidazolyl, benzothiazolyl, pyrimidyl, quina- zolyl, benzofuranyl and benzoxazolyl; wherein Ar1 and Ar2 can be substituted with one or more substituents selected from the group con¬ sisting of alkyl, hydroxyl, alkoxyl, aryloxy, halo, amino, nitro, alkylamino, dialkylamino, aralkylamino and combinations thereof; wherein Ar1 and Ar2 can be taken together to form a fused ring system with or without their substituent and with X.
4. A compound of Claim 3 wherein AR1 and AR2 taken to¬ gether form iminodibenzyl, carbazole, benzocar¬ bazole, phenanthridine, phenanthrene, phenothiazine and phenoxazine.
5. A compound represented by the formula:
Figure imgf000031_0001
and physiologically acceptable salts thereof; O H S R O S
II I II I II II wherein X is -C-N-, -C-N-, -C-O- or -C-O-; wherein Y is oxygen, -CH2-, -CH2CH2~, -0-CH2~, H O
I II
-0-CH2CH2- or -N-C-; wherein R is alkyl, aryl or aralkyl; wherein Ar1 and Ar2 are independently selected from the group consisting of phenyl, naphthyl, quin- olyl, isoquinolyl, pyridyl, thiophenyl, indolyl, benzimidazolyl, benzothiazolyl, pyrimidyl, quina- zolyl, benzofuranyl and benzoxazolyl; wherein Ar1 and Ar2 can be substituted with one or more substituents selected from the group con¬ sisting of alkyl, hydroxyl, alkoxyl, aryloxy, halo, amino, nitro, alkylamino, dialkylamino, aralkylamino and combinations thereof; and wherein Ar1 and Ar2 can be taken together with or without their substituents and with Y to form a fused ring system.
6. A compound of Claim 5 wherein AR1 and AR2 taken to¬ gether form iminodibenzyl, carbazole, benzocar¬ bazole, phenanthridine, phenanthrene, phenothiazine and phenoxazine.
7. A composition for inhibiting CD4/gpl20 binding com¬ prising an inhibitory amount of the compound of Claim 1, 3 or 5, in a physiologically acceptable vehicle.
8. A compound of Claim 1, 3 or 5, for use in therapy for inhibiting CD4/gpl20 binding in an HIV infected individual.
9. A compound of Claim 1, 3 or 5, for use in therapy for inhibiting HIV infectivity of CD4+ cells in an
HIV infected individual.
10. A compound of Claim 1, 3 or 5, for use in therapy for inhibiting HIV-induced syncytia formation in an HIV infected individual.
11. Use of the compound of Claim 1, 3 or 5, for the manufacture of a medicament for use inhibiting CD4/gpl20 binding in an HIV infected individual.
12. Use of the compound of Claim 1, 3 or 5, for the manufacture of a medicament for use in inhibiting HIV infectivity of CD4+ cells in an HIV infected individual.
13. Use of the compound of Claim 1, 3 or 5, for the manufacture of a medicament for use in inhibiting HIV-induced syncytia formation in an HIV infected individual.
PCT/US1993/012276 1992-12-23 1993-12-16 Novel agents for inhibition of hiv infectivity and use therefor WO1994014763A1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU58508/94A AU5850894A (en) 1992-12-23 1993-12-16 Novel agents for inhibition of hiv infectivity and use therefor

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US99676892A 1992-12-23 1992-12-23
US07/996,768 1992-12-23

Publications (1)

Publication Number Publication Date
WO1994014763A1 true WO1994014763A1 (en) 1994-07-07

Family

ID=25543291

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1993/012276 WO1994014763A1 (en) 1992-12-23 1993-12-16 Novel agents for inhibition of hiv infectivity and use therefor

Country Status (2)

Country Link
AU (1) AU5850894A (en)
WO (1) WO1994014763A1 (en)

Cited By (18)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1995014479A1 (en) * 1993-11-23 1995-06-01 Procept, Inc. Condensation polymer of an aromatic sulfonic acid and an aldehyde for inhibiting hiv infectivity
WO1996025399A1 (en) * 1995-02-17 1996-08-22 The United States Of America, Represented By The Secretary, Department Of Health And Human Services Novel aroylaniline derivatives, compositions and uses thereof in the treatment of viral infections
US5612341A (en) * 1995-06-07 1997-03-18 Biotech Research Laboratories Brominated hexahydroxybiphenyl derivatives
US7973069B2 (en) 2004-07-14 2011-07-05 Ptc Therapeutics, Inc. Methods for treating hepatitis C
US7989462B2 (en) 2003-07-03 2011-08-02 Myrexis, Inc. 4-arylamin-or-4-heteroarylamino-quinazolines and analogs as activators of caspases and inducers of apoptosis and the use thereof
US8053440B2 (en) 2007-02-01 2011-11-08 Resverlogix Corporation Compounds for the prevention and treatment of cardiovascular diseases
US8114995B2 (en) 2008-06-26 2012-02-14 Resverlogix Corp. Methods of preparing quinazolinone derivatives
US8258145B2 (en) 2005-01-03 2012-09-04 Myrexis, Inc. Method of treating brain cancer
US8309562B2 (en) 2003-07-03 2012-11-13 Myrexis, Inc. Compounds and therapeutical use thereof
US8410109B2 (en) 2005-07-29 2013-04-02 Resverlogix Corp. Pharmaceutical compositions for the prevention and treatment of complex diseases and their delivery by insertable medical devices
US8952021B2 (en) 2009-01-08 2015-02-10 Resverlogix Corp. Compounds for the prevention and treatment of cardiovascular disease
US9073878B2 (en) 2012-11-21 2015-07-07 Zenith Epigenetics Corp. Cyclic amines as bromodomain inhibitors
US9238640B2 (en) 2009-03-18 2016-01-19 Resverlogix Corp. Anti-inflammatory agents
US9271978B2 (en) 2012-12-21 2016-03-01 Zenith Epigenetics Corp. Heterocyclic compounds as bromodomain inhibitors
US9610251B2 (en) 2011-11-01 2017-04-04 Resverlogix Corp. Pharmaceutical compositions for substituted quinazolinones
US9757368B2 (en) 2009-04-22 2017-09-12 Resverlogix Corp. Anti-inflammatory agents
US9765039B2 (en) 2012-11-21 2017-09-19 Zenith Epigenetics Ltd. Biaryl derivatives as bromodomain inhibitors
US10111885B2 (en) 2015-03-13 2018-10-30 Resverlogix Corp. Compositions and therapeutic methods for the treatment of complement-associated diseases

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
CHEMICAL ABSTRACTS, vol. 94, no. 6, 9 February 1981, Columbus, Ohio, US; abstract no. 32161k, page 79; *
G.T. TAN, ET AL.:: "Potential anti-AIDS naphthalenesulphonic acid derivatives. Synthesis and inhibition of HIV-1 induced cytopathogenesis adn HIV-1 and Hiv-2 reverse transcriptase activities", JOURNAL OF MEDICINAL CHEMISTRY,, vol. 35, no. 26, 25 December 1992 (1992-12-25), WASHINGTON, DC, US,, pages 4846 - 4853 *
P. MOHAN, ET AL.:: "Anti-HIV-1 and HIV-2 activity of naphthalenedisulphonic acid derivatives. Inhibition of cytopatho- genesis, giant cell formation, and reverse transcriptase activity", BIOCHEMICAL PHARMACOLOGY,, vol. 41, no. 4, 15 February 1991 (1991-02-15), pages 642 - 646 *
P. MOHAN, ET AL.:: "Inhibition of HIV replication by naphthalenemonosulphonic acid derivatives and a bisnaphthalene- disulphonic acid compound", LIFE SCIENCES,, vol. 47, no. 12, 1990, pages 993 - 999 *
P. MOHAN, ET AL.:: "Potential anti-AIDS agents. Synthesis and antiviral activity of naphthalenesulphonic acid derivatives gainst HIV-1 and HIV-2", JOURNAL OF MEDICINAL CHEMISTRY,, vol. 34, no. 1, January 1991 (1991-01-01), pages 212 - 217 *

Cited By (32)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5614559A (en) * 1993-11-23 1997-03-25 Procept Inc. Compound for inhibiting HIV infectivity
US5677343A (en) * 1993-11-23 1997-10-14 Procept, Inc. Compound for inhibiting HIV infectivity
US6075050A (en) * 1993-11-23 2000-06-13 Procept, Inc. Compound for inhibiting HIV infectivity
US6365625B1 (en) 1993-11-23 2002-04-02 Procept, Inc. Compound for inhibiting HIV infectivity
WO1995014479A1 (en) * 1993-11-23 1995-06-01 Procept, Inc. Condensation polymer of an aromatic sulfonic acid and an aldehyde for inhibiting hiv infectivity
WO1996025399A1 (en) * 1995-02-17 1996-08-22 The United States Of America, Represented By The Secretary, Department Of Health And Human Services Novel aroylaniline derivatives, compositions and uses thereof in the treatment of viral infections
US5681832A (en) * 1995-02-17 1997-10-28 The United States Of America As Represented By The Department Of Health And Human Services Aroylaniline compounds, pharmaceutical compositions, and methods of using same to inhibit viral activity
US5612341A (en) * 1995-06-07 1997-03-18 Biotech Research Laboratories Brominated hexahydroxybiphenyl derivatives
US8309562B2 (en) 2003-07-03 2012-11-13 Myrexis, Inc. Compounds and therapeutical use thereof
US7989462B2 (en) 2003-07-03 2011-08-02 Myrexis, Inc. 4-arylamin-or-4-heteroarylamino-quinazolines and analogs as activators of caspases and inducers of apoptosis and the use thereof
US7973069B2 (en) 2004-07-14 2011-07-05 Ptc Therapeutics, Inc. Methods for treating hepatitis C
US8258145B2 (en) 2005-01-03 2012-09-04 Myrexis, Inc. Method of treating brain cancer
US8410109B2 (en) 2005-07-29 2013-04-02 Resverlogix Corp. Pharmaceutical compositions for the prevention and treatment of complex diseases and their delivery by insertable medical devices
US8053440B2 (en) 2007-02-01 2011-11-08 Resverlogix Corporation Compounds for the prevention and treatment of cardiovascular diseases
US8889698B2 (en) 2007-02-01 2014-11-18 Resverlogix Corp. Compounds for the prevention and treatment of cardiovascular diseases
US9199990B2 (en) 2007-02-01 2015-12-01 Resverlogix Corp. Compounds for the prevention and treatment of cardiovascular diseases
US10532054B2 (en) 2007-02-01 2020-01-14 Resverlogix Corp. Compounds for the prevention and treatment of cardiovascular diseases
US8114995B2 (en) 2008-06-26 2012-02-14 Resverlogix Corp. Methods of preparing quinazolinone derivatives
US8952021B2 (en) 2009-01-08 2015-02-10 Resverlogix Corp. Compounds for the prevention and treatment of cardiovascular disease
US9238640B2 (en) 2009-03-18 2016-01-19 Resverlogix Corp. Anti-inflammatory agents
US11407719B2 (en) 2009-03-18 2022-08-09 Resverlogix Corp. Anti-inflammatory agents
US10882828B2 (en) 2009-03-18 2021-01-05 Resverlogix Corp. Anti-inflammatory agents
US10131640B2 (en) 2009-03-18 2018-11-20 Resverlogix Corp. Anti-inflammatory agents
US9757368B2 (en) 2009-04-22 2017-09-12 Resverlogix Corp. Anti-inflammatory agents
US10016426B2 (en) 2011-11-01 2018-07-10 Resverlogix Corp. Pharmaceutical compositions for substituted quinazolinones
US9610251B2 (en) 2011-11-01 2017-04-04 Resverlogix Corp. Pharmaceutical compositions for substituted quinazolinones
US9765039B2 (en) 2012-11-21 2017-09-19 Zenith Epigenetics Ltd. Biaryl derivatives as bromodomain inhibitors
US9073878B2 (en) 2012-11-21 2015-07-07 Zenith Epigenetics Corp. Cyclic amines as bromodomain inhibitors
US9278940B2 (en) 2012-11-21 2016-03-08 Zenith Epigenetics Corp. Cyclic amines as bromodomain inhibitors
US9271978B2 (en) 2012-12-21 2016-03-01 Zenith Epigenetics Corp. Heterocyclic compounds as bromodomain inhibitors
US10111885B2 (en) 2015-03-13 2018-10-30 Resverlogix Corp. Compositions and therapeutic methods for the treatment of complement-associated diseases
US10772894B2 (en) 2015-03-13 2020-09-15 Resverlogix Corp. Compositions and therapeutic methods for the treatment of complement-associated diseases

Also Published As

Publication number Publication date
AU5850894A (en) 1994-07-19

Similar Documents

Publication Publication Date Title
WO1994014763A1 (en) Novel agents for inhibition of hiv infectivity and use therefor
CA2178272C (en) Antiviral monocyclic polyamines
US5583131A (en) Aromatic-linked polyamine macrocyclic compounds with anti-HIV activity
ES2226996T3 (en) NEW DERIVATIVES OF CARBOXYL ACIDS, THEIR OBTAINING AND EMPLOYMENT.
JPH09504535A (en) Novel benzoylguanidine derivatives, their preparation and use in pharmaceutical compositions
BG98616A (en) 1,3-oxathiolane nucleoside analoques
JP2988944B2 (en) Arylalkoxycoumarins, their production and therapeutic agents containing them
IE60332B1 (en) N-(2'aminophenyl)-benzamide-derivatives, process for their preparation and their use in the control of neoplastic diseases
KR20050040906A (en) Processes for the preparation of 4-[[4-[[4-(2-cyanoethenyl)-2,6-dimethylphenyl]amino]-2-pyrimidinyl]amino]benzonitrile
EP0683164B1 (en) Tocopherol derivatives
NO309567B1 (en) Process for the preparation of 1,1 '- [1,4-phenylene bis
CA2073633C (en) Use of xanthine derivatives for the treatment of secondary nerve cell damage and functional disorders after cranio-cerebral traumas
CN101463014A (en) Diaryl benzo pyridine derivative, and preparation and use thereof
EP0361831B1 (en) Antiviral nucleoside combination
JPH11500130A (en) Antiviral triaza compounds
WO1992016494A1 (en) Long chain antiviral compounds
DK165985B (en) 1-CYTOSIN DERIVATIVES, PROCEDURES FOR THEIR PREPARATION, AND THE USE OF THEM IN AND FOR THE PREPARATION OF MEDICINAL PRODUCTS
WO1988004293A1 (en) Antifolate agents
EP0113910B1 (en) Isocarbostyril derivatives, processes for their preparation and pharmaceutical compositions containing them
Bycroft et al. Synthesis of capreomycidine and epicapreomycidine, the epimers of α-(2-iminohexahydropyrimid-4-yl) glycine
US5574040A (en) Pyrimidine compounds and methods of use to derivatize neighboring lysine residues in proteins under physiologic conditions
KR0140021B1 (en) Pyrimidine acyclonucleoside derivatives
CA2083798A1 (en) 5,6-dihydro-dibenz(b,e) azepine-6,11-dione-11-oximes
NL8202227A (en) 4-AMINO.BENZYLAMINE DERIVATIVES AND METHODS FOR THEIR PREPARATION AND USE AS PHARMACEUTICS.
CN108640893B (en) Dipeptidyl peptidase-IV inhibitor compound and preparation method and application thereof

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AT AU BB BG BR BY CA CH CZ DE DK ES FI GB HU JP KP KR KZ LK LU MG MN MW NL NO NZ PL PT RO RU SD SE SK UA VN

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FR GB GR IE IT LU MC NL PT SE BF BJ CF CG CI CM GA GN ML MR NE SN TD TG

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

Ref country code: DE

Ref legal event code: 8642

122 Ep: pct application non-entry in european phase
NENP Non-entry into the national phase

Ref country code: CA