AU710536B2 - Triazine containing compounds useful as antiviral agents - Google Patents

Description

Triazine Containing Compounds Useful as Antiviral Agents This invention relates to new triazine ring containing compounds which are useful in treating viral infections and in particular human respiratory syncytial virus [HRSV]. This invention also relates to methods of treating viral infections and pharmaceutical compositions therefor and also the compounds for use as pharmaceuticals.

Background of the Invention Human respiratory syncytial virus[HRSV] was first discovered in 1956 and is worldwide in distribution. It is an important cause of upper and lower respiratory tract disease causing illness in infants and young children resulting in approximately 100 000 hospitalizations and 5000 deaths yearly in the United States (Chanock, R. Kim, H. Brandt, C. and Parrott, R. H. 1982. Respiratory syncytial virus, pp 471-489, in Viral Infections of Humans, Second Edition, A.S. Evans, editor (Plenum Press, NY). Glezen, Taber, L. Frank, and Kasel, J. 1986. Risk of primary infection and reinfection with respiratory syncytial virus. Am. J. Dis. Chil. 140;543-546. MacDonald, Hall, C. Suffin, Alexson, Harris, P. and Manning J. A. 1982. Respiratory syncytial virus infection in infants with congenital heaA disease. New England Journal of Medicine 307;397-400.

About 30% of hospitalized young children with acute respiratory disease have respiratory syncytial virus infection. In older children and adults the disease is milder. HRSV appears to also be a major cause (equivalent to influenza) of morbidity and mortality in the elderly. (Fleming, D. M. and Cross, K. W. 1993. Respiratory syncytial virus or influenza? Lancet 342;1507-1510.). Infections with respiratory syncytial virus are referable to all segments of the respiratory tract, are usually associated with fever, cough, runny nose, and fatigue, and are diagnosed clinically as bronchitis, bronchiolitis, pneumonia, croup, or viral infection. In older children and adults the virus is generally limited to replication in the upper respiratory tract. Infants may be more severely involved when the virus extends into the lungs. Lung damage can be permanent.

Primary infection with respiratory syncytial virus occurs early in life, usually before 4 years of age. Among children, illness caused by this virus tends to occur at least once each year in rather sharply defined outbreaks of several months duration. Epidemics are sharply circumscribed, generally for 3 to 5 months. In family studies, children in early school years frequently introduce the virus into the home, infecting younger members of the family more severely than other family members. The S.1 30 clinical consequence of infection is most severe on first experience and becomes milder in older individuals who are immunologically experienced.

.The effects of respiratory syncytial virus can range from unapparent infection to severe pneumonia and death. Inflammation of the respiratory track is responsible for most symptoms.

Complete recovery in most cases occurs in one to three weeks with the production of antibody which 35 appears to persist throughout life. In the United States about 30 percent of one year old infants and percent of five year old children have circulating respiratory syncytial virus antibody. Reinfection in older infants, children, and adults with antibody gives nostly mild upper respiratory illnesses in the form of colds. In infants and young children, the infection is treated with ribavirin, a broad-spectrum Libc/O2390 antiviral. Use of this compound is severely limited by toxicity. There is therefore a great need for a new therapeutic agent for the treatment of HRSV infection.

US. 5 359 131 teaches sulfonic acid stilbenes which block the infection of cells by herpes simplex virus (HSV), human immunodeficiency virus (HIV) and cytomegalovirus

(CMV).

The present invention relates to novel triazine ring containing compounds which exhibit antiviral activity and in particular human respiratory syncytial virus[HRSV] activity. The compounds are preferably anionic compounds.

This invention relates to new compounds selected from those of the general Formula I: Formula I wherein: A is C' 0C,

C,

C'

C,

C'

r r r oC' c

C,

or I C' is-

SO

3 H, OSO3H, OH, or COOH; B' is NH, NR 1 or O; R 1 is H, (C 1 -C6)lower alkyl, straight or branched, 15 wherein the carbon atoms may be optionally substituted with Cl, Br, F, OH or CN; X is Cl, F or the

Z

Nd z uw, U' is S02, -CO, or W' is:

Y-Z

Z

moiety m z Y is (CH2)n; n is 0 to 6; m is 0 to 2; Z is H, CH3, CF 3

CH

2 -(halogen), where Br, F or I, CH 2 0H, -COOH, COO(Cl-C6)lower alkyl straight or branched, -CONR 2

R

2 or halogen is Cl, Libc/02390 2

R

CN or O H and R 2 in each occurrence, is independently H or (C1-C6)lower alkyl; and the pharmaceutically acceptable salts and esters thereof.

It is understood that, in cases where m is 0, five-membered rings would be indicated for W'.

The pharmaceutical acceptable salts of the compounds of this invention are those whose cations (or anions in the case of acid addition salts) are not substantially toxic at the dosage administered to achieve the desired effect. These ions preferably do not independently possess significant pharmacological activity. Illustratively, these salts include those of alkali metals, for example, Na or K; alkaline earth metals, such as Ca or Mg; light metals of group lila including Al; and organic primary, secondary and tertiary amines or ammonia. Sodium salts are preferred.

Detailed Description of the Invention Within the group of compounds defined by Formula I, certain subgroups of compounds are broadly preferred. Broadly preferred are those compounds, or a pharmaceutically acceptable salt

C.

C.

thereof, wherein A is Especially preferred is the compound wherein C' SO 3 H, B' NH, Y CH2, Z CH 2 0H. Most preferred is the compound wherein C' SO 3 H, B' NH, Y CH2CH2, Z CONH2. Most highly and C' S3H, B NH, Y

C

preferred is the compound wherein A is and C' SOsH, B NH, Y

CH

2 CH2, Z -CONH2.

The novel compounds of the present invention may be prepared according to the following o schemes. Referring to Scheme I, condensation 2 where A and B' are hereinbefore defined with triazines having the structure 3 where X is CI, F or Br at about 0°C and pH from about 6.5 to about 7.2 gives intermediates 4. Further reaction of the compounds of the Formula 4 with compounds of formula wherein B' and W' are hereinbefore defined at a temperature of from about 45°C to about 550C and pH 6.5-7.2 followed by heating at 1000-120°C at pH6.5-7.2 gives the desired products of Formula I, above, which is referred to in the schemes below as 1.

Libc102390 Scheme 1 2

X

2 3 0OC; pH6.5-7.2

X

A

N

B -ON

H

x UW \uw 4 i 45-55*C; pH6.5-7.2 ii 100-120 C; pH 6.5-7.2 1 In an alternate route to the compounds of Formula I, as shown in Scheme II, reaction of triazine 3 where X is CI, Br or F with compounds 5, where B' and W' are hereinbefore defined at from about 5 0 C to about 5 0 C and pH 6.5-7.2 gives condensation products 6. Reaction of 6 where and W' are hereinbefore defined and X is CI, Br or F and 2 where A and B' are hereinbefore defined at from about 45 0 C to about 55 0 C and pH from about 6.5 to about 7.2 followed by heating at from about 100°C to about 120°C and pH 6.5-7.2 gives 1.

Libc/02390 Scheme II X N X H UVV NO N 3 pH6.5-7.2

X

N

_B-'U'W

X L 2 6 2 i 45-55°C; ii 100-120°C The stepwise condensation of the starting compounds 2, as shown in Scheme I, and 5 as shown in Scheme II, with triazines 3 where X is Cl, Br or F are reacted similarly in aqueous or organicaqueous media in the presence of suitable bases such as sodium or potassium hydroxides, carbonates, phosphates or bicarbonates. Phosphate buffer at pH 7.0 is preferred.

The first step of condensation is carried out in pH ranges from about 4 to about 8, preferably at a pH of from about 6.5 to about 7.2 and at temperatures from about -10°C to about 30°C, preferably 15 from about -5°C to about 5°C. The exchange of the second halogen atom of the triazine derivatives is effected in the same pH range and from about 10°C to about 70°C, preferably at a temperature of from about 45°C about 55°C. The exchange of the third halogen atom of the triazine derivatives is effected in the same pH range and at temperatures from about 80°C to about 150°C, preferably from 1000C to 1200C.

20 The exchange of the second or the third halogen atoms of the triazine derivatives of the S Formula 6 (Scheme II) is effected also in organic media in the presence of organic bases, such as trialkylamines, including triethylamine, diisopropylethylamine, or N-(lower)alkylpiperidine.

Starting materials for use in the general synthetic procedures outlined in Schemes I and II are commercially available or can be synthesized according to H. Adkins, E. F. Stembring, E. Pickering. J.

Amer. Chem. Soc., v.46, p. 1917 (1924), G.B 1 194 388 and U.S. 5 359 131. The starting compounds of Formula 2 wherein B' is oxygen can be prepared from the substituted compound of Formula 2

-NH

2 as shown in Scheme Ill.

Libcd02390

A

4

NH

2 2 HN0 2 Scheme III A{Nj N H 3 0 A-OH 2 Primary aromatic amines 7 where A defined previously are reacted with nitrous acid or with other chemical reagents (for example, organic ester of nitrous acid) for diazotization of the primary amine to yield diazonium salts 8 that are hydrolyzed to compounds of Formula 9 containing phenolic groups.

Scheme IV H 2 9 9 9* *9 9 *99 11

X=

Referring to Scheme IV, the compounds (10) of stilbene series can be transformed to the compounds (11) of bibenzyl series by catalytic reduction conditions (hydrogen Pd/C) or by related 15 reduction conditions known in the art for converting substituted stilbene compounds into bibenzyl compounds [Huang-Minlon, J. Amer. Chem. Soc. (1948) 110, 2802].

The compounds (12 and 13) of the Formula 1 with Z -COOH can be prepared by basic hydrolysis to the compounds (14) in the pH ranges from 7.5 to 10.0, preferably 8.0-8.5 and from 80 to 150°C, preferably 100 to 120°C (Scheme V): Libc/02390 7 Scheme V 12,13 0

NH

2 O0 HN 0 OH 12: R'= 0

NH

2 O0

-IN

HN 0 NH 2 4 4 *4~

I

49* 64 I.*4

I

4 St 0

R=

m-Aminobenzamides 17 where U' is -SO2 or -00, Y and Z are hereinbefore defined, are synthesized according to the Scheme V1.

Libc/O2390 8 Scheme VI U Y'Z O=N -N 8

\I

14 base c Y UU-- N O=N

Y

I H 2 /Pd H 2

N

16 17 Imides 15 are transformed into m-nitrosulfonates 16, using a suitable variant of the amidation or tosylation methods W. Greene, P. G. M. Wuts, Protective Groups in Organic Chemistry, John Wiley &Sons, New York 1991, pp.349-37 9 The nitro compounds 16 are then reduced to the aminocompounds 17 by catalytic hydrogenation (hydrogen -Pd/C).

Piperazine derivatives 21-22, where Z is -COOH or -CONH2, are synthesized according to the Scheme VII from 3,5-piperidinedicarboxylic acid 18. Reduction of the aromatic ring and mnitrotosylation of the piperidine derivative 19 leads to the compound 20 which is either reduced to a carboxyl containing compound 21 or converted into a carbamoyl containing compound 22 by 15 esterification and ammonolysis.

o* Libc/02390 Scheme VII 0

O=N

H

2 /Pt

Z

O^S-N

11 H0 H 2 /Pd H2N Z 21,22 H2/Pd 0 oOH

II

0

P-N-

O=N

OH

II O 0 0 I CH 3

N

3 n c 0

OH

Q- -N O=N

OH

II

0

NH

3 /EtOH

II

u 24 23 This invention also includes the methods of treating a mammal, preferably a human, experiencing a viral infection, the methods comprising administering to the mammal so afflicted one or more of the compounds of this invention, and/or one or more pharmaceutically acceptable salts of the compounds. Viral infections treatable using the compounds and methods of this invention include those infections brought on by human respiratory syncytial virus, herpes simplex viruses, human cytomegalovirus, and influenza viruses, particularly parainfluenza virus 3.

The compounds of this invention may be administered to the mammal in need thereof in any manner prescribed, including intranasally, orally, topically, transdermally, parenterally and intraperitoneally. It is understood that treating the mammal, preferably the human, will require varying dosages of active compound and varying regimens of treatment depending upon various factors including the age, sex, size, general health and extent of disorder seen in the given individual, the dosage and regimen to be determined by the proper medical practitioner. An antivirally effective Libc/02390 amount of the compounds herein to be administered will generally range from about 0.5mg/kg to about 500mg/kg of animal body weight in one or more doses per day. The doses are preferably given in divided doses two to four times a day, or in a sustained release form. For most large mammals the total daily dosage is from about 1 to 100mg, preferably from about 2 to 80mg. Dosage forms suitable for internal use comprise from about 0.5 to 500mg of the active compound in intimate admixture with a solid or liquid pharmaceutically acceptable carrier. This dosage regimen may be adjusted to provide the optimal therapeutic response. For example, several divided doses may be administered daily or the dose may be proportionally reduced as indicated by the exigencies of the therapeutic situation.

The compounds of this invention may be given with or without other antiviral agents.

The present invention also includes pharmaceutical compositions which may be used with these methods of treatment. The pharmaceutical compositions may comprise one or more of the compounds of this invention, or pharmaceutically acceptable salts thereof, and one or more pharmaceutically acceptable carriers, for instance, diluents, excipients, fillers, binders, flavoring agents, etc. For oral administration, the compositions containing the compounds of this invention include solid or liquid compositions, such as capsules, troches, lozenges, pills, tablets, powders, melts, solutions, suspensions and emulsions. The solid forms may be encapsulated by hard or soft gelatin capsules and may contain commonly used pharmaceutical acceptable excipients, fillers, adjuvants, diluents, lubricants, disintegrants, suspending or stabilizing agents, and binding agents including, but not limited to, magnesium stearate, sodium lauryl sulfate, microcrystalline cellulose, polyvinylpyrrolidone, gelatin, alginic acid, acacia gum, sodium citrate, complex silicates, calcium carbonate, glycine, dextrin, sucrose, sorbitol, calcium phosphate, dicalcium phosphate, calcium sulfate, lactose, kaolin, mannitol, sodium chloride, talc, dry starch corn, potato or tapioca starch) and powdered sugar. The formulations may also include antioxidants, for example, vitamin E, ascorbic acid, BHT and BHA. Such pharmaceutical preparations may contain, for example, from about ~25 0.05 up to about 90% of the active ingredient in combination with the carrier, more usually between about 5% and 60% by weight.

Suitable excipients for liquid oral formulations include diluents such as water, and alcohols such as ethanol, benzyl alcohol, and polyethylene alcohols, with or without a surfactant, suspending agent o" or emulsifying agent. Dispersions can be prepared in glycerol, liquid polyethylene glycols and mixtures S 30 thereof in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.

The compounds of this invention may also be administered parenterally as an injectable dosage form in a physiologically acceptable diluent such as sterile liquids or mixtures thereof water, including water, saline, aqueous dextrose and other pharmaceutically acceptable sugar solutions, alcohols such as ethanol, isopropanol, or hexadecyl alcohol, glycols such as propylene glycol or polyethylene glycol, glycerol ketals such as 2,2-dimethyl-1,3-dioxolane-4-methanol, ethers such as poly(ethyleneglycol) 400, a pharmaceutically acceptable oil, fatty acid, fatty acid ester or glyceride, or an acetylated atty acid glyceride with or without the addition of a pharmaceutically acceptable surfactant, such as a soap or detergent, suspending agent such as pectin, carbomers, methylcellulose, hydroxypropylmethylcellulose, or carboxymethylcellulose, an emulsifying agent or Libc/02390 pharmaceutical adjuvants. In all cases, the form must be sterile and must be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of micro-organisms such as bacteria and fungi.

Pharmaceutically acceptable oils which are useful in the formulation herein include those of petroleum, animal, vegetable or synthetic origin, including peanut oil, soybean oil, sesame oil, cottonseed oil, olive oil, sunflower oil, petrolatum, and mineral oil. Fatty acids which may be used include oleic acid, stearic acid, and isostearic acid, while the fatty acid esters useful herein may include ethyl oleate and isopropyl myristate. Suitable soaps include fatty acid alkali metal, ammonium, and triethanolamine salts. Acceptable detergents include cationic detergents, for example, dimethyl dialkyl ammonium halides, alkyl pyridinium halides, and alkylamine acetates and anionic detergents, such as alkyl, aryl, and olefin sulfonates, alkyl, olefin, ether and monoglyceride sulfates, and sulfosuccinates. Useful non-ionic detergents may include fatty amine oxides, fatty acid alkanolamides and polyoxyethylenepolypropylene copolymers. Amphoteric detergents may include alkyl-betaaminopropionates and 2-alkylimidazoline quaternary salts, and mixtures thereof.

The parenteral compositions of this invention preferably will contain from about 0.5 to about by weight of the active compounds described herein in solution. The parenteral formulations in the form of sterile injectable solutions or suspensions will also preferably contain from about 0.05% to about 5% suspending agent in an isotonic medium. Buffers and preservatives may be added. A suitable surfactant may also be added. These surfactants may include polyethylene sorbitan fatty acid esters, such as sorbitan monooleate, and the high molecular weight adducts of ethylene oxide with a hydrophobic base, formed by the condensation of propylene oxide with propylene glycol.

The pharmaceutical compositions of this invention also include those useful for topical and transdermal administration. These formulations may include the dermal application of the compounds of this invention in a solvent system noted for enhancing transdermal absorption, including ethanol or dimethylsulfoxide, with or without other excipients. Preferably the topical and transdermal compositions herein comprise the addition of the compounds of this invention into a patch of the reservoir and porous membrane type or in a patch of the solid matrix variety. Transdermal patches of these types are described in US. 3 742 951, 3 797 494, 3 996 934, 4 031 894, 4 573 996, and 4 956 171.

The intranasal formulations and administrations of this invention may be administered as intranasal drops or as an intranasal spray, such as an inhalant. The formulations may include any combination of pharmaceutically acceptable components which are useful in intranasal administration, including sterile water, saline, a stabilizing agent, such as polyethylene glycol having a molecular weight in the range of from about 200 to about 7500, or mixtures thereof.

35 The compounds of this invention can be used to treat viral infections either prophylactically or therapeutically. For prophylactic use, a compound could be administered intranasally as a spray or a mist or as drops on a daily basis to prevent infection by virus, in particular by RSV. The length of time during which treatment would be carried out will reflect the likelihood of infection by a specific viral agent and may be affected by epidemic situations and the risk factors displayed by a given patient.

Compounds delivered in these ways may range in concentration from 0.1 to 30mg/mL, the volume Libc/02390 delivered ranging from 0.1 to lmL/nostril. Alternatively, the compounds can be delivered systemically by oral or intravenous dosing. For therapeutic use, the compounds may be delivered topically to the lungs as an aerosol or could be delivered systemically by oral or intravenous dosing. Such dosing may be administered for a period of time necessary to control the viral infection.

The compounds of this invention and their preparation can be understood further by the following non-limiting examples. The reaction intermediates, products and potential by-products are analyzed by thin-layer chromatography (TLC) on silica gel using of propan-2-ol-ethyl acetate-4% ammonia in water v/v) as solvent, and reversed-phase high-performance liquid chromatography (RP-HPLC) [column: Vydac C18 (4.6mm x 25cm), 5mm; mobil phase: A-acetonitrile, B-0.1M ammonium acetate in water (pH5.5), C-methanol; gradient: from mixture A-B-C (20:65:15, v/v) to mixture A-B-C (20:55:25, v/v) in 10 minutes]. Preparative RP-HPLC is performed on a Rainin gradient HPLC system using a Vydac C18 Peptide/Protein preparative column (10mm, 22mm x mobile phase: A-methanol-0.1M ammonium acetate in water (pH 5.5) v/v) B-methanolacetonitrile gradient: 18-32% B in 10 minutes.

Example 1 4.4'-bis[4,6-di[3-aminophenyl-N,N-bis(2-carbamo-ethyl)sulfonylimino]-1.3.5-triazin-2-ylamino]-stilbene- 2,2'-disulfonic acid, disodium salt A solution of cyanuric chloride (3.87g, 21mmole) in dioxane (25mL) was added to phosphate buffer (100mL, 0.3 M, pH 7) with stirring at A solution of 4,4'-diaminostilbene-2,2'-disulfonic acid (3.70g, 10mmole) in iN NaOH (20mL) was then added over a 20 minute period and the pH maintained at 6.5-7.2 by addition of 1N NaOH. Stirring was continued for a further 1h at the same temperature and pH while the reaction was monitored for completeness by analytical HPLC (yield A solution of 3-aminophenyl-N,N-bis(2-carbamoylethyl)sulfonylimine (13.20g, 42mmole)(GB.

1,194,388) in dimethylsulfoxide(DMSO) (100mL) was added over a 20 minute period and the 25 temperature raised to 50 0 C. Stirring was continued for 2.5h at this temperature and then for 40h at 100-110°C with the pH maintained at 6.6-7.2 by addition of 1N NaOH. Once the reaction was complete as determined by analytical HPLC (or TLC), the mixture was cooled to 20 0 C and acidified with 5.6N hydrochloric acid to pH2. Sodium chloride (60mL, 4M) was added and the precipitated i' product filtered, redissolved in a minimum volume of water by addition of 1N NaOH to pH7 and re- 30 precipitated by the addition of sodium chloride. The precipitate was filtered, washed with cold water propan-2-ol (40mL), acetone (60mL) and dried under vacuum at 50 0 C. Yield 13.0g m.p. >250 0 C dec; UV(water): 273nm (logs 5.11), 350nm (logs 4.89); MS(ES) M- 2 887.9; MW 1821.2.

Example 2 35 4.4'bis[4,6-di[3-aminophenyl-N,N-bis(2-carbamoyl-ethyl)sulfonylimino]-1.3.5-triazin-2.ylaminol-stilbene- 2,2'-disulfonic acid, disodium salt A suspension of 4,4'-diaminostilbene-2,2-disulfonic acid, disodium salt (1.21g, 2.92mmole) and 2-chloro-4,6-di[3-aminophenyl-N,N-bis(2-carbamoylethyl)-sulfonylimino]-1,3,5-triazine (Example 6) (4.47g, 6.44mmole) in sulfolane (70mL) and diisopropylethylamine (1.5mL) was warmed in a sealed thick-walled glass tube to 115 0 C. Warming was continued for 40h. Once the reaction was complete Libc/02390 as determined by analytical HPLC (or TLC), the mixture was cooled to 30'C, 1IN NaOH (6.6mL) was added and the product precipitated by the addition of propan-2-ol (200mL). The precipitate was filtered, washed with acetone (3OmL) and redissolved in a minimum volume of water and reprecipitated by the addition of ethanol. The precipitate after filtration was washed with ether (3OmQ) and dried under vacuum at 50'C. Yield 4.84g Example 3 4.41bis[4,6-di[3-aminophenylN,N-bis(2.carbamoyl-ethyl)sulfonyliminol1i .3.5-triazin-2.ylamirio]-stilbene- 2,2'-disulfonic acid, disodium salt The title compound was prepared using the general conditions of Example 1, where 2,4,6trifluoro-l ,3 ,5-triazine (304mg, 2.25mmole) was reacted with 4,4'-diaminostilbene-2,2'-disulfonic acid (455mg, 1.1 Ommole) for 1 h, and then with 3-aminophenyl-N ,N-bis(2-carbamoylethyl)sulfonylimine (1.45g, 4.62mmole)(GB. 1,194,388), followed by purification, to give the desired compound (1.21g,61%). The compounds synthesized by procedures of Examples 1-3 are identical according to HPLC, UV and mass-spectra.

Example 4 4.4'bis[4,6-di[3aminophenylN,Nbis(2-carbamoylethysl)foflimino]- .3.5-triazin.2-ylamino]-biphelyl- 2,2'-disulfonic acid, disodium salt The title compound was prepared according to the procedure of Example 2 using 4,4'diaminobiphenyl-2,2'-disulfonic acid, disodium salt (1 .02g, 2.62mmole) and 2-chloro-4,6-di[3aminophenyl-N ,N-bis(2-carbamoylethyl)sulfonilinmino]-l ,3,5-triazine (Example 6) (4.26g, 5.76mmole).

in 6OmL DMSO-sulfolane v/v) and diisopropylethylamine (1 .2mL). The product was obtained as a colorless solid (3.90g, m.p. >25000 dec; UV(water): 272nm (logs 4.86); MS(ES) (mlz): M- 2 874.1; MW 1794.2.

25Example 5 4,4'-bis[4,6-di[3-aminophelN,Nbis(2hydroxyethyI)SUlfonylimiflo]H.3.5.triazin-2-ylamiflo]-stilbefle-2, 2 disulfonic acid, disodium salt The title compound was prepared according to the procedure used to prepare Example 2 using 4,4'-diaminostilbene-2,2'-disulfonic acid (28mg, 0.O75mmole) and 95mg (0.l5mmole) of 2-chloro-4,6di[3-aminophenyl-N ,N-bis(3-hydroxyethy)sulfonilimlinoI-l ,3,5-triazine (Example 6) in sulfolane and diisopropylethylamine (35mL) at 11000 for 18h. The cooled reaction mixture was mixed with deionised water (4OmL) and the solution of crude product was purified by preparative RP-HPLC.

Fractions containing the desired product are combined, organic solvents are evaporated under vacuum (10mm Hg) at 3000, and the product was concentrated, desalted and isolated via octadecyl (018) cartridge (900mg, obtained from Burdick Jackson). The cartridge was washed with water (3OmQ) and the product reextracted with methanol (5OmL). Evaporation of the methanol gave 12mg of the desired product; m.p. >25000 dec; UV(water): UV(water): 273nm (logs 4.97), 350nm (logs; 4.81); MS(ES) M- 2 779.0; MW 1604.0.

Example 6 2-chloro-4,6-di[3-amiflophelyl-N,N,-bis(2-carbamoyi-ethyl)sulfonyliminol,3,5.triazine A solution of cyanuric ch loride (4.2g, 22.7mmole) in dioxane (55mL) was added dropwise with stirring to phosphate buffer (12OmL, 0.3M, pH7) and crushed ice (l0g) at -20C0OOC. To the resulting Libc102390 fine suspension was added dropwise over a 30 minute period a solution of 3-aminophenyl-N,N-bis(2carbamoylethyl)sulfonylimine (15.0g, 47.8mmole) (GB. 1,194,388) in N,N-dimethylformamide (125mL) and the pH maintained at 6.5-7.2 by addition of 1N NaOH. Stirring was continued for a further 1h pH7.0) and the temperature raised to 55°C. Stirring was continued for 2.5h while the reaction is monitored for completeness by analytical HPLC (or TLC). Once the reaction was complete as determined by analytical HPLC (or TLC), the mixture was cooled to 0°C, water (160mL) was added and the product precipitate filtered, washed with cold water (50mL), acetone (50mL) and dried under vacuum at 40°C. Yield 14.5g m.p. >250°C dec; UV(DMSO): 282nm (logs 4.77); MS(CI) MH 740.1.

Example 7 4-[4,6-di[3-aminophenyl-N,N-bis(2-carbamoylethyl)sulfonylimino]-1,3,5-triazin-2-ylamino]-4'-[4-chloro- 6 aminophenyl-N,N-bis(2-carbamoylethyl)sulfonylimino]-1,3,5-triazin-.2ylamino]-stilbene-2,2'-disulphonic acid, disodium salt A solution of cyanuric chloride (205mg, 1.12mmole) in dioxane (1.5mL) was added to phosphate buffer (10mL, 0.3M, pH7) with stirring at A solution of 4,4'-diaminostilbene-2,2'disulfonic acid, disodium salt (225mg, 0.54mmole) in phosphate buffer (3mL) was added and the pH maintained at 6.5-7.2 by addition of 1N NaOH. Stirring was continued for a further 1h at the same temperature and pH while the reaction was monitored for completeness by analytical HPLC (yield A solution of 3-aminophenyl-N,N~bis(2-carbamoylethyl)sulfonylimine (700mg, 2.23mmole) in N,N-dimethylformamide (10mL) was added over a 10 minute period and the temperature raised to Stirring was continued for 3h at this temperature and 20h at 100-110°C with the pH maintained at 6.6-7.2 by addition of 1N NaOH. Once the reaction was complete as determined by analytical HPLC, the mixture was cooled to 20°C and acidified with 5.6N hydrochloric acid to pH2, sodium chloride (5mL, 4M) was added and the product precipitate filtered, washed with cold water (2mL), and 25 dried under vacuum at 40°C. Two portions of a solution of crude product (about 1g) in deionised water (250mL) were purified separately by preparative RP-HPLC. Fractions containing the desired product were combined, organic solvents were evaporated under vacuum (10mm Hg) at 30°C, and the product was concentrated, desalted and isolated via octadecyl (C18) cartridge (900mg, obtained from Burdick Jackson). The cartridge was washed with water (30mL) and the product reextracted with 30 methanol (50mL). Evaporation of the methanol gives 268mg of the desired product, m.p.

>2500C dec; UV(water): 273nm (logs 4.83), 350nm (logs 4.44); MS(ES) M- 2 749.15; MW 1544.3.

Example 8 4.4'-bis[4-chloro-6-[3-aminophenyl-N,N-bis(2-carbamoylethyl)sulfonylimino]-1,3,5.triazin-2-ylamino]stilbene-2,2'- acid. disodium salt A solution of cyanuric chloride (0.91g, 4.95mmole) in dioxane (8mL) was added to phosphate buffer (30mL, 0.3M, pH7) with stirring at A solution of 4,4-diaminostilbene-2,2'-disulfonic acid, disodium salt (1.00g, 2.41mmole) in deionised water (9mL) is added and the pH maintained at 6.5-7.2 by addition of 1N NaOH. Stirring was continued for a further 1h at the same temperature and pH while the reaction was monitored for completeness by analytical HPLC (yield A solution of 3aminophenyl-N,N-bis(2-carbamoylethyl)sulfonylimine (1.57g, 5.00mmole) in N,N-dimethylformamide Libc/02390 was added over a 10min period and the temperature raised to 50°C. Stirring was continued for 6h at this temperature with the pH maintained at 6.6-7.2 by addition of 1N NaOH. Once the reaction was determined complete by analytical HPLC (or TLC), the mixture was cooled to 00C and acidified with 5.6N hydrochloric acid to pH2. Sodium chloride (5mL, 4M) was added and the product precipitate filtered, washed with cold water (8mL), acetone (20mL) and ether (20mL), and dried under vacuum at 40°C. Yield 2.37g m.p. >300 0 C dec; UV(water): 273nm (logs 4.57). 350 (logs 4.42); MS(ES) M- 2 610.1; MW 1266.2.

Example 9 and Example 4-[4,6-di[3-aminophenyl-N,N-bis(2-carbamoylethyl)sulfonylimino]-1,.3,5-triazin-2-ylamino]-4'-[4-[3aminophenyl-N,N-bis(2-carbamoylethyl)sulfonylimino]-6-[3-aminophenyl-N-( 2 -carbamoylethyl)-N'-( 3 propionic acid)]-1,3,5.triazin-2-ylamino]-stilbene-2,2'-disulphonic acid, disodium salt and 4-[4,6-di[3aminophenyl-N,N-bis(2-carbamoylethyl)sulfonylimino]-1,3,5-triazin-2-ylamino]--4'-[4-[3-aminophenyl- N,nN-bis(2-carbamoylethyl)sulfonylimino]-6-[3-aminophenyl-N,N-bis( 3 -propionic acid)]-1.3.5-triazin-2ylamino]-stilbene-2,2'-disulfonic acid, disodium salt 1s To a solution of Example 1(200mg, 0.11mmole) in deionised water (20mL) was added 1N NaOH (300mL) and the mixture heated to boiling. The reaction mixture was kept warm for 3h, cooled to O°C and acidified with 5.6N hydrochloric acid to pH2. Sodium chloride (10mL, 4M) was added and the product precipitate filtered, washed with cold water (2mL), and dried under vacuum at 400C. A solution of crude product (about 180mg) in distilled water (125mL) was purified by preparative RP- HPLC. Fractions containing the desired product (Example 9) are combined, organic solvents are evaporated under vacuum (10mm Hg) at 30°C, and the product was concentrated, desalted and isolated via octadecyl (C18) cartridge (900mg, obtained from Burdick Jackson). The cartridge was washed with water (30mL) and the product reextracted with methanol (50mL). Evaporation of the methanol yielded 24mg of the desired product., m.p. >250°C dcc; UV(water): 273nm (logs 5.02), 350nm (logs 4.76); MS(ES) (mlz): M- 2 888.3; MW 1822.6.

The second product (Example 10) is isolated from the other fractions containing the desired compound (Example 10) by RP-HPLS-separation while using the same procedure of concentration, desalting and isolation via octadecyl (C18) cartridge described in Example 9. Yield 18mg m.p.

>250°C dec, UV(water) 273nm (logs 4.92), 350 (logs 4.69); MS(ES) M- 2 889.2; MW 1824.4.

30 Example 11 4.4'-bis[4,6-di[3-aminophenyl-N,N-bis(2-carbamoylethyl)sulfonylimino]-1,3,5-triazin-2-ylamino]-bibenzyl- 2.2'-disulfonic acid, disodium salt A solution of Example 1(30mg) in 30mL water-methanol v/v) was stirred with palladium on charcoal (10% Pd) in a hydrogen atmosphere for 5 days. The reaction mixture was filtered and the solvent evaporated. The product was isolated by preparative HP-HPLC. The fractions containing the desired product were combined, organic solvents evaporated under vacuum (10mm Hg) at 30°C, and the product isolated via octadecyl (C18) cartridge (300mg, obtained from Burdick Jackson). The cartridge was washed with water (30mL) and the product reextracted with methanol Evaporation of the methanol left 16mg of the desired product; m.p. >250°C dec; UV(water):274nm (logs 4.96); MS(ES) M- 2 889.2; MW 1824.4.

Libc/02390 Example 12 4.4'-bis[4,6-di[4-aminophenyl-N,N-bis(2-carbamoylethyl)suIfonylimino]-1,3,5-triazin-2-ylamino]-stilbene- 2,2'-disulfonic acid, disodium salt The title compound was prepared according to the procedure used for the synthesis of Example 2 using 4,4'-diaminostilbene-2,2-disulfonic acid (48mg, 0.125mmole) and 2-chloro-4,6-di[4aminophenyl-N,N-bis(2-carbamoylethyl)sulfonylimino]-1 ,3,5-triazine (Example 18) (230mg, 0.310mmole) in dimethylsulfoxide (12mL) and diisopropylethylamine (260mL). The product was obtained as a colorless solid (173mg, m.p. >2500C dec; UV(water): 296nm (logs 4.94), 352nm (logs 4.57); MS(ES) (mlz): M- 2 883.2; MW 1822.4.

Example 13 4.4'-Bis~4.6-di[4-aminophenyl-N,N-bis(2-carbamoylethy)sulfonylimino]-l ,3,5-triazin-2-ylamino]-biphenyl- 2.2'-disulfonic acid, disodium salt The title compound was prepared according to the procedure used for the synthesis of Example 2 using 4,4'-diaminobiphenyl-2,2'-disulfonic acid, disodium salt (113mg, 0.33mmole) and 2-chloro-4,6di[4-aminophenyl-N,N-bis(2-carbamoylethyl)sulfonylimino]-l ,3,5-triazine (Example 18) (535mg, 0.72mmole) in 25mL DMSO-sulfolane v/v) and diisopropylethylamine (350mL). The product was obtained as a colorless solid (450mg, m.p. >2500C dec; UV(water): 296nm (logs 5.13 MS(ES) (mlz): M- 2 874.4; MW 1794.8.

Example 14 2-chloro-4,6-di(3'-sulfoamidoanilino)-1 A solution of cyanuric chloride (2.37g, 12.9mmole) in acetone (25mL) was added dropwise with stirring to phosphate buffer (60mL, 0.3M, pH7) at -2-00C. To the resulting fine suspension was added dropwise over a 20 minute period, a solution of m-sulfanilamide (4.35g, 25.3mmole) in acetone and the pH maintained at 6.5-7.2 by addition of 1N NaOH, Stirring was continued for a further 30min (0OC, pH7.0) and the temperature raised to 550C. Stirring was continued for 3h, while the reaction was monitored for completeness by analytical HPLC (or TLC). Once the reaction was complete as determined by analytical HPLC (or TLC), the mixture was cooled to 00C, water (100mL) was added and the product precipitate filtered, washed with cold water (50mL), dried on the filter, and purified by dissolving in hot acetone (40mL) and precipitation with benzene (150mL). The product was 30 filtered, washed with propan-2-ol (20mL) and ether (30mL), dried under vacuum at 300C. Yield 3,72g m.p. >250'C dec; UV(MeOH): 277nm (logs 4.68); MS(Cl) (mlz): MH 456.0.

Example 4.4'-bis[4,6-di[3aminophenylsulfonylamido]-1,3,5-triazin-2-ylamino]-stilbene-2,2'-disulfonic acid, disodium salt The title compound was prepared according to the procedure used for the synthesis of Example "2 using 4,4'-diaminostilbene-2,2'-disulfonic acid, disodium salt (216mg, 0.52mmole) and 2-chloro-4,6di(3'-sulfoamidoanilino)-1,3,5-triazine (Example 14) (524mg, 1.15mmole) in sulfolane (15mL) and diisopropylethylamine (250mL,) for 40h at 115'C. After cooling to 200C, the reaction mixture was diluted with ether (100mL) and 1N NaOH (2.08mL) added. After centrifugation, the residue was stirred With propan-2-ol (50mL) until crystallization was complete. The solid was centrifuged, washed with Libcl02390 methanol (2x30mL) and ether (2x30mL), and dried under vacuum at 30'C. Yield (570mg, 92 m.p.

>2500C dec; UV(water): 273nm (logs 4.76), 350nm (logs 4.52).

Example 16 4,4'-bis[4,6-di[2-aminophenylsulfonylimido]-1 ,3,5-triazin-2-ylamino]-biphenyl.2,2'disulfonic acid, disodium 6 salt The title compound was prepared according to the procedure used for the synthesis of Example 2 using 4,4'-diaminobiphenyl-2,2'-disulfonic acid (208mg, 0.60mmole).and 2-chloro-4,6-di(3'sulfoamidoanilino)-1,3,5-triazine (Example 14) (607mg, 1.33mmole) in sulfolane (20mL) and diisopropylethylamine (850mL,) for 40h at 115 0 C. After cooling to 200C, the reaction mixture was diluted with ether (150mL) and 1N NaOH (2.40mL) added. After centrifugation, the residue was stirred with propan-2-ol (50mL) until crystallization was complete. The solid was centrifuged, washed with methanol (40mL) and ether (2x20mL), and dried under vacuum at 300C Yield (505mg, m.p.

>2500C dec; UV(water): 276nm (logs 4.93).

Example 17 4,4'-dihydroxybiphenyl-2,2'-disulfonic acid To a suspension of 4,4'-diaminobiphenyl-2,2'-disulfonic acid (4.44g, 12.9mmole) in water was added a solution of sodium carbonate (8.6mL, 1.5 M) by warming until a clear solution was obtained. After cooling to 50C, to this solution was added a solution of sodium nitrite (1.78g, 12.9mmole) in water (3mL). The resulting solution was added dropwise with stirring to a solution of concentrated sulfuric acid (3.8mL) in water (13mL) and crushed ice (10g) at -5 -2oC. Stirring was continued for a further 30 minutes, and the reaction mixture was warmed to 650C. The temperature was maintained until nitrogen evolution takes place. The mixture was cooled to 1000 and neutralized with dry sodium carbonate, and the solution evaporated to a residue. The residue was dried under vacuum at 500C, mixed with ethanol (225mL) and filtered. The filtrate was evaporated and dried under S 25 vacuum at 400C. Yield 3.88 m.p. >2500C dec; UV(0.05N NaOH): 252nm (logs 4.16), 311nm (logs 3.40); MS(ES) 345.0; MW 346.0.

Example 18 2-chloro-4,6-di[4-aminophenyl-N,N-bis(2-carcamoyiethyl)sulfonylimino -1,3,5-triazine The title compound was prepared according to the procedure used for the synthesis of Example 6 using 4-aminophenyl-N,N-bis(carbamoylethyl)sulfonylimine (3.3g, 10.5mmole) (GB. 1 194 388) and cyanuric chloride (1.0g, 5.4mmole). The product was obtained as a colorless solid (3.5g, m.p.

>2500C dec; UV(DMSO): 303nm (logs 4.84); MS(CI) (mlz): MH 740.1.

Examplel9 4 4 -bis[4,6-di[3-aminophenyl-N,N-bis(2-carbamoylethyl)sulfonylimino-1 ,3,5-triazin-2-yloxy]-biphenyl- 2 2 disulfonic acid, disodium salt A suspension of 4,4'-dihydroxybiphenyl-2,2'-disulfonic acid (Example 17) (131mg, 0.34mmole) and 2-chloro-4,6-di[3-aminophenyl-N,N-bis(2-carbamoylethyl) sulfonylimino]-1,3,5-triazine (Example 6) (550mg, 0.74mmole) in sulfolane N,N-dimethylformamide (15mL, 2:1 v/v) and NaOH (IN, 1.48mL) was warmed in a sealed thick-walled glass tube to 1150C. Warming was continued for 50h. Once the reaction was complete as determined by analytical HPLC (or TLC), the mixture was cooled to Libc102390 and the product precipitated by addition of propan-2-ol (70mL), washed on filter with cold ethanol and dried under vacuum at 400C. Two portions of a solution of crude product (about 0.6g) in deionised water (200mL) were purified separately by preparative RP-HPLC. Fractions containing the desired product were combined, organic solvents evaporated under vacuum (10mm Hg) at 300C, and the product concentrated, desalted and isolated via octadecyl (C18) cartridge (900mg, obtained from Burdick Jackson). The cartridge was washed with water (30mL) and the product reextracted with methanol (50mL). Evaporation of the methanol gives 171mg of the desired product; m.p.

>2500C dec; UV(water): 272nm (logs 4.86); MS(ES) M- 2 875.2; MW 1796.4.

Example 5 .5'-dimethyl-4,4'-bis[4,6-di[3-aminophenyl-N,N-bis(2-carbamoylethyl)sulfonylimino]-1,3,5-triazin-2ylamino]-biphenyl-2,2'-disulfonic acid, disodium salt The compound was prepared according to the procedure used for the synthesis of Example 2 using 5,5'-dimethyl-4,4'-diaminobiphenyl-2,2'-disulfonic acid, disodium salt (206mg, 0.49mmole) and 2-chloro-4,6-di[3-aminophenyl-N,N-bis(2-carbamoylethyl)-sulfonylimino]-1 ,3,5-triazine (Example 6) (805mg, 1.08mmole) in 30mL DMSO-sulfolane v/v) and diisopropylethylamine (200mL) for 50h at 115*C. The product after HPLC-purification was obtained as a colorless solid (191mg, 22%); m.p.>250'C dec; UV(water): 274nm (logs 5.08); MS(ES)

M-

2 8883.3; MW 1822.6.

Example 21 2-chloro-4,6-di[3-aminophenyl-N,N-bis(3-hydroxyethyl)sulfonylimino]1 A solution of cyanuric chloride (135mg, 0.73mmole) in dioxane (55mL) was added dropwise with stirring to phosphate buffer (120mL, 0.3M, pH7) and crushed ice (10g) at from to OC. To the resulting fine suspension was added dropwise over a 20 minute period a solution of 3-aminophenyl- N,N-bis(2-hydroxyethyl) sulfonylimine (365mg, 1.40mmole) in N,N-dimethylformamide (125mL) and the pH maintained at 6.5-7.2 by addition of 1N NaOH. Stirring was continued for a further 1h (00C, 25 pH7.0) and the temperature raised to 550C. Stirring was continued for 2.5h, while the reaction was :i monitored for completeness by analytical HPLC (or TLC). Once the reaction was complete as determined by analytical HPLC (or TLC), the mixture was cooled to 00C, water (160mL) was added and the product precipitate filtered, washed with cold water (50mL), acetone (50mL) and dried under vacuum at 400C. Yield 395mg m.p. >250*C dec; UV (DMSO): 284nm.

Example 22 4,4'-Bis-4,6-di[3-aminophenyl-N,N-bis(2-hydroxyethyl)sulfonylimino]-1 .3.5-triazin-2-ylamino]-biphenyl-2, 2 disulfonic acid, disodium salt This compound was prepared according to the procedure used for the synthesis of Example 2 using 4,4'-diaminobiphenyl-2,2'-disulfonic acid (28mg, 0.08mmole) and 115mg (0.17mmole) of 2chloro-4,6-di[3-aminophenyl-N,N-bis(3-hydroxyethyl)sulfonylimino]-1 ,3,5-triazine (Example 21) in sulfolane (10mL) and diisopropylethylamine (25mL) at 1100C for 18h. The product after HPLC purification was obtained as a colorless solid (16mg, m.p. >2500C dec; UV(water): 272nm (logs 4.88); MS(ES)

M-

2 776.0; MW 1534.0.

Libc/0O2390 19 Example 23 9 9 .dioxo.

2 ,7-bis[4,6di[3amilophelyl.N,N-bis(2-carbamoylethyl)sulfonyliminoi]i ,3,5-triazin-2-ylaminoldjbenzothiophen3,6-disulfoflic acid, disodium salt The title compound was prepared according to the procedure used for the synthesis of Example 1 using a solution of cyanuric chloride (59mg, 0.32mmole), 9,9-dioxo-2,7-diamino-dibenzothiophen- 3,6-disulfonic acid (77mg, 0.l6mmole) and 2-chloro-4,6-di-[4-aminophenyl-N,N-bis(2carbamoylethyl)sulfonylimino)-1, 3 ,S-triazine (Example 6) (300mg, 0.96mmole) in dioxane (l5ml-) and phosphate buffer (2Oml-, 0.3M, pH 7) at 11000 for 18h. The product after HPL0C-purification was obtained as a colorless solid (64mg, 11 m.p. >25000 dec; MS(ES)

M-

2 905.1; MW 1858.2.

Example 24 I ,3,5-triazin-2ylamilstilbefle.2,2'disulfoflic acid, disodium salt The compound was prepared according to the procedure used for the synthesis of Example 2 using 4,4'-diaminostilbene-2,2'-disulfonic acid (20mg, 0.OS4mmole) and 112mg (0.l2mmole) of 2,4dichloro-6-[4-amino-N'-[3-aminophenyl-N ,N-bis(2-carbamoylethyl)sulfonyliminoI-benzamideI-I13,5triazine (Example 26) in sulfolane (2OmL-) and diisopropylethylamine (38mL) at 11000C for 18h. The product after HPLC purification was obtained as a colorless solid (20mg, m.p. >250'0 dec; UV(water): 295nm (logs 4.54), 350nm (logs 4.39); MS(ES)

M-

2 727.1; MW 1482.2.

Example I ,3,5-triazin-2-ylamilo]-stilbelG.2,2'-disulfoflic acid, disodium salt The title compound was prepared according to the procedure used for the synthesis of Example 2 using 4,4'-diaminostilbene-2,2'-disulfonic acid (20mg, 0.O54mmole) and 203mg (0.22mmole) of 2- *chloro4,6-di [4-amino-N'-[3-aminophenyl-N N-bis(2-hyd roxyethyl)su lfonylimino-benzensu Ifon amid e]- 1,3,5-triazine (Example 27) in sulfolane (2OmL-) and diisopropylethylamine (38mL) at 11000C for 18h.

The product after HPL-C-purification was obtained as a colorless solid 30mg, m~p. >2500C dec; UV(water): 295nm (logs 5.21), 350nm (logs 4.69); MS(ES)

M-

2 1089.3; MW 2226.6.

Example 26 2 4 -dichloro.l4-amino-NP[3amilophelN,N-bis(2carbamoylethylsulfonylimino]-benzamide]l 30 triazine A solution of cyanuric chloride (56mg, 0.3mmole) in dioxane (lmL) was added with stirring to phosphate buffer (l2mL, 0.3 M, pH 7) at -20C. To the resulting suspension was added dropwise a solution of 4-amino-N'-[3-aminophenyl-N ,N-bis(2-carbamoylethyl)sulfonyliminolbenzamide (130mg, 0.3mmole) in N,N-dimethylformamide (2mL-) and the pH maintained at 6.5-7.2 by addition of 1N NaOH. Stirring was continued for a further 1h (000, pH then water (2Oml-) was added and the product precipitate filtered, washed with cold water (l0mL-), acetone (5mL) and dried under vacuum at 2000 to give 108mg of the desired product; m.p. >25000 dec; UV (DMS0) 285nm.

Example 27 2 .chloro.

4 6 .di[ 4 -amilo-N[3amilophelN,N-bis(2hydroxyethyl)sulfonylimino-benzensulfonamide]- 1,3,5-triazine The title compound was prepared according to the procedure used for the synthesis of Example 6 using 4-amino-N'-[3hydroxyethyl)sulfonyliminol-benzenesulfonamide (125mg, 0.31 mmole) and Libc102390 cyanuric chloride (28mg, 0.15mmole). The product was obtained as a colorless solid (105mg, 74%); m.p. >250°C dec; UV(DMSO): 298nm.

Example28 4,4'.dinitro-2,2'-diphenic acid 2,2'-diphenic acid (Aldrich, 50.0g, 206mmoles) was added portionwise to 500mL of 3:1 HNO31 H 2 S0 4 keeping the temperature less than 10 0 C. The clear solution was stirred in an ice bath for one hour, then carefully quenched over approximately 750g ice. A white solid is isolated by vacuum filtration which is a 2:1 ratio of the isomers shown. This mixture is purified by chromatographing down 1.5kg silica gel slurried in 8:2 isopropanol/ammonium hydroxide and eluted with the same solvent mixture. The product comes off the column first, practically running with the solvent front. The minor isomer comes off the column later (see P.R. 232-11). Product fractions are concentrated on the rotovap. The concentrated solution is acidified to pH1 with HCI, and filtered to afford 31.9g of 4,4'-dinitro-2,2'-diphenic acid. 300 Mhz NMR (DMSO d6): 67.55 (1H, d, 8.45 (1H, dd, J=2.5, 8.68 (1H, d, 13 (1H br CHN: Theory: C:48.60% H:2.77% N:8.10%.

Theory calculated with 0.75mole water. Found: C:48.68% H:2.72% N:7.42% Example 29 4,4'-diamino-2,2'-diphenic acid 4,4'-Dinitro-2,2'-diphenic acid (Example 28, 10.0g, 30.1mmol; P.R. 232-9) is suspended in water and titrated to pH7 with saturated NaHCO3 (NaOH may be used). 10% Pd/C Aldrich) is added and the mixture is reduced under 30psi H 2 for 1h. The catalyst is removed by filtration and the filtrate is concentrated to 20mL. Acetone (40mL) is added carefully to the concentrated solution, causing precipitation of the product as the disodium salt. It is collected by vacuum filtration and dried in vacuo to afford 9.2 g of 4,4'-diamino-2,2'-diphenic acid as the disodium salt. 300 MHz NMR (D 2 64.70 (NH 2 6.71 (1H, dd, J=2.4, 8.25); 6.79 (1H, d, 7.00 (1H, d, 1 25 J=8.25). CHN: Theory: C:49.64 H:3.72 N:8.27 (Calculated with 1.25mole water). Found: C:49.55 H:3.72 N: Example 4,4'-bis-(4.6-bis{3-[bis-(2-carbamoylethyl)-sulfamoyl]-phenylamino}-1,3,5-triazin-2-ylamino]-biphenyl-2,2'- Sdicarboxylic acid, disodium salt 30 A suspension of 4,4'-diaminobiphenic acid, disodium salt (Example 29, 66mg, 0.19mmol) and 2-chloro-4,6-di[3-aminophenyl-N,N-bis(2-carbamoylethyl)-sulfonyliminol-1,3,5-triazine (Example 6) (300mg, 0.40mmol) in sulfolane (10mL) and diisopropylethylamine (0.3mL) was warmed in a sealed thick-walled glass tube to 115°C. Warming was continued for 50h. Once the reaction was complete as determined by analytical HPLC (or TLC), the mixture was cooled to 20°C, and the product precipitated 35 by addition of propan-2-ol (70mL), washed on filter with cold ethanol (15mL), and dried under vacuum at 40°C. Two portions of a solution of crude product (about 0.3g) in deionised water (200mL) were purified separately by preparative RP-HPLC. Fractions containing the desired product were combined, organic solvents evaporated under vacuum (10mm Hg) at 30°C, and the product concentrated, desalted and isolated via octadecyl (C18) cartridge (900mg, obtained from Burdick Jackson). The cartridge was washed with water (20mL) and the product reextracted with methanol Libc/02390 Evaporation of the methanol gives 112mg of the desired product; m.p. >250'C dec; MS(ES) m/z: M- 2 840.2; MW 1724.4.

Example 31 3-aminophenyl-N,N-bis(2-methoxycarbonylethyl)sulfonylimine 3-Aminobenzenesulfonamide (3g, 17.4mmol) was dissolved in nitrobenzene (55mL) at 1000C.

Temperature was then reduced to 90*C and methyl acrylate (5mL, 55.5mmol) and Triton B (0.4mL, in MeOH) was added. The reaction vessel was equipped with a strong condensor and allowed to run for 20h. TLC showed the completeness of the reaction. A solution of HCI (30mL, 1M in diethyl ether) was added to the reaction mixture to produce a sticky oil. This was dissolved in 50% ethyl acetate/hexane and loaded onto a silica gel column. The desired product was eluted with hexane. The solvent was evaporated to give 3.9g of the desired product; (mlz): MH 343.2, mp 59-600C.

Example 32 3-aminophenyl-N,N-bis-(2-methylcarbamoylethyl)sulfonylimine To a solution of methylamine (10mL, 8.03M, 33% in ethanol) was added a solution of 3aminophenyl-N,N-bis(2-methoxycarbonyl-ethyl)sulfonylimine (Example 31, 0.5g, 1.45mmol) in ethanol with stirring at room temperature. After 24h, TLC showed the completeness of the reaction.

Water was added to the reaction and the compound was extracted with chloroform and dried over sodium sulfate (50mg). Yield is 300mg MS (ES) m/z 345.2 Example 33 3-[[3-(4-{3-[bis-(2-mtthylcarbamoylethyl)-sulfamoyl]-phenylamino}-6-chloro-1,3,5-triazin-2-ylamino)benzenesulfonyl]-(2-methylcarbamoylethyl)-amino]-N-methylpropionamide To a phosphate buffer (15mL, pH=7) at OoC, was added a solution of cyanuric chloride (550mg, 2.98mmol) in dioxane (4mL). To the milky mixture was added a solution of 3-aminophenyl-N,N-bis-(2methylcarbamoylethyl)sulfonylimine (Example 32, 2.0g, 5.84mmol) in dimethylformamide while pH was kept at 6.5-7.2 range by addition of 1N sodium hydroxide solution. After addition, the temperature was raised to 550C for 2h at which point HPLC analysis revealed the completion of the reaction. The mixture was diluted with water (100mL) saturated with sodium chloride and extracted with ethyl acetate (4x150mL). Concentration of combined extracts to a viscous oil (3.5g) which was taken up in acetonitrile (30mL) and left in a refrigerator overnight. The gummy precipitate was stirred 30 with ether (20mL) until all precipitates turned into a fine powder which was isolated through filtration.

An amber solid (1.8g, 78% yield) was obtained; m.p. 50'C dec.; MS (ES) m/z 795.7 Example 34 4,4'-bis-[[4,6-bis-[[3-[[bis-[3-(methylamino)-3-oxopropyl]amino]sulfonyl]phenyl]amino]-l ,3,5-triazin-2yl]amino]-[1,1'-biphenyl]-2,2'-disulfonic acid, disodium salt 35 A mixture of 3-[[3-(4-{3-[bis-(2-methylcarbamoy-ethyl)-sulfamoyl]-phenylamino}-6-chloro- [1,3,5]triazin-2-ylamino)-benzenesulfonyl]-(2-methylcarbamoyl-ethyl)-amino]-N-methyl-propionamide (Example 33, 200mg, 0.25mmol), 4,4'-diaminobiphenyl-2,2'-disulfonic acid, disodium salt (41mg, 0.11mmol) and diisopropylethylamine (50mL, 0.28mmol) in dimethylsulfoxide (4mL) was heated in a sealed tube at 110OC for 25h. The similar isolation procedure as above produced a white solid m.p. >2500C dec.; MS (ES) rn/z 930.5 (M 2 MW 1908.1.

Libc102390 22 Example 4,4'.bis-(4,6bis(3[bis(2methylcarbamoylethyl)sufamoyII-phelamilol-1 ,3,5-triazin-2-ylamino)biphenyl-2,2'-dicarboxylic acid, disodium salt To a solution of 3-[-41-bs(-ehlabmy-thl-ufmy]peyaio--hoo (1 ,3,5]triazin-2-ylamino)-belzelesulfonyl]-(2-methylcarbamoylethyl)-amiflo] N-methylpropionamide (Example 33, 200mg, 0.25mmol), 4 ,4'-diaminobiphenyl-2,2'-dicarboxylic ai, disodium salt (37mg, 0.l2mmol) in dimethylsulfoxide (4mL-) was added, a solution of (37mg, 0.l2mmol) in a phosphate buffer (3mL, pH=7). The mixture wis heated at 11000 for 24h. After cooling, water (lOOmL) was added, and the mixture was separated on a preparative HPLC column. Yield is 60mg of white solid; m.p. 5000 dec.; MS (ES) mlz 894.5 (M 2 MW 1836.0.

Example 36 4,4'bis-(4,6-bis{3[bis(2carbamoyethy)-sufamoyI]-penylamino)l ,3,5-triazin-2-ylamino)-biphenyl-2,2'dicarboxylic acid, dimethyl ester A mixture of 2-chloro-4,6-di[3-aminophenyl-N,N-bis(2-carbamoylethyl)-sulfonyliminoI-l triazine (Example 6, 200mg, 0.27mmol), 4,4'-diamino-2,2'-diphenic acid dimethyl ester 2 (38mg, 0.l13mmol) and dilsopropylethylamine (55mL, 0.31 mmol) was heated at 11000C for 30h. After cooling, isopropyl alcohol (50mL-) was added. The precipitate was filtered, washed with isopropyl alcohol. The dried solid (190mg) was purified by HPLC to give a white solid (53mg, 24% yield); m.p. >25000 dec.; MS (ES) mlz 854.8 (M+2H) 2 MW 1707.8.

Example 37 3-nitrophenylN,Nbis(2hydroxypropy)sufoflimifle To a stirred solution of diisopropanolamine (50.0g, 75mmol) in water (500mL) was added mnitrobenzenesulfonyl chloride (57.1g, 35mmol). The mixture was stirred vigorously for 25h at room .9 temperature. The resulting suspension was filtered, the solid washed with H 2 0 (2x400mL), dried and recrystallized from ethyl acetate/hexanes mixture to give the desired product as white crystals. The yield 65.8g m.p. 11 4-116*C; 1 H N MR (00013, 300 MHz) 68.67 J 5.1 Hz, 1 8.44 1 H), 8.15 J 6.0Hz, 1 7.77 (in, 1IH) 4.78 2H), 4.18 (in, 2H), 3.10 J 5.9Hz, 4H) 1. 16 (dd, 6H); MS (ES) m/z 319.5 Example 38 3-aminopheny-N,N-bis(2-hydroxypropy)-sufoflimifle A solution of 3-nitrophenyl-N,N-bis(2-hydroxypropyl)sulfonylimine (Example 37, *,***31.4mmol) in methanol (lO0mL) was hydrogenated over the palladium catalyst (10% Pd/C, 1g) for 1 .5h. The resulting mixture was filtered through a pad of Celite, the solvent was removed and the resulting viscous oil was purified by column chromatography on Silica Gel, elution by ethyl 35 acetate/hexane 4:1 mixture. The desired product was obtained as light-yellow crystals. The yield 8.2g *fee NMR (00013, 300MHz) 6 7.28 1 7.13 (in, 1 7.09 (in, 1IH), 6.86 1 H) 4.20 (in, 1 H), 4.13 (dd, 1 3.37 (dd, 1 3.02 2H), 2.80 (dd, 1IH); 1. 16 (dd, 6H); MS (ES) m/z 289.5 Example 39 2-chloro-4,6-bis{3-[bis(2-hydroxy-propyI)4sufamoyl]Fphenylamino-1 The title compound was prepared according to the procedure of Example 33, using cyanuric chloride (160mg, 0.87mmol) 3-aminophenyl-N,N-bi(2-hydroxypropyl)sulfonylimfine (700mg, 2.Ommol), Libc/02390 dioxane (3mL), phosphate buffer (l0mL-, 0.3M, pH-7), N,N-dimethylformamide (5mL) and 1N NaOH.

The product was precipitated from the cooled reaction mixture with water as a viscous amber-yellow oil, centrifuged,washed with water and dried on the high vacuum to give 640mg of the chlorotriazine. m. p. >25000 dec; MS (ES) mlz 687.7, 689.7 Example 4 4 1.bis(4,6bis-3[bis.(2hydroxypropy)-sulfamoylF-phelylamilo}-,3,5,-triazin-2-ylamino)biphenyl-2,2'disulfonic acid, disodium salt A mixture of 2-hoo46bs[-bi-2hdx-my)slfmy (Example 39, 300mg, 0.44mmol), 4,4'-diaminobiphenyl-2,2'-disulfonic acid, disodium salt (77mg, 0.2mmol), 2mL of DMSO, 2mL of the phosphate buffer (1N, pl-i) and 0.25mL of 1N NaGH was exposed to a microwave heating (PROLABO unit, monomode regimen) at 11000C for 2h. After cooling the reaction mixture without workup was submitted to preparative HPLC in water/acetonitrile system on YMC Prodigy 018 polymer column. The product was obtained as colorless solid (170mg, 50.2%); m. p. >2500C dec; MS (ES) mlz 822.3 (M-2H) 2 MW 1691.9 (MW+2Na) Example 41 4,4'bis.(4,6-bis.{3.rbis-(2-hydroxypropyl).sulfamoyl]-phenylaminol ,3,5.triazin-2.ylamiflo)bipheflyl-2,2'dicarboxylic acid, disodium salt A mixture of 2-hoo46bs(-bi-2hdoymy)slfmy (Example 39, 178mg, 0.25mmol), 4,4'-diamino-2,2'-diphenic acid, dihydrochloride (35mg, 0.11lmmol), 3mL of DMSO, 0.5mL of the phosphate buffer (i N, pH 7) and 0.5mL of 1IN NaOH was heated in the oven in a sealed tube for 18h, then without a workup submitted for the prep. HPLC to give of the desired product. m. p. >2500C dec; MS (ES) mlz 788.1 (M+2H) 2 MW 1619.8 a(MW+2Na) 0. 0 Example 42 0:9.0: 25 2-chloro.4,6bis-{3[bis-carbamloylmethyli1 C. *6 The title compound was prepared according to the procedure of Example 33, using cyanuric :980 chloride (0.42g, 2.27mmoI), 3-mnpey-,-i-abmymty-ufnlmn (CL 55675, 1 .43g, dioxane (2mL), phosphate buffer (l0mL-, 0.3M, pH-7), N,N-dimethylformamide (4mL-) and NaOH. The reaction mixture was heterogeneous during the whole procedure. The formed pink a..

30 precipitate was centrifuged, washed several times with water, redissolved in a minimal amount of DMF and re-precipitated by water. The final separation and drying gave 0.7g (45.1 of the desired product; m. p. >25000 dec; MS (ES) mlz 683.7, 685.7 Ilto.Example 43 4s,4.bis{(4,6bis13(biscarbamoylmethyl.1 .sulfamoyl-phenylamino]-l ,3,5-triazin.2-ylamino}-biphelyl-2, 2 *o 35 disulfonic acid, disodium salt toocs The title compound was prepared according to the procedure of Example 40, using 2-chloro- 4 ,6-bis-{3-bis-carbamoylmethyl-1 -sulfamoyll-phenylamino}-l ,3,5-triazine (Example 42, 500mg, 0.73mmol), 4,4'-diaminobiphenyl-2,2'-disulfonic acid, disodium salt (1 14mg, 0.29mmol), 2.5mL of DMSO, 2.5mL of the phosphate buffer (iN, pH-7) and 1.OmL of 1N NaCH. The microwave heating (PROLABO unit, monomode regimen) was continued for Ih at 10500. The resulting mixture was Libc[02390 submitted for the prep. HPLC, which gave 50mg of the product as a pink solid; m. p. >250°C dec; MS (ES) m/z 818.2 (M-2H) 2 MW 1683.7 (MW+2Na) Example 44 4',4-bis-{4,6-bis-[3-(bis-carbamoylmethyl-1-sulfamoyl)-phenylamino]-1,3,5-triazin-2-ylamino}-biphenyl-2.2'dicarboxylic acid, disodium salt The title compound was prepared according to the procedure of Example 40 using 2-chloro-4,6bis-{3-[bis-carbamoylmethyl-1-sulfamoyl]-phenylamino}-l,3,5-triazine (Example 42, 500mg, 0.73mmol), 4,4'-diamino-2,2'-diphenic acid, dihydrochloride (100mg, 0.29mmol), 2.5mL of DMSO, of the phosphate buffer (1N, pH7) and 1.0mL of 1N NaOH. The microwave heating (PROLABO unit, monomode regimen) was continued for 1h at 105°C The resulting mixture was submitted for the prep. HPLC, which gave 20mg of the product as a pink solid; m. p. >250°C dec; MS (ES) m/z 784.0 (M+2H) 2 MW 1619.8 (MW+2Na).

Example 1-(3-nitrobenzenesulfonyl)piperidine-3,5-dicarboxylic acid A solution of 3,5-pyridinedicarboxylic acid (3.34 g, 20mmol) in 50mL of water and 10mL of conc. ammonium hydroxide was hydrogenated in Parr apparatus at 22psi over 1g of ruthenium catalyst ruthenium on alumina powder) for 48h. After filtration of the catalyst and solvent removing the crude 3,5-piperidinedicarboxylic acid was dissolved in 60mL of 1N NaOH and treated with 6.6g (30mmol) of m-nitrobenzenesulfochloride. The reaction mixture was stirred at room temperature and pH maintained at 9.5 by addition of 1N NaOH until it stopped to drop down. The reaction mixture was stirred for an additional 1.5h, the formed precipitate was filtered, and the filtrate was acidified by conc. HCI to pH2-3. The formed white precipitate was filtered, washed with water and dried in high vacuum overnight to give 5.3g of the desired product. 1H NMR (DMSO-d6, 300MHz) 8 8.55 1H), 8.40(s, 1H), 8.15 1H), 7.95 1H) 3.14 4H), 1.65 2H), 1.45 (m, 25 2H); MS (ES) m/z 359 (M+H) Example 46 1-(3-nitrobenzenesulfonyl)piperidine-3,5-dicarboxylic acid dimethyl ester A suspension of 1-(3-nitro-benzenesulfonyl)piperidine-3,5-dicarboxylic acid (Example 45, 3.6g, 10mmol) in THF (200mL) was treated at 0°C by the excess of diazomethane and stirred at that o 30 temperature until the suspension turned into clear bright-yellow solution. The reaction mixture was warmed to the room temperature, filtered, evaporated and purified by column chromatography on Silica Gel, elution by chloroform, to give 2.5g of the desired products colorless crystals. 'H *ass NMR (CDCl 3 ,300MHz) 8 8.65 1H), 8.49(d, 1H), 8.11 1H), 7.81 1H) 3.73 6H), 3.40(m, 4H), 2.95 2H), 2.05 2H); MS (ES) m/z 387.2 35 Example 47 1.(3.nitrobenzenesulfonyl)piperidine-3,5-dicarboxylic acid diamide A solution of 1-(3-nitro-benzenesulfonyl)piperidine-3,5-dicarboxylic acid dimethyl ester (Example 46, 2.5g, 6.5mmol) in 200mL of methanol, saturated by ammonia at 0°C, was sealed and left at the room temperature for 7 days. The resulting yellow solution was concentraed to the volume Libc02390 100mL, the formed precipitate filtered, washed with methanol and dried to give 1.8g of the product as light-creamy crystals. MS (ES) m/z 357.1 Example 48 1-(3-amino-benzenesufonyl)piperidine-3,5-dicarboxylic acid A solution of 1-(3-nitro-benzenesulfonyl)piperidine-3,5dicarboxylic acid (Example 45, 4.2mmol) in 150mL of methanol was hydrogenated in Parr apparatus at 25psi over 0.15g of the palladium catalyst (10% palladium on carbon) for 1h. Filtration of the catalyst and solvent removing gave 1.8g of the desired product as a white solid. MS (ES) m/z 329.1 Example 49 1-(3-amino-benzenesulfonyl)piperidine-3,5dicarboxylic acid diamide A solution of 1-(3-nitro-benzenesulfonyl)piperidine-3,5-dicarboxylic acid diamide (Example 47, 4.2mmol) in 300mL of methanol was hydrogenated in Parr apparatus at 25psi over 0.lg of the palladium catalyst (10% palladium on carbon) for 1.5h. Filtration of the catalyst and solvent removing gave 1.26g of the desired product as a light-gray solid. MS (ES) m/z 327.1 Example 2-chloro-4,6-bis-[3-(3,5-dicarbamoyl-piperidine- -sulfonyl)-phenylamino]-1,3,5-triazine A solution of cyanuric chloride (280mg, 1.5mmol) in dioxane (5mL) was added with stirring to phosphate buffer (10mL, 0.3M, pH7) at 0-20C. To the resulting suspension was added dropwise a solution of 1-(3-amino-benzenesulfonyl)piperidine-3,5dicarboxylic acid diamide (Example 49, 20 3.lmmol) in N,N-dimethylformamide (5mL) and the pH maintained at 6.5-7.2 by addition of 1N NaOH.

After completion of the addition, the reaction mixture was warmed up to 50-550C and stirred at that temperature for 2h (until HPLC showed the completion of the reaction), then cooled to the room temperature, diluted with water, the product precipitate filtered, washed with cold water acetone (5mL) and dried under vacuum at 200C to give 845mg of the desired product; m.p.

25 >2500C dec; MS (ES) m/z 765.0 MW 764.2 Example 51 2-chloro-4,6-bis-[3(3,5dicarboxylpiperidine-1-sulfonyl)-phenylamino]-1,3,5-triazine The title compound was prepared according to the procedure of Example using cyanuric chloride (280mg, 1.5mmol); 1-(3-amino-benzenesulfonyl)piperidine-3,5dicarboxylic acid (Example 48, 1.0g, 3.lmmol), dioxane (3mL), phosphate buffer (10mL, 0.3M, pH7), N,N-dimethylformamide and 1N NaOH. After completion of the reaction the mixture was cooled to the room temperature, diluted with water, acidified by HCI to pH5, the product precipitate filtered, washed with cold water (1OmL), and dried under vacuum at 200C to give 845mg of the desired product; m.p. >2500C dec; MS (ES) m/z 769.0 MW 767.8.

Example 52 ,2.ethenediyl)bis[5-[[4.6-bis[-3-[[3,5-bis(aminocarbonyl)-l -piperedinyl]sulfonyl]phenyl]amino]- 1,3,5-triazin-2-yl]amino]benzenesulfonic acid], disodium salt A suspension of 4,4'-diaminostilbene-2,2'-disulfonic acid, disodium salt (0.41g, 0.lmmol) and 2chloro-4,6-bis-[3-(3,5-dicarbamoyl-piperidine-1 -sulfonyl)phenylamino]-l ,3,5-triazine (Example (1.62g, 0.2mmol) in sulfolane (3mL) and dimethylformamide (5mL) in the presence of Libcl02390 dilsopropylethylamine (0.08mL) was warmed in a sealed thick-walled glass tube to 1150C. Warming was continued for 40h. Once the reaction was complete as determined by analytical HPLC (or TLC), the mixture was cooled to 300C, 1N NaOH (0.5mL) was added and the product precipitated by the addition of propan-2-ol (50mL). The precipitate was filtered, washed with acetone (30mL) and redissolved in a minimum volume of water and re-precipitated by the addition of ethanol. The precipitate after filtration was washed with ether (30mL) and dried under vacuum at 500C. Yield 1.59g MS(ES) m/z 911.7 (M-2H) 2 MW 1869.9(2Na).

Example 53 4',4-bis-{4,6-bis-[3-(3,5-dicarbamoylpiperidine- -sulfonyl)-phenylamino]-l ,3,5-triazin-2-ylamino}-biphenyl- 2,2'-disulfonic acid, disodium salt The title compound was prepared according to the procedure of Example using 4,4'diaminobiphenyl-2,2'-disulfonic acid, disodium salt (40mg, 0.lmmol) and 2-chloro-4,6-bis-[3-(3,5dicarbamoylpiperidine-1-sulfonyl)-phenylamino]-1,3,5-triazine (Example 50) (176mg, 0.23mmol) in DMSO sulfolane v/v) and diisopropylethylamine (0.08mL). The product was obtained as a creamy solid (153mg, m.p. >2500C dec; MS(ES) (mlz) 898.3 (M-2H) 2 MW 1843.9 (M+2Na).

Example 54 "'-[(2,2'-disulfo[l ,I'-biphenyl]-4,4'-diyl)-bis[imino- ,3,5-triazine-6,2,4-triylbis(imino- 3 ,1acid The title compound was prepared according to the procedure of Example using 4,4'- 20 diaminobiphenyl-2,2-disullonic acid, disodium salt (270mg, 0.68mmol) and 2-chloro-4,6-bis-[3-(3,5dicarboxylpiperidine--sulfonyl)-phenylamino]-1, 3 ,5-triazine (Example 51) (1.1g, 1.43mmol) in DMSO-sulfolane v/v) and diisopropylethylamine (1.2mL). The product after precipitation by the addition of propan-2-ol was filtered, washed with acetone (30mL), dried, redissolved in a minimum volume of water and re-precipitated by acidifying (HCI) to pH1-2. The desired product was obtained as a creamy solid (1.0g, m.p. >250'C dec; MS(ES) 902.2 (M-2H) 2 MW 1807.8.

Example 3-nitrophenyl-N,N-bis(2-hydroxyethyl)sulfonylimine Diethanolamine (Aldrich; 25.0g, .238mol) was dissolved in 55mL 9:1 methylene chloride/THF and chilled to 50C. A solution of 3-nitrobenzenesulfonyl chloride (Aldrich; 25.0g, .113mol) in THF was added over 10 minutes. Ice bath was removed; reaction stirred for an additional 30 minutes.

Then the solvents were stripped off and the residue was partitioned between water (100mL) and ethyl acetate (150mL). The aqueous layer was ev-racted with three 25mL portions of ethyl acetate. The combined organic layer was dried over MgSO4, filtered through a pad of silica and concentrated in vacuo. The product crystallizes out during this evaporation and is filtered, giving 17.4g of white needles. m.p. 99-100*C. MS(ES) m/z 291.2 Example 56 3-aminophenyl-N,N-bis(2-hydroxyethyl)sulfonylimine The nitro compound (Example 55, 17.2g, 59.3mmol) and 1.72g of 10% Pd/C were suspended in 170mL ethanol. The mixture was reduced under 40psi hydrogen for 2h. The catalyst was filtered off Libc/02390 and the clear colorless filtrate was evaporated to dryness, giving 15.4g (98% yield) of the corresponding amino compound. MS(ES) m/z 261.0 (M+H) 1 Example 57 4 4 '-bis-(4,6-bis-{3-[bis-(2-hydroxyethyl)-sulfamoyl]-phenylamino}-1,3,5-triazin-2.ylamino)biphenyl-2,2'dicarboxylic acid, disodium salt A solution of trichlorotriazine (1.55 g; 8.4mmol) in 20mL dioxane was added to 60mL of pH7 buffer at 0°C. To the resulting fine suspension was added dropwise a solution of 4,4'-diamino-2,2'diphenic acid 232-10; 1.55g, 4.0mmol) in 10mL water, keeping the pH6.5-7.1 with 1N NaOH.

Once the addition is complete and the pH has stabilized, the reaction mixture is allowed to warm to ambient temperature. A solution of the diethanolsulfonamide (2.6g, 10mmol) in 20mL dioxane and solid sodium bicarbonate (0.84g, 10mmol) are added and the resulting mixture is heated at reflux (92 0 C) for two days. The clear yellow solution is cooled and the dioxane is stripped off. The product precipitates out of aqueous solution and is collected by vacuum filtration giving 3.16g of the desired compound, approximately 65% pure by HPLC as a white amorphous solid, m.p. >250°C dec; MS(ES) 732.5 (M+2H) 2 MW 1507.6 (M+2Na).

Example 58 3.aminophenyl-N,N-bis(3-hydroxypropyl)sulfonylimine A solution of 3-aminophenyl-N,N-bis(2-methoxycarbonyl-ethyl)sulfonylimine (Example 31, 150mg, 0.4mmol) in THF (15mL) was added to a solution of LiAIH4 (3mL, 1M in THF) with stirring under niteogen at 0°C. After 45 minutes, TLC shows the completeness of the reaction. A solution of °ammonium chloridr (15mL, 1M) was added to the solution and a white precipitate formed and solution :.turned yellow. The yellowish solution was decanted and the white precipitate was washed with ethyl acetate. The organics were dried over sodium sulfate and rotary evaporated to give a desired product as a yellow oil. Yield is 110mg MH 289.2.

25 Example 59

S

4 4 '-bis[4,6-di[3-aminophenylN,N-bis(3-hydroxypropyl)-sulfonyliminol-1,3,5-triazin-2-ylamino]-biphenyl- 2,2'-disulfonic acid, disodium salt The title compound was prepared using the general conditions of Example 1, where cyanuric chloride (92mg, 0.50mmol) was reacted with 4,4'-diamlnobiphenyl-2,2'-disulfonic acid, disodium salt (93mg, 0.24mmol) for 1h, and then with 3-aminophenyl-N,N-bis(3-hydroxypropyl)sulfonylimine (708mg, 2.5mmol) (Example 58). Once the reaction was complete as determined by analytical HPLC (or TLC), the mixture was cooled to 20°C, and the product precipitated by addition of propan-2-ol washed on filter with cold propan-2-ol (15mL), and dried under vacuum at 40 0 C. Two portions of a solution of crude product (about 0.35g) in deionised water (200mL) were purified separately by preparative RP-HPLC. Fractions containing the desired product were combined, organic solvents evaporated under vacuum (10mm Hg) at 300C, and the product concentrated, desalted and isolated via octadecyl (C18) cartridge (900mg, obtained from Burdick Jackson). The cartridge was washed with water (40mL) and the product reextracted with methanol (60mL). Evaporation of the methanol gives 162mg of the desired product; m.p. >300°C dec; MS(ES)

M-

2 822.4; MW 1690.8.

Libc/02390 28 Effects of the compounds in viral yield reduction assays.

The compounds were tested for biological activity using viral yield reduction assays. Host cells (indicated as HF for human foreskin fibroblasts and as vero, the commonly used African green monkey cell line, in Tables 1 and la) were plated at 4x10 4 cells/well in cell growth medium with 2% fetal bovine serum or 2% calf serum in 96-well plates (0.lmL/well) and incubated overnight. The compounds were added to individual wells to give final concentrations ranging from 50 to 0.1 g/mL in two-fold concentration steps. After Ih, virus was added, and the incubation was continued for 4 days with RSV or CMV and for 2 days with HSV. At the end of the experiment, the amount of virus growth was quantitated and the yield reduction at various concentrations of compound was calculated. This allows estimation of the concentration of compound giving 50% reduction in virus growth Tables 1 and la (a summary of biological activity) summarizes the activity of the compounds in viral yield reduction assays.

Effects of the compounds in viral plaque reduction assays.

The compounds were tested for the ability to inhibit viral plaque formation. Host cells, as indicated in Tables 1, la and 2, are plated at 106 cells/well in 6-well plates and incubated overnight.

One of two formats was then followed. In the normal (pretreatment) format, the cell growth medium was replaced with l mL of medium containing compound at a given concentration. Virus (-100 plaqueforming units in 40ptL) was added after I h. After a further hour, the medium was replaced with 4mL of medium containing compound at the concentration being tested. The cells were then incubated for a S 20 time sufficient to allow the formation of viral plaques. Occasionally, compound was added only after the infection had begun (posttreatment). In this case, the growth medium was replaced with 1 mL of medium containing approximately 100 plaque-forming units. After lh, this was replaced with 2mL of growth medium. After a further period specified in Table 2, a further 2mL containing compound was added. In either case, the number of plaques in a series of wells containing a range of compound ~25 concentrations was compared to the number in a well with no compound, and the IC50so was determined by graphing the data and identifying the compound concentration giving a 50% reduction in viral plaque number.

Tables 1, la and 2 summarize antiviral activity in plaque assays. It can be seen that the title *e compounds of Examples 1 and 4 are active against all RSV strains tested at approximately 0.1- 0.7 1 g/mL. Activity can also be seen with certain members of other virus families (in the range 3tg/mL). These results confirm and extend the data from yield reduction assays indicating that these compounds are potent and specific inhibitors of RSV.

Libc/02390 29 Table I In vitro activity of the compounds against Respiratory Syncitial Virus, Cytomegalovirus, and Herpes Simplex Virus Compound of ICso pg/mL IN GROWTH INHIBITION ICso Iig/mL IN PLAQUE ASSAY Example RSV/HF RSVNERO CMV/HF HSV/HF HSVNERO RSV/VERO CMV/HF HSVNERO 11 10 10 9 <3 0.4 0.3 <3 0.7 6 >50 >50 1 0.3 0.5 0.3 7 8 4 0.1 0.1 35 20 35 0.1 >30 0.4 8 7 0.7 1 0.8 8 >50 >50 12 13 8 8 20 10 7 >8.3 2 1 16 5 -8.3 3 3 17 >50 >50 19 2 3 12 30 0.5 22 -0.15 0.25 4 2 23 8 24 Table 1. Fifty percent inhibitory concentrations for growth inhibition (yield reduction) and for plaque formation were determined as described in the text. The virus strains used were RSV (A2), CMV (Ad169), HSYI (Patton). The host cells used were human foreskin fibroblasts (HF) and vero cells.

Table la In vitro activity of the compounds against Respiratory Syncytial Virus, Cytomegalovirus and Herpes 99 r 9 Fifty percent inhibitory concentration for growth inhibition (yield reduction) and for plaque formation were determined as deescribed in the text. The virus strains used were RSV CMV (Ad169), and HSV1 (Patton). The host cells used were human foreskin fibroblasts (HF) and vero cells.

N. no effect on the appearance of the alcohol fixed and crystal violet stained cell monolayer at the indicated compound concentration. L, lightening of the intensity of staining (compared to controls) Libc/02390 of the alcohol fixed and crystal violet-stained cell monolayer at the indicated compound concentration but not at half that concentration.

Table 2.

Effect of Examples 1 and 4 on viral plague formation Virus Strain ICo0 (ig/mL) (Cell) Example 1 Example 4 Pre Post (hours) Pre Post (hours) 2 24 2

RSV(VERO)

A2 0.3 0.3 0.4 0.1 0.28 LONG 0.2 0.21 3A 0.5 0.15 9320 0.2 0.18 2B 0.1 0.08 RSS2 0.20 18736 0.06 RSV (HF A2 0.4 <3

RSV(SKNSH)

A2 0.4 RSV (MDBK) A2 0.4 HPIV3 (VERO) WASH FLU (MDCK) AI/FM/1/47 A2/HK/8/68 A2/JPN/305/57 A/Texas/36/91 A/Beij/32/92 B/Pana/45/90 eHCMV (HF) Ad169 15

HSYI(VERO)

Patton HSV2(VERO) 333 3 12 MS S In the pretreatment protocol, confluent cells in 6-well dishes were treated with various concentrations of compound for 1h. Virus (~1OOpfu) was added in a small volume and plaques were counted after incubation (RSV, CMV, and HPIV3 (human parainfluenza 45 virus type 3, strain Washington), 5-8 days; influenza and herpes simplex virus, 2 days). For posttreatment, cells were first infected for 1h and the medium (containing unabsorbed virus) was replaced. Medium containing compound at various concentrations was added at 2h or 24h after the end of the infection period. Cells used included vero (African green monkey cells), HF (human foreskin fibroblast cells), SKNSH (human neuroblastoma cells), MDBK (Madin-Darby bovine kidney cells), and MDCK (Madin-Darby canine kidney cells).

Mechanism of action.

The time in the virus life-cycle when Example 1 exerts its effect was determined to increase understanding of the mechanism of action. This was accomplished by adding and removing the compound at various times relative to infection and assessing the effect on viral protein synthesis.

Vero cells were infected on ice for 1h, unadsorbed virus was removed, and then the infected cells Libc/02390 were incubated at 37°C for 18h (virus adsorption will occur on ice, but fusion will not proceed until the temperature is raised to 18°C or above). After the 18h incubation at 370C (when the bulk of ongoing protein synthesis was directed by viral RNAs), 3 5 S-methionine was added for 2h, the cells were lysed, and the labeled proteins were analyzed on SDS-PAGE gels. The viral N, P, and M proteins were then evident. If the compound of Example 1 was added at 10 Og/mL immediately after virus adsorption and before raising the incubation temperature to 37°C, viral protein synthesis was fully prevented.

However, when the compound of Example 1 was added even after only 5 minutes incubation at 370C, viral protein production was unimpeded. Therefore, the process inhibited was the fusion process or some closely succeeding step which is required for later viral gene expression.

It is known that RSV-infected cells in close contact will fuse to form syncytia and that this process is dependent only on the viral F, SH, and G proteins (Heminway BR, Yu Y, Tanaka Y, Perrine KG, Gustafson E, Bernstein JM, Galinski MS. 1994. Analysis of respiratory syncytial virus F, G, and SH proteins in cell fusion. Virology 200;801-805). Syncytium formation should not involve viral functions, such as uncoating or transcription, which normally succeed it and which are required for later viral gene expression.

To determine if the compounds of this invention inhibit fusion or a succeeding step, they were tested for the ability to prevent syncytium formation in cells previously infected with RSV. This was done by infecting vero cells at a multiplicity of infection of 3 for 1 h, then replacing the original medium with fresh medium. After 6h incubation, compound was added. After 24 and 48h, the infected cells 20 were examined under the microscope to determine the extent of syncytium formation. Differences in the extent of syncytium formation due to the presence of compound were easily recognized and the compounds were easily ranked in terms of potency in this assay. The actual iCsos as judged by this method are approximate. They were 0.3 and 0.05 g/mL for compounds 16 and 17, respectively. The ability of these compounds to inhibit syncytium formation demonstrates that they act at the fusion step 25 of the virus life-cycle.

Compounds of Examples 1 and 4 prevent RSV growth in vivo.

.RSV infection of cotton rats followed by drug delivery by small particle aerosol (SPA) was used to assess a compound's in vivo anti-RSV potential. (Gilbert BE, Wyde PR, Wilson SZ, and Meyerson LR. 1993. SP-303 small particle aerosol treatment of influenza A virus infection in mice and 30 respiratory syncytial virus infection in cotton rats. Antiviral Research 21 ;37-45.) The compounds of the present invention were tested in this system to determine if they can inhibit growth of RSV in cotton rats. In experiment 1 (see Table animals were infected intranasally with RSV A2 (day which results in a respiratory infection. Ribavirin was used at 60mg/mL in water in the aerosolizing chamber and was administered for 2h twice a day on days 1, 2, and 3 after infection. Compound 16 was used at 4.5mg/mL in 25mM NaHCO3 in the aerosolizing chamber and was administered for 4h before infection and for 8h on each of days 1, 2, and 3. The placebo was water used according to the same regimen as compound 16. The animals were sacrificed on day 4, the lungs were lavaged, and the titers of RSV in the lavages determined. In experiment 2, infected untreated animals and animals exposed to an aerosol of 25mM NaHCO3 served as controls. The compounds of Examples 1 and 4 were made up at 5mg/mL in 25mM NaHCO3. One group was treated Libc/02390 32 with the compound of Example 1 for 4h before infection on day 0 and for 8h on each of days 1, 2, and 3 (prophylactic regimen). One group was treated with the compound of Example 1 for 8h only on each of days 1, 2, and 3 (therapeutic regimen). A final group was treated with the compound of Example 4 for 8h on each of days 1, 2, and 3 (therapeutic regimen). On day 4, all animals were sacrificed, the lungs were lavaged, and the titers of RSV in the lavages were determined. In experiment 3, the placebo was water delivered intranasally once a day. The compound of Example 4 was tested by intranasal delivery by dosing before inoculation and 1 day thereafter (days 0 and by dosing before inoculation and on days 1-3, and by dosing only after inoculation on days 1-3.The compound was also tested by dosing intraperitoneally. On day 4, all animals were sacrificed, their lungs were lavaged, and the titers of RSV in the lavages were determined. This was done by determining the dilution end-point which produces cytopathic effect (CPE) in vero cells and also by plaque assay on vero cells. The lungs were a~o homogenized and the titers of virus in the lungs were determined by centrifuging out particulate material (1000xg, 10 minutes) and using dilutions of the supernatants in plaque assays. It can be seen that the compounds of Examples 1 and 4 inhibit RSV in vivo.

Table 3.

Efficacv of Examoles and 4 aainst Respiratory ncitial Virus in Coton Rats

EXPT

1 i SD Fold .r r r r r r r

EXPT

1 2 3 Treatment Regimen 20 aerosol 2x2h/d dl,2,3 Ribavirin Aerosol 60mg/mL2x2h/d dl,2,3 Example 1 Aerosol 4.5mg/mL 4h dO 8h dl,2,3 Jntreated Placebo Aerosol 4h d08h d1,2,3 Example 1 Aerosol 5mg/mL 4h d08h d1,2,3 Example 1 Aerosol 5mg/mL8hd1,2,3 Example 4 Aerosol 5mg/mL8hd1,2,3 H20 i.n. d0,1,2,3 Example 4 i.n. dO 15mg/kg dl 30mg/k Example4 i.n. dO 15mg/kg d,2,330mg/kg Example 4 i.n. 30mg/kg d1,2,3 Example 4 i.p. 30mg/kg d1,2,3 1.

Sample/Assay Lavage/CPE Lavage/CPE Lavage/CPE Lavage/CPE Lavage/CPE Lavage/CPE Lavage/CPE Lavage/CPE Lavage/CPE Lavage/Plaque Lung/Plaque Lavage/CPE Lavage/Plaque Lung/Plaque Lavage/CPE Lavage/Plaque Lung/Plaque Lavage/CPE L avage/Plaque Lung/Plaque Lavage/CPE Lavage/Plaque Lung/Plague Mean titer log10/g lung 4.1 3.3 2.8 3.68 3.43 1.73 3.43 2.05 4.18 4.29 4.55 1.66 2.26 2.32 2.05 2.25 2.25 1.67 3.09 3.37 4.18 4.1 3.92

SD

0.27 0 0.35 0.48 0.25 1.15 0.25 0.5 0.25 0.13 0.24 0.25 0.11 0.23 0.87 0 0 0.48 0.65 1.09 0.25 0.22 0.97 Fold 0.8 1.3 0.25 1.95 0.25 1.63 2.52 2.03 2.23 2.13 2.04 2.3 2.51 1.2 1.18 0 0.09 0.26

I

I

Libc/02390 33 The claims defining the invention are as follows: 1. A compound having the structure: X N, BAB' N X NUN A NUN B' B'

U

C,

C,

Q

S

wherein: A is eeeeC,

I

C,

Sor C' is-

SO

3 H, OSO 3 H, OH, or COOH; B' is NH, NR 1 or O; R 1 is H, (Cl-C 6 )lower alkyl, straight or branched, wherein the carbon atoms may be optionally substituted with CI, Br, F, OH or CN; X is CI, F or the /z -N YZ m moiety 'w U' is SO 2 -CO, or W' is: z

Z

-N N m or z Y is (CH2)n; n is 0 to 6; m is 0 to 2; Z is H, CH 3 CF3, CH 2 -(halogen), where halogen is CI, Br, F or I, CH20H, -COOH, COO(Ci-C6)lower alkyl straight or branched, -CONR 2

R

2 2

R

CN or OH and R 2 in each occurrence, is independently H or (C1-C6)lower alkyl; and the pharmaceutically acceptable salts and esters thereof.

Libc/02390

Claims (47)

1. 2. A compound of claiml1wherein Ais the moiety: C is S031-; B' is -NH; Wis the moiety: ;Y is CH 2 Z CH 2 OH; and U, X, Y, n, and m are as defined in claim 1; or a pharmaceutically acceptable salt or ester thereof. C. C.
2. 3. A compound of claim 1 wherein A is the moiety: C1 I-Z is -SO 3 H; B3' is NH; W' is the moiety: Y is CH 2 CH2; Z CONH- 2 and X, Y, n, and mn are as defined in claim 1; or a pharmaceutically acceptable salt or ester thereof.
3. 4. A compound of claim 1 wherein A is the moiety: C' is S0 3 B' is NH; W' is the moiety: Y is CH 2 CH2; Z CONH-2; and X, Y, n, and mn are as defined in claim 1; or a pharmaceutically acceptable salt or ester thereof.
4. 5. A compound as claimed in any one of claims 1 to 4, which is a disodium salt as pharmaceutically acceptable salt.
5. 6. 4,4'-bis[4,6-di[3-aminophenyl-N ,N-bis(2-carbamoylethyl)sulfonylimino]-l ,3,5-triazin-2- ylamino]-stilbene-2,2'-disulfolic acid or a pharmaceutically acceptable salt thereof.
6. 7. 4,4'-bis[4,6-di[3-aminophenyl-N ,N-bis(2-carbamoylethyl)sulfonylimifl-l ,3,5-triazin-2- ylamino]-biphenyl-2,2'-disulfonic acid or a pharmaceutically acceptable salt thereof.
7. 8. 4,4'-bis[4,6-di[3-aminophenyl-N ,N-bis(2-hydroxyethyl)sulfonylimiflo]-l ,3,5-triazin-2- ylamino]-stilbene-2,2'-disulfonic acid or a pharmaceutically acceptable salt thereof.
8. 9. 4-[4,6-di[3-aminophenyl-N ,N-bis(2-carbamoylethyl)sulfonylimino]-l ,3,5-triazin-2-ylaminol- 4'-[4-chloro-6-[3-aminophenyl-N ,N-bis(2-carbamoylethyl)sulfonylimino]-1 ,3,5-triazin-2-ylaminol- stilbene-2,2'-disulfonic acid or a pharmaceutically acceptable salt thereof. 4,4-i[-hlr--3amnpey ,N-bis(2-carbamoylethyl)LulfonyliminoI-l 2-ylaminoj-stilbene-2,2'-disulfonic acid or a pharmaceutically acceptable salt thereof.
9. 11. 4-[4,6-di[3-aminophenyl-N ,N-bis(2-carbamoyethyl)ulfoflylimino]-1 ,3,5-triazin-2-ylamino]- 4'-[4-[3-aminophenyl-N N-bis(2-ccrbamoylethyl)sulfonylimino]-6[3aminophenylN( 2 carbamoylethyl)N'-(3-propionic acid)]- 1,3,5-triazin-2-yl amino]-tilbene-2,2'-d isu fonic acid or a pharmaceutically acceptable salt thereof.
10. 12. 4-[4,6-di[3-aminophenyl-N ,N-bis(2-carbamoylethyl)sulfonylimino]-l1,3,5-triazin-2-ylamino]- 4'-[4-[3-aminophenyl-N, N-bis(2-carbamoylethyl)sulfonylimino]6[ 3 aminophenyl-N,N-bis(3-propionic Libc/02390 acid)]-1, 1 3,5-triazin-2-ylami no]-stilbele-2,2'-d isulfoflic acid or a pharmaceutically acceptable salt thereof.
11. 13. 4 ,4'-bis[4,6-di[3-aminophelyl-N ,N-bis(2-carbamoyethyl)sulfonylimilo]-1 ,3,5-triazin-2- ylamino]-bibenzyl-2,2'-disulfoflic acid or a pharmaceutically acceptable salt thereof.
12. 14. 4,4'-bis[4,6-di[4-amilophelyl-N ,N-bis(2-carbamoyethyl)sulfoflYlimifl-I ,3,5-triazin-2- ylamino]-stilbene-2,2'-disulfoflic acid or a pharmaceutically acceptable salt thereof. 4,4'-bis[4 ,6-di[4-aminophenyl-N 1 N-bis(2-carbamoylethyl)sulfoflylimiflo]-l ,3,5-triazin-2-ylamino]- biphenyl-2,2'-disulfonic acid or a pharmaceutically acceptable salt thereof.
13. 16. 4,4'-bis[4,6-di[3-aminophenylsulfoflylamido]-1 ,3,5-triazin-2-ylaminol-stilbefle-2,2'- disulfonic acid or a pharmaceutically acceptable salt thereof.
14. 17. 4,4'-bis[4,6-di[3-aminophenylsulfoflylimido]-1 ,3,5-triazin-2-ylamino]-bipheflyl-2,2'- disulfonic acid or a pharmaceutically acceptable salt thereof.
15. 18. 4,4'-bisl4 1 6-di[3-aminophenyl-N ,N-bis(2-carbamoylethyl)ulfoflylimil- 1 ,3,5-triazin-2- yloxy]-biphenyl-2,2'-disulfonic acid or a pharmaceutically acceptable salt thereof.
16. 19. 5,5'-dimethyl-4,4'-bisl4 ,6-di[3-aminopheflyl-NN-bis(2-carbamoylethyl)ulfoflylimilo]- 1 ,3,5-triazin-2-ylamino]-bipheflyl-2,2'-disulfoflic acid or a pharmaceutically acceptable salt thereof. 4,'bs46d[-mnphnlNNbs2hdoyehlsloyiio ,3,5-triazin-2- ylamino]-biphenyl-2,2'-disulfoflic acid or a pharmaceutically acceptable salt thereof.
17. 21. 9,9-dioxo-2,7-bis[4,6-di[3-aminophelyl-N ,N-bis(2-carbamoylethyl)sulfoflylimilo]-1 S 20 triazin-2-ylamino]-dibenzothiophel-3,6-diulfoflic acid or a pharmaceutically acceptable salt thereof. benzamide]-1 ,3,5-triazin-2-ylaminoI-stilbele-2,2'-disulfoflic acid or a pharmaceutically acceptable salt thereof.
18. 23. 4,'bs46di4aioN-3-mnpey ,N'-bis(2-hydroxyethyl)sulfolylimil- 25 benzensulfonamide]-1 ,3,5-triazin-2-ylamino]-stilbefle-2,2'-disulfoflic acid or a pharmaceutically acceptable salt thereof.
19. 24. 4,4-i-46bs3[i-2craoity)s fmy]pey amn) 1,3, 5-tri azin-2- ylamino]-biphenyl-2,2'-dicarboxylic acid or a pharmaceutically acceptable salt thereof. 4,'bs[46bs[3[bs[-mtyaio)3oorplaiosloy triazi n-2-yl] amino]~-[ 1 '-biphenyl]-2,2'-disuIfoflic acid or a pharmaceutically acceptable salt thereof. *a26. 4,4 i-46bs(-bs(2mtycraolty)sufmy 2-ylamino)-biphenyl-2,2'-dicarboxylic acid or a pharmaceutically acceptable salt thereof.
20. 27. 4,'bs(,-i-3[i-2craoyehl ufmy]peyaio-,3,5-triazin-2- ylamino)-biphenyl-2,2-dicarboxylic acid dimethyl ester or a pharmaceutically acceptable salt thereof.
21. 28. 4,4-i-46bs 3[i-2hdoypoy)slaol peyaio-,3,5-triazin-2- ylamino)biphenyl-2,2'-disulfoflic acid or a pharmaceutically acceptable salt thereof.
22. 29. 4,4-i-46bs -bi-2hdoyrpl-ulaol hnlmnl 1 3,5-triazin-2- yl amino) bi phenyl-2,2'-d icarboxylic acid or a pharmaceutically acceptable salt thereof. 4' ,4-bis-{4 ,6-bis-[3-(bis-carbamoylmethyl-1 -sulfamoyl)-phenylamilo]-1 ,3,5-triazin-2- ylamino}-biphenyl-2,2'-disulfoflic acid or a pharmaceutically acceptable salt thereof .salt thereof. Libc/02390 36
23. 31. 4' ,4-bis-{4,6-bis-[3-(bis-carbamoylmethyl-I -sulfamoyl)-phenylamino]-1 ,3,5-triazin-2- ylamino}-biphenyl-2,2'-dicarboxylic acid or a pharmaceutically acceptable salt thereof.
24. 32. ,2-ethenediyl)bis[5-[[4,6-bis[[3-[[3,5-biS(amiflocarbolyl)-l piperedinyllsulfonyllphenyl]amiloI-l,3,5-triazin-2-yllaminolbenzenesulfonic acid] or a pharmaceutically 5 acceptable salt thereof.
25. 33. 4',4-bis-{4 ,6-bis-[3-(3,5-dicarbamoylpiperidine-1 -sulfonyl)-phenylamino]-1 ,3,5-triazin-2- ylamino}-biphenyl-2,2'-disulfonic acid or a pharmaceutically acceptable salt thereof.
26. 34. 11J, "'-[(2,2'-disulfoll A '-biphenyl]-4,4'diyl)-bis[info-1 ,3,5-triazine-6,2,4-triylbis(imino- 3,1 -phenylenesulfonyl)Itetrakis[3,5-piperidiledicarboxylic acid] or a pharmaceutically acceptable salt thereof. 4,'bs(,-i-3[i-2hdoxehl ufmy]peyaio-,3,5-triazin-2- ylamino)biphenyl-2,2'-dicarboxylic acid or a pharmaceutically acceptable salt thereof.
27. 36. 4,4'-bis[4,6-di[3-aminophel-N Nbis(3-hydroxypropyl)-sufonylimilo]-1 ,3,5-triazin-2- ylamino]-biphenyl-2,2'-disulfoflic acid or a pharmaceutically acceptable -Isalt thereof.
28. 37. A method for treating a viral infection in a mammal, the method comprising administering to the mammal experiencing the viral infection an efficacious amount of a compound of the structure: X N 13-1A B' N X N N NON C. *wherein: A is C 0 C, C0 Cr C'is S0H S3,Oo OH;B sNNIo ;R i ,(lC0oe lyl tagto rnhd whri h abnaos a eotoal usittdwt l r ,O o N sCFo h Libc102390 Z z ,-N Y/Z N N z moiety w U' is S 2 or W' is: Y z -N )m z or z Y is (CH2)n; n is 0 to 6; m is 0 to 2; Z is H, CH 3 CF3, CH2-(halogen), where halogen is CI, Br, F or I, CH20H, -COOH, COO(C1-C6)lower alkyl straight or branched, -CONR 2 R 2 2 R CN or OH and R 2 in each occurrence, is independently H or (C1-C6)lower alkyl; and the pharmaceutically acceptable salts and esters thereof.
29. 38. The method of claim 37 in which the efficacious amount is a dose of from about 10mg/kg to about 500mg/kg.
30. 39. The method of claim 37 or 38 in which the viral infection is a respiratory syncitial virus infection. 10 40. The method of claim 37 or 38 in which the viral infection is a herpes simplex virus infection.
31. 41. The method of claim 37 or 38 in which the viral infection is a human immunodeficiency virus infection.
32. 42. The method of claim 37 or 38 in which the viral infection is a cytomegalovirus infection. 15 43. The method of any one of claims 37 to 42, wherein a compound as claimed in any one of claims 2 to 36 is used.
33. 44. A pharmaceutical composition comprising a compound of the structure: X N B A B' N X N N A NO N Libcd02390 C'C' wherein: A is C. C, C, 0 C C' o- or C' is- SO 3 H, OSO3H, OH, or COOH; B' is NH, NR 1 or O; R 1 is H, (C1-C6)lower alkyl, straight or branched, wherein the carbon atoms may be optionally substituted with Cl, Br, F, OH or CN; X is CI, F or the z Y-Z -N )M 0.0* z z moiety U' is S02, -CO, or W' is: z No or Z Y is (CH2)n; n is 0 to 6; m is 0 to 2; Z is H, CH3, CF3, CH2-(halogen), where halogen is CI, Br, F or I, CH 2 0H, -COOH, COO(C1-C6)lower alkyl straight or branched, -CONR 2 R 2 99 2 R 'OH CN or OH and R 2 in each occurrence, is independently H or (C1-C6)lower alkyl; and the pharmaceutically acceptable salts and esters thereof, and one or more pharmaceutically acceptable carriers. A pharmaceutical composition as claimed in claim 44, containing a compound claimed in any one of claims 2 to 36.
34. 46. A compound as claimed in claim 1, substantially as described herein with reference to any one of the examples.
35. 47. A method as claimed in claim 37, wherein a compound as claimed in claim 46 is administered.
36. 48. A pharmaceutical composition as claimed in claim 44, containing a compound as claimed in claim 46.
37. 49. An antiviral triazine derivative, substantially as hereinbefore described with reference to any one of the examples. Libc/02390 39 A process for the preparation of an antiviral triazine derivative, substantially as hereinbefore described with reference to any one of the examples.
38. 51. Use of a compound of the structure: X N B' B N X A N -rN NON B' B' C. C. S wherein: A is C CC C/ C, C, CC C o or C' is- SO 3 H, OSO 3 H, OH, or COOH; B' is NH, NR 1 or 0; R 1 is H, (Cl-C6)lower alkyl, straight or branched, wherein the carbon atoms may be optionally substituted with Cl, Br, F, OH or CN; X is Cl, F or the z B" N moiety u' w U' is S02, -CO, or W' is: -N z or z Y is (CH 2 n is 0 to 6; m is 0 to 2; Z is H, CH 3 CF 3 CH 2 -(halogen), where halogen is Cl, Br, F or I, CH 2 0H, -COOH, COO(C1-C6)lower alkyl straight or branched, -CONR 2 R2 2 R CN or O H and R2, in each occurrence, is independently H or (C1-C 6 )lower alkyl; or a pharmaceutically acceptable salt or ester thereof for the manufacture of a medicament for treating a viral infection in a mammal. Libc/02390
39. 52. Use of a compound according to any one of claims 2-36, 46 or 49, for the manufacture of a medicament for treating a viral infection in a mammal.
40. 53. Use according to claim 51 or 52, in which the viral infection is a respiratory syncitial virus infection.
41. 54. Use according to claim 51 or 52, in which the viral infection is a herpes simplex virus infection. Use according to claim 51 or 52, in which the viral infection is a human immunodeficiency virus infection.
42. 56. Use according to claim 51 or 52, in which the viral infection is a cytomegalovirus infection.
43. 57. A compound of the structure: X. ,N ,N ,.X C C C, C, wherein: A is C C, C,, C, 0 C C or C' is-SO 3 H, OSO3H, OH, or COOH; B' is NH, NR 1 or O; R 1 is H, (Cl-C6)lower alkyl, straight or branched, wherein the carbon atoms may be optionally substituted with Cl, Br, F, OH or CN; X is Cl, F or the moiety U'W' U' is S02, -CO, or W' is: z Z Y-Z -N -N Y z or Y is (CH 2 n is 0 to 6; m is 0 to 2; Z is H, 0 CH3, CF 3 CH 2 -(halogen), where halogen is CI, Br, F or I, CH20H, -COOH, COO(C 1 -C 6 )lower Libc/02390 2 R alkyl straight or branched, -CONR 2 R 2 CN or OH and R 2 in each occurrence, is independently H or (CI-C 6 )lower alkyl; or a pharmaceutically acceptable salt or ester thereof when used in an efficaceous amount for treating a viral infection in a mammal.
44. 58. A compound according to any one of claims 2-36, 46 or 49, when used in an efficaceous amount for treating a viral infection in a mammal.
45. 59. A compound according to any one of claims 2-36, 46 or 49, when used in an efficaceous amount for treating a respiratory syncitial virus infection in a mammal. A compound according to any one of claims 2-36, 46 or 49, when used in an efficaceous amount for treating a herpes simplex virus infection in a mammal.
46. 61. A compound according to any one of claims 2-36, 46 or 49, when used in an efficaceous amount for treating a human immunodeficiency virus infection in a mammal.
47. 62. A compound according to any one of claims 2-36, 46 or 49, when used in an efficaceous amount for treating a cytomegalovirus infection in a mammal. Dated 16 July 1999 S. 15 AMERICAN CYANAMID COMPANY 0* o SPatent Attorneys for the Applicant/Nominated Person SPRUSON&FERGUSON a *a o Libc/02390 Triazine Containing Compounds Useful as Antiviral Agents Abstract This invention provides triazine ring containing compounds, pharmaceutical compositions containing these compounds and methods utilising these compounds for treating viral; infections, particularly infections by respiratory synctial virus, the compounds having the general structure: N^DN N^DN U, B' B' x U wherein: A is CC1 or C' is- 10 SO 3 H, OSO 3 H, OH, or COOH; B' is NH, NR 1 or O; R 1 is H, (C 1 -C6)lower alkyl, straight or branched, wherein the carbon atoms may be optionally substituted with CI, Br, F, OH or CN; X is CI, F or the 7 Y-z B N Smoiety U' is SO2, -CO, or W' is: -N M &48 z or Y is (CH 2 n is 0 to 6; m is 0 to 2; Z is H, CH 3 CF3, CH2-(halogen), where halogen is CI, Br, F or I, CH 2 0H, -COOH, COO(Ci-C6)lower alkyl straight or branched, -CONR 2 R 2 2 R OH CN or OH and R 2 in each occurrence, is independently H or (C1-C6)lower alkyl; and the pharmaceutically acceptable salts and esters thereof.