MXPA97003286A - Inhibitors of cellular proliferation in muscle l - Google Patents

Abstract

This invention comprises polyanionic benzyl glucosides of triazide benzene amides of formula (I), wherein each of R1, R2, R3, and R4 are, independently, H, SO3M, or (a) and each oligosaccharide group contains 1 to 3 sugar groups, M is lithium, sodium, potassium, or ammonium, n is 1 or 2, X is halogen, lower alkyl having 1 to 6 carbon atoms, or lower alkoxy having 1 to 6 carbon atoms; carbonyl or sulfonyl, or pharmaceutically acceptable salts thereof, as well as their use as inhibitors of cell proliferation in smooth muscle and as therapeutic compositions for treating diseases and conditions characterized by excessive proliferation of muscle.

Description

INHIBITORS OF CELLULAR PROLIFERATION IN SMOOTH MUSCLE FIELD OF THE INVENTION This invention relates to novel polyanionic benzyl glycosides of triazide benzene amides. More particularly, this invention relates to polyanionic benzyl glycosides of triazide benzene amides and their use as inhibitors of cell proliferation in smooth muscle and as therapeutic compositions for the treatment of diseases and conditions that are characterized by excessive proliferation of smooth muscle, such as restenosis.

BACKGROUND OF THE INVENTION The proliferation of smooth muscle cells (CML) is a critical event in the pathogenesis of atherosclerosis and arteriosclerosis of transplants, as well as in the response to damage that arises from all forms of vascular reconstruction such as angioplasty (Raines EW; Ross R. Br. Heart J. 1993, 69 (Supplement), S 30; Clowes, A.; Reidy, MAJ REF: 24539 Vasc. Surg 1991, 13, 885; Isik, F.F .; McDonald, T.O .; Ferguson, M .; Yamanaka, E .; Gordon Am. J. Pa thol. 1992, 141, 1139). However, clinically effective inhibitors of cell proliferation in smooth muscle, for use as anti-stenotic agents, have not been successful to date (Herr an, JPR; Hermans,. RM; Vos, J .; Serruys PW Drugus 1993, 4, 18 and 249. Re-endothelialization of the damaged area, concurrent with cell proliferation in smooth muscle, is a major consideration for inhibiting restenosis (Casscells, Circulation 1992, 86, 722, Reidy, MA, Lidner, V. in Endo Theli al Cel l Dysfuncti ons, Simionescu, N. and Simionescu M., Ed. Plenum Press, NY NY, (1992), 31) Thus, any successful approach to inhibit proliferation in CML should not interfere with the repair of endothelial cells or normal functioning of other cell types (eissberg, PL, Grainger, DJ, Shanahan CM, Metcalfe, JC Cardiovascular Res. 1993, 27, 1191) .The glycosaminoglycans heparin and heparan sulfate are endogenous inhibitors of the proliferation in CML and they are even able to promote endothelial cell growth (Castellot, J.J. Jr; Wright. T.C .; Karnovsky, M. J. Seminars in Thrombosis and Hemostasis 1987, 13, 489; ight, T. N. Arteriesclerosis 1989, 9, 1). However, the full clinical benefits of heparin, heparin fragments, chemically modified heparin, low molecular weight heparins, and other heparin-like anionic polysaccharides may be encompassed by other pharmacological disadvantages (particularly excessive bleeding that arises of anti-regulation effects) coupled with the heterogeneity of the different preparations (Bor an, S. Chemcial and Engineering New, 1993, June 28, 27, Schmid, KM, Preisack, M., Voelker, W., Sujatta M .; Karsch, KR Seminars in Thrombosis and Hemostasis 1993, 19 (Supplement 1), 155; A ann, FW; Neuenschwander, C; Meyer, B. Seminars in Thrombosis and Hemostasis 1993, 19 (Supplement 1), 160; Radhakrishnamurthy, B. Sharma, C; Bhandaru, RR; Berenson, GS; Stanzani, L .; Mastacchi, R. Atherosclerosis, 1986 60, 141; Maffrand, JP, Hervert, MM; Bernart, A.; Defreyn, G .; Delevassee, D; Savi, P., Pinot, JJ, Sampol, J. Seminars in Thrombosis and Hemostasis, 1991, 17 (Supplement 2), 186). Since the anticoagulant effects of many of these agents are independent of the antiproliferative activity in CML, it should be expected that polyanionic agents that are of a more homogeneous composition and of a more defined molecular structure should exhibit a more balanced and desirable profile, with lesser effects. Laterally associated with the anionic polysaccharides mentioned above.

Previous technique WO 92/18546 discloses specific sequences of heparin, which can be obtained in pure form through the synthesis or isolation of heparin fragments, which exhibit an anti-proliferation activity of CML. Beta-Cyclodextrin tetradecasulfate has been described as an inhibitor of cell proliferation in smooth muscle and as an effective inhibitor of restenosis (Weisz, P.B.; Hermann, H. C; Joullie, M. M .; Kumor, K .; Levine, E. M .; Macarak, E. J .; Weiner, D. B. Angiogenesis: Key Principie - Science - Technology - Medicine - Steiner R., Weisz, P.B .; Langer, R. Eds. Birkhauser Verlag, Basel Switzerland, 1992. pg. 107; Hermann, H. C; Okada H.H.; Hozakowska, E .; See you. R. F .; Golden, M. A .; Tomaszewski J. E .; Weisz, P. B .; Barnathan E. S. Arteriosclerosis and Thrombosis 1993, 13, 924; Reilly, C. F .; Fujita, T .; Mcldll, R. C.; Stabilito I. I .; Wai-si E .; Johnson, R. G. Drug Development Research 1993, 29, 137. US Patent No. 5,019,562 describes anionic derivatives of cyclodextrins, for the treatment of pathological conditions associated with cell or tissue growth, undesirable. WO 93/09790 describes antiproliferative polyanionic derivatives of cyclodextrins containing at least 2 anionic residues per carbohydrate residues. EP 312087 A2 and EP 312086 A2 describe antithrombotic and anticoagulant properties of sulfated bisaldonic acid amides. US Patents No. 4,431,636, No. 4,431,637, No. 4,431,638, and No. 4,435,387 describe thioglycoside and oxyaryl glycoside derivatives, polysulphates, as modulators of the complement system. The antiproliferative compounds of the CML of the present invention differ from all those of the prior art in that the CML antiproliferative compounds (a) are benzyne triazide amide benzyl glycosides having no structural similarity to heparin, or sulphated cyclodextrins (b) they contain no more than three contiguous sugar residues (trisaccharides) and (b) they have a defined structure.

DESCRIPTION OF THE INVENTION This invention describes the composition and utility of polyanionic p-benzyl glycosides of triazide benzene amides of formula I wherein each of R 1, R 2, R 3, and R 4 are, independently, H, SO 3 M, or and each oligosaccharide group contains from 1 to 3 sugar groups; M is lithium, sodium, potassium, or ammonium; n is 1 or 2; X is halogen, lower alkyl having 1 to 6 carbon atoms, or lower alkoxy having 1 to 6 carbon atoms; And it is carbonyl or sulfonyl; or a pharmaceutically acceptable salt thereof. It is understood that each of the several occurrences of R in any given molecule of formula I, herein, must be the same or different. Similarly, each of the appearances of R 2, R 3, or R 4 may be the same or different from other occurrences of R 2, R 3, or R 4, respectively. Similarly, each of the different appearances, in the present, of Y in any given molecule and each of the occurrences, in the present, of n in any given molecule may be the same or different from any other occurrence of Y on, respectively. It is also understood that the lower alkyl groups, herein, may be straight or branched chain, and may be for example the methyl, ethyl, propyl, isopropyl, or butyl groups. Similarly, the lower alkoxy groups, herein, may be straight or branched chain, and may be for example methoxy, ethoxy, propoxy, isopropoxy or butoxy groups.

A more preferred aspect of this invention is that of the compounds of formula I: I where: each of R1, R2, R3, and R are, independently, H, SO3M, or and each oligosaccharide group contains from 1 to 2 sugar groups; M is lithium, sodium, potassium, or ammonium; n is 1 or 2; X is halogen, lower alkyl having 1 to 6 carbon atoms, or lower alkoxy having 1 to 6 carbon atoms; And it is carbonyl; or a pharmaceutically acceptable salt thereof. The most preferred compounds of this invention are: Sal tris. { [2-methyl-5- (tetra-O-sulfato-β-gluco-pyranosyloxymethyl) phenyl] amide} dodecasóica of benzene-1, 3-5-tricarboxylic acid, or a pharmaceutically acceptable salt thereof. Come on. { [2-methyl-5- (hepta-O-sulfate-β-cellobiosyloxymethyl) phenyl] amide} heneicosasium of benzene-1, 3-5-tricarboxylic acid or a pharmaceutically acceptable salt thereof. Come on. { [2-Chloro-5- (hepta-O-sulfate-β-D-maltosyloxymethyl) phenyl] -amidajheneicosodate of benzene-1, 3-5-tricarboxylic acid, or a pharmaceutically acceptable salt thereof. Come on. { [2-Chloro-5- (hepta-O-sulfa-to-β-D-cellobiosiloxy-methyl) phenyl] -amidajheneicosodate of benzene-1, 3-5-tricarboxylic acid, or a pharmaceutically acceptable salt thereof. Come on. { [2-Chloro-5- (hepta-O-sulfate-β-D-lactosyloxymethyl) -phenyl] -amidajheneicosodic acid of benzene-1, 3,5-tricarboxylic acid, or a pharmaceutically acceptable salt thereof. Come on. { [3, 5- (tetra-O-sulfate-β-D-glucosiloxy-methyl) phenyl] amide} tetracosodate of benzene-1,3,5-tricarboxylic acid, or a pharmaceutically acceptable salt thereof. Come on. { [3,5-bis- (hepta-O-sulfate-β-D-cello-biosyloxymethyl) -phenyl] amide} tetraaceasodic acid of benzene-1,3,5-tricarboxylic acid, or a pharmaceutically acceptable salt thereof.

PROCESS OF THE INVENTION The compounds of the present invention are prepared according to the general sequence of reactions shown in the following scheme: where R, n, and X are as defined above. Thus, a glycosyl bromide 1 is coupled with a benzyl alcohol 2 in the presence of a catalyst such as a mercuric bromide, mercuric cyanide, silver triflate, or silver perchlorate in an aprotic solvent such as dichloromethane, ether, toluene, or nitromethane, at temperatures ranging from -40 ° C to room temperature, to produce the glycoside 3. Reduction of the nitro group of 3 with a reducing agent such as stannous chloride is carried out in a polar aprotic solvent such as ethyl acetate, at an ambient temperature to reflux, or by catalytic hydrogenation in the presence of a catalyst such as palladium on carbon, provides an aniline compound 4. The coupling of 4 with a triazide benzene chloride or a benzene trisulphonyl chloride, is carried out in the presence of an amine base such as triethylamine or diisopropylethylamine, in an aprotic solvent, such as dichloromethane or tetrahydrate. ofurano. The hydrolysis of any of the acetates groups present in the sugars, with a base such as sodium methoxide in methanol or aqueous sodium hydroxide, in methanol, at a temperature from the ambient to the reflux temperature, and the sulfation of some or all the free hydroxyl groups found in the sugars, with a reagent such as the sulfur trioxide-trimethylamine complex, or the sulfur trioxide-pyridine complex, in a polar aprotic solvent such as dimethylformamide or dimethylsulfoxide, at temperatures ranging from 0 ° C to 100 ° C, it produces the desired compounds I. The invention is also directed to pharmaceutical compositions comprised of an effective amount of one or more of the benzene triazide amide polyanionic benzyl glycosides of this invention, either alone or in combination with excipients (ie, pharmaceutically acceptable materials without pharmacological effect). These compositions are useful for diseases and conditions that are characterized by excessive proliferation of smooth muscle cells, which most frequently arise from vascular reconstructive surgery and transplantation, eg, balloon angioplasty, vascular graft surgery, bypass surgery, coronary arteries, and heart transplant. Other disease states in which vascular proliferation is not desired include hyperfunction, asthma, and congestive heart attack. The compounds of the invention are thus useful for the treatment of these diseases and conditions. The compounds of this invention can be administered systemically, for example by intravenous injection, which typically ranges from 0.1 to 10 mg / kg / h over a range of 5 to 30 days, or by subcutaneous injection at low doses, by administration orally in higher doses than intravenous injection. Localized delivery of the compounds of this invention can be achieved via transmembrane, transdermal, or other topical administration routes, using appropriate, continuous release devices, such as a support matrix, where applicable. The compositions of this invention can be formulated with conventional excipients such as a filler, a disintegrating agent, a binder, a lubricant, a flavoring agent and the like. These are formulated in a conventional manner. It is understood that the compounds of this invention can be administered in any manner and at any concentration that is effective for the particular patient. The manner of supply and the composition and concentration of the dose will be determined on an individual basis, by the physician or other experienced medical professional who is treating the patient.

EFFECTS ON CELLULAR PROLIFERATION A. Cell sources The ability of the compounds of the present invention to inhibit the proliferation of smooth muscle cells and to modulate the growth of endothelial cells was established using isolated aortic cells obtained from commercial sources or, for certain species, prepared at the study site. The cell lines used in this study include aortic, human and porcine smooth muscle cells, and human aortic endothelial cells. Human aortic cell lines were obtained from Clonectics Corporation (San Diego). The porcine aortas were received from a local slaughterhouse. The material froze during transit. The aorta was scrupulously cleaned of the fatty tissue and rinsed in sterile saline solution buffered with phosphates with 2% antibiotic / antifungal (Gibco catalog # 600-5240 AG). The tissue was then digested in a volume of 10 to 15 mL of "Enzyme Mixture" containing type I collagenase, 165 U / mL; Elastase type III, 15 U / mL; BSA, 2 mg / mL; and soybean trypsin inhibitor, 0.375 mg / mL followed by incubation at 37 ° C under 5% CO2, for a time of 10 to 15 minutes. After this treatment, the outer adventitial surface was easily removed by peeling. The aorta was then cut longitudinally and placed in the open position and the endothelial layer was removed by shaving. The xiedia layer of cells was rinsed in enzyme solution, and placed in a new 100 mm dish with 10 mL of enzyme solution. The aorta was crumbled using a pair of fine scissors and digested for 2-3 hours at 37 ° C in 30 mL of freshly prepared enzyme solution. After digestion, the tissue was homogenized using a sterile Pasteur pipette with a burnished tip or an eppendorf pipettor with a sterile pipette tip of 200 to 1000 mL. The suspension was then centrifuged for 10 minutes at 8000 rpm and the pellets were suspended in a volume of 4 to 6 mL of freshly prepared medium and plated on 4 to 6 100 mm flasks with vented lids. The cells were allowed to grow to confluence and divided using 0.25% trypsin. The purity and overall quality of the cells were evaluated using antibody to the actin of the CML.

B. Effects of the compounds on cell proliferation using the incorporation of Thymidine H Cell tests were performed, in an early step (generally a step of 3 to 7) under subconfluence conditions. Cultures were grown in 16-well multi-well culture plates (24 wells) in 199 mediums supplemented with 10% fetal bovine serum and 2% antibiotic / antifungal. In the subconfluence, the cells were placed in a defined serum-free medium (AIM-V, Gibco) for a period of 24 to 48 hours before starting the experimental protocol. Although the compounds were found to be more effective with longer preincubations in general, the experiments were initiated with the addition of the compound thymidine H and serum / growth factor, to synchronized cells devoid of serum, and the results were reported accordingly in this invention. The growth factor and the stimulations with serum were optimized for each type of cell. The compounds were added to each well with a 50-fold dilution (20 μL / well) and the plates were incubated for 24 to 26 hours at 37 ° C in 5% CO2. In this series it was found that all the compounds were soluble in H2O and from here, the test compounds were initially diluted in H2O and were serially diluted in the media. The compounds were routinely assayed at concentrations of 1, 10, and 100 μM. As a control, it is routinely assayed, in all cell preparations, at concentrations of 0.1 to 10 μg / mL, heparin (sodium salt) from porcine intestinal mucosa grade II and Sigma (H-7005). At the end of the experiment the plates were placed on ice, washed three times with ice-cold PBS and incubated in 10% trichloroacetic acid (TCA) for 30 minutes to remove the acid-soluble proteins. The solution was transferred to vials for scintillation containing 0.4N HCl (500 μL / vial, to neutralize the NaOH) and each well was rinsed twice with water (500 μL) for a total volume of 2 mL / vial. The data were obtained, in triplicate, for both the control and the experimental samples. The control data (100%) were obtained from maximally stimulated cells, such as the result of growth factor or serum stimulation. Experimental data were obtained from cells maximally stimulated with growth factor or serum and treated with the compound. The data were expressed as one percent of the control, from which the IC503 values could be determined. The compounds of the present invention are effective inhibitors of smooth muscle cell proliferation, as summarized in Table I. In addition, the compounds of the present invention exhibit an antiproliferative activity of human smooth muscle cells (MHC) in the case where the proliferation is stimulated either by 10% fetal bovine serum(FBS) or by platelet-derived growth factor (FCDP; recombinant human PCDP-AB purchased at Upstate Biotechnology Inc., Lake Placid, NY). For example, the sulfated compound of Example 11 inhibits the proliferation of HSLCs stimulated by the SBF (100 nM IC50) as well as by 5 ng / mL of the FCDP (100 nM IC50).

C. Effect of the growth of endothelial cells versus the proliferation of smooth muscle cells.

The promotion of endothelial cell proliferation, concurrent with the inhibition of smooth muscle cell proliferation, is an important consideration for inhibiting the exaggerated response to damage arising from vascular reconstruction. The compounds of the present invention enhance the growth of human endothelial cells stimulated by 2% FBS at inhibitory doses to human smooth muscle cell proliferation stimulated by 10% FBS, as represented by the sulfated compound of Example 11 as shown in FIG. shows in Figure 1.

D. Cytotoxicity: Visually, all cells were found to tolerate high levels of all compounds fairly well, however to ensure that no toxicity is present, the cytotoxicity of the compounds was examined using a commercial modification of the MTT assay. Briefly, the cells were grown, again, in 24-well plates to a confluence of 70 to 80% and, as before, devoid of serum for 24 to 48 hours, prior to initiation of the experimental protocol. To ensure that the MTT assay verifies the toxicity rather than the proliferation, the cells were incubated with 250 μM of drug in fresh medium, without serum for 24 hours, at 37 ° C, in a humidified incubator with CO2. At the end of the treatment with the compound, MTT indicator dye is added for 4 hours at 37 ° C. The cells are then lysed and aliquots from each well are transferred to a 96-well plate for analysis. Absorbance at a wavelength of 570 nm with a reference wavelength of 630 nm is recorded using a plate reader for ELISA. The results are reported as viable percent without using a drug (100% viable) and presolubilization standards (0% viable). The sulfated compounds of Examples 8-14 do not exhibit toxicity at 250 μM.

E. Anticoagulant activity The anticoagulant activity of the compounds of this invention is evaluated in a partial thromboplastin time (ETTP) assay using normal human plasma collected from 5 donors, using the procedure of Fenichel et. to the. (Clin.Chem. 1964, 10, 69). A timer for the automatic precision coagulation Fibrometro BBL was used using a 0.3 ml probe. Partial thromboplastin activated with ellagic acid was used for these experiments. This reagent is added to human plasma treated with citrates, equilibrated at 37 ° C in a plastic well on the blood clot timer. Calcium is added at 37 ° C, the blood clot timer is run and the time for fibrin formation is recorded (in seconds). The effect of the compounds, added to plasmas, was determined through a concentration of 12.5 - 200 μg / mL. Any plasma that did not clot after 240 seconds was assigned a coagulation time of 240 seconds. A non-fractionated heparin comparator was used, through the concentration range of 1.25 to 10 μg / mL. The coagulation analyzes at all concentrations were carried out in triplicate. An analysis of variance for a randomized block design was used to determine the significance of the observed differences in coagulation times. Potency is reported in relation to heparin where the ratio > 1 indicates a weaker activity in relation to heparin, on a base of μg / mL.

Table 1 na = not analyzed F. Effect in the Restenosis Experimental Model. Sprague-Dawlay rats, males, weighing between 3-0.4 Kg were housed in polycarbonate cages (2 per box) with bed of pieces of corn (ad lib) A dark cycle: light of 12 hr / 12 hr was maintained with light at 7:00 AM). After acclimatization, the rats were randomly assigned to treatment groups (12 per group). The rats were anesthetized with ketamine, acepromazone, and xylazine. The anterior portion of the neck was shaved, an inclusion of 3 cm was made along the midline, and the trachea was exposed by gently dividing the underlying tissues and muscle. A retractor was inserted to hold the muscles and tissues to the side and the common carotid artery was isolated from the surrounding tissue. A 2 cm segment of the right common carotid artery was lifted such that an 11 mm approximator was placed in the vessel at approximately 2 and 7 cm before the bifurcation of the internal and external branches of the carotid artery. The 5 mm segment of the common carotid artery between the approaching forceps was pierced approximately 0.5 mm from the edge of each forceps with a 30 G needle and a gentle stream of isotonic saline was injected through the isolated segment to clean any blood A fixed stream of air, at a flow rate of 30-35 cc / min, was then directed through the artery for 5 minutes to dry the container. The approver initially loosened to remove air from the segment and then removed to restore blood flow. Gentle pressure was applied to the neck until the bleeding stopped. The incision in the neck was sutured, closed and the animal was returned to the individual housing (polycarbonate boxes with bed of pieces of corn). A false operation was performed in a control group of rabbits in which the carotid artery was exsanguinated but not subjected to air drying. Experimental drugs i.v. were administered. from 2 ml ALSA osmotic pumps for 2 weeks starting 2 days before air drying. A 12 mm incision was made along the posterior midline of the rat's neck. To insert the pumps, subcutaneous pockets were formed through the incision along the back of the animal and the pump was implanted in the bag and the attached cannula was pushed subcutaneously to the anterior neck where it was inserted into the jugular vein contralateral to the vessel damaged arterial Two weeks after the surgery, the rats were sacrificed via anesthetized exsanguination. The damaged arterial tissue was fixed in situ by infusion with Formalin Zn2 *, then removed for histological processing and evaluation. The arterial tissue was cut into sections of 5 microns at intervals of 200 to 500 microns. The cross sections were photographed and the arterial and intimal media were digitized and measured. These values were used to determine the intimate relationship / means (I / M); Different groups were compared by ANOVA. The compounds of the present invention have been found to be effective inhibitors of intimal thickening in this experimental model of restenosis. For example, the sulfated compound of Example 11 inhibited intimal thickening by 50% compared to untreated animals at a dose of 125 μg / mL according to the procedure described above. Specific procedures are described in the following examples. These examples are given to illustrate the invention and should not be construed as limiting the invention set forth in the appended claims.

EXAMPLE 1 Step 1 5- (Tetra-O-acetyl-β-glucothyranosyloxymethyl) -2-methyl-1-nitrobenzene A solution of 10.3 g (61.5 mmol) of 4-methyl-3-nitrobenzyl alcohol, 30.3 g (73.7 mmol) of acetobromoglucose, 13.3 g (51.7 mmol) of Hg (CN) is stirred at room temperature for 20 hours. 2, and 18.6 G (52.0 mmol) of HgBr2 in nitromethane (150 mL). The reaction mixture is rapidly cooled with 250 mL of Kbr 2.0 M and then extracted into CH2Cl2- The organic phase is washed with saturated NaC3 solution and brine. The organic phase is dried over MgSO4, concentrated to an oil and semi-purified by flash chromatography using petroleum ether / ethyl acetate (2: 1). Removal of the solvents gives a total of 16.4 g (54% yield) of the title compound as a colorless solid: NMR: H (CDC13 300 MHz) d 7.92 (d, 1 H), 7.43 (dd, 1 H) , 7.33 (d, 1 H), 5.06 - 5.24 (m, 3 H), 4.92 (d, 1 H), 4.66 (d, 1 H), 4.59 (d, 1 H) v, 4.28 (dd, 1 H), 4.18 (dd, 1 H), 3.68 - 3.72 (m, 1 H), 2.60 (s, 3 H), 2.11 (s, 3 H), 2.07 (s, 3 H), 2.03 (s, 3 H), and 2.01 ppm (s, 3 H).

Step 2 5- (Tetra-O-acetyl-β-glucopyranosyl siloxymethyl) -2-methyl phenylamine A solution of 5- (Tetra-O-acetyl-β-glucopyran) siloxymethyl) -2-methyl-nitrobenzene (6.73 g, 13.5 mmol) in EtOAc (150 mL) is stirred at 3 ° C for 3 hours. 21.4 g (94.8 mmol) of SnCl2 dihydrate - The reaction mixture is cooled and then cooled rapidly by adding approximately 450 mL of saturated NaHC 3 solution, diluted with CH 2 Cl 2 and filtered through a flock solka. The organic phase of the filtrate is dried (MgSO 4), and concentrated to give a yellow solid which is washed with a little Et 2 < 3 to provide 3.85 g (63% yield) of the title compound: 1 H NMR (CDC13, 300 MHz) d 7.02 (d, 1 H), 6.64 (s, 1 H), 6.63 (d, 1 H), 5.02 - 5.17 (m, 3 H), 4.79 (d, 1 H), 4.50 - 4.55 (m, 2 H), 4.29 (dd, 1 H), 4.17 (dd, 1 H), 3.64 - 3.68 (m, 1 H), 2.18 (s, 3 H), 2.11 (S, 3 H), 2.02 (s, 3 H), 2.01 (s, 3 H), and 2.00 ppm (s, 3 H).

EXAMPLE 2 Step 1 5- (Hepta-O-acetyl-β-D-cellobiosyloxymethyl) -2-? T? Ethyl- 1-nitrobenzene The title compound is prepared in a yield of 47% by the procedure of step 1 of Example 1, using 4-methyl-3-nitrobenzyl alcohol and acetobromocellobiosei as saturated NaCl solution in the quench step. Purification is achieved by flash chromatography (EtOAc / petroleum ether, 1: 2 to 1: 1 to 2: 1): XH NMR (CDC13, 300 MHz) d 7.91 (s, 1 H), 7.40 (d, 1 H ), 7.32 (d, 1 H), 4.80 - 5.20 (m, 6 H), 4.40 - 4.80 (m, 4 H), 4.36 (dd, 1 H), 4.02 - 4.13 (m, 2 H), 3.81 ( t, 1 H), 3.60 - 3.68 (m, .2 H), 2.60 (s, 3 H), 2.14 (s, 3 H), 2.08 (s, 3 H), 2.06 (s, 3 H), 2.03 (s, 3 H), 2.02 (3, H), 2.01 (s, 3 H), and 1.98 ppm (s, 3 H).

Step 2 5- (Hepta-O-acetyl-β-D-cellobiosyloxymethyl) -2- methylpheni sheet The title compound m.p. 180-182 ° C, is prepared with a 62% yield from 5- (Hepta-O-acetyl-β-D-cellobiosyloxymethyl) -2-methyl-1-nitrobenzene, using the procedure of step 2 of Example 1. Purification is achieved by flash chromatography (EtOAc / petroleum ether, 1: 1 to 2: 1): XH NMR (DMSO-d6, 400 MHz) d 6.86 (d, 1 H), 6.48 (dd, 1 H), 6.36 (dd, 1 H), 5.24 (t, 1 H), 5.10 (m, 1 H), 4.80 - 4.90 (m, 3 H), 4.61 - 4.72 (m, 2 H) , 4.56 (d, 1 H), 4.30 - 4.31 (m, 2 H), 4.22 (dd, 1 H), 4.08 (dd, 1 H), 3.99 - 4.03 (m, 1 H), 3.94 (dd, 1 H), 3.73 -3.81 (m, 2 H), 2.10 (s, 3 H), 2.10 (s, 3 H) 2.00 (s, 3 H), 1.98 (s, 3 H), 1.96 (s, 3 H), 1.95 (s, 3 H), 1.94 (s, 3 H), and 1.91 ppm (s, 1 H); mass spectrum (+ FAB) m / z 778. Analysis calculated for C34H46NO18: C, 53.97; H, 6.13; N, 1.85. Found: C, 53.67; H, 5.92; N, 1.62.

EXAMPLE 3 Step 1 5- (Hepta-O-acetyl-β-D-cellobiosyloxymethyl) -2-chloro-1-nitrobenzene The title compound is prepared in 45% yield by the procedure described in step 1 of Example 1, using 4-chloro-3-nitrobenzyl alcohol and acetobromocellobiose, as well as saturated NaCl solution in the step of quenching: NMR? H (CDC13, 300 MHz) d 7.82 (d, 1 H), 7.53 (d, 1 H ), 7.42 (dd, 1 H), 4.8 - 5.2 (m, 5 H), 4.73 (d, 1 H), 4.51 -4.66 (m, 4 H), 4.3 - 4.4 (m, 1 H), 4.03 - 4.11 (m, 2 H), 3.81 (t, 1 H), 3.60 - 3.68 (m, 2 H), 2.13 (s, 3 H), 2.09 (s, 3 H), 2.06 (s, 3 H), 2.03 (s, 3 H), 2.01 (s, 3 H), 2.01 (s, 3 H), 1.99 (s, 3 H), and 1.57 ppm (s, 3 H).

Step 2 5- (Hepta-O-acetyl-β-D-cellobiosyloxymethyl) -2-chloro-phenylamine The title compound is prepared from 5- (hepta-O-acetyl-β-D-cellobiosyloxymethyl) -2-chloro-l-nitrobenzene, in 61% yield, as a solid, by trituration with ether using the procedure of step 2 of Example 1: 1 H NMR (CDCl 3, 300 MHz) d 7.20 (d, 1 H), 6.75 (s, 1 H), 6.60 (d, 1 H), 4.89 - 5.19 (m, 5 H) , 4.73 (d, 1 H), 4.47 - 4.60 (, 4 H), 4.36 (dd, 1 H), 4.02 - 4.13 (m, 2 H), 3.80 (5, 1 H), 3.55 - 3.67 (m, 2 H), 2.14 (s, 3 H), 2.08 (s, 3 H), Z.03 (s, 3 H), 2.02 (s, 6 H), 2.01 (s, 3 H), and 2.00 ppm ( s, 3 H).

EXAMPLE 4 Step 1 5- (Hepta-Q-acetyl-β-D-maltosiloxy ethyl) -2-chloro-l-nitrobenzene The title compound is prepared in 50% yield, by the procedure described in step 1 of Example 1, using 4-chloro-3-nitrobenzyl alcohol and acetobromomaltose, as well as saturated NaCl solution, in the rapid cooling step . Purification is achieved by flash chromatography (CH2CI2: EtOAc (5: 1) to proportional title compound: partial 1H NMR (CD13, 300 MHz) d 7.83 (d, 1 H), 7.53 (d, 1 H), 7.42 (dd, 1 H), 5.42 (d, 1 H), 5.36 (t, 1 H), 5.06 (t, 1 H), 2.15 (s, 3 H), 2.11 (s, 3 H), 2.04 (s) , 3 H), 2.03 (s, 3 H), 2.02 (s, 3 H), and 2.01 ppm (s, 3 H).

Step 2 5- (Hepta-O-aceti l-ß-D-maltos i loximeti l) -2-chloro-l-phenylamine The title compound is prepared from 5- (Hepta-O-acetyl-β-D-maltosiloxymethyl) -2-chloro-1-nitrobenzene in 96% yield, by the procedure of step 2 of Example 1. crude product is obtained as a solid and used without further purification: partial H NMR (CDI3, 300 MHz) d 7.20 (d, 1 H), 6.73 (d, 1 H), 6.60 (dd, 1 H), 5.41 ( d, 1 H), 5.30 - 5.39 (m, 2 H), 5.23 (t, 1 H), 5.05 (t, 1 H), 4.83 - 4.89 (m, 2 H), 4.74 (d, 1 H), 2.16 (s, 3 H), 2.11 (s, 3 H), 2,032 (s, 3 H) 2,027 (s, 3 H), and 200 ppm (s, 6 H).

EXAMPLE 5 Step 1 5- (Hepta-Q-acetyl-β-D-lactosyloxymethyl) -2-chloro-l-nitrobenzene The title compound is prepared in 51% yield by the procedure described in step 1 of Example 1, using 4-chloro-3-nitrobenzyl alcohol and acetobromolactose, as well as saturated NaCl solution, in the quench step. Purification is achieved by initial flash chromatography (CH2CI2: EtOAc (4: 1) and then with a flash chromatography (CH2Cl2 EtOAc (9: 1)) to proportionally give the title compound as a solid: Partial XH NMR (CD13) : 300 MHz) d 7.82 (d, 1 H), 7.53 (d, 1 H), 7.42 (dd, 1 H), 5.35 (d, 1 H), 5.21 (s, 1 H), 4.87 (d, 1 H), 4.66 (d, 1 H), 2.15 (s, 3 H), 2.13 (s, 3 H), 2.06 (s, 6 H), 2.05 (s, 6 H), and 1.97 ppm (s) , 3 H).

Step 2 5- (Hepta-O-acetyl-β-D-lactosyloxymethyl) -2-chloro-phenylamine The title compound is prepared with 5- (Hepta-O-acetyl-β-D-lactosyloxymethyl) -2-chloro-1-nitrobenzene in 94% yield, by procedure 1 of step 2 of Example 1. The product crude is obtained as a solid and used without further purification: NMR 1H partial (CDC13; 300 MHz) d 7.19 (d, 1 H), 6.72 (d, 1 H), 6.59 (dd, 1 H), 5.35 (d, 1 H), 5.08 - 5.20 (m, 2 H), 4.73 (d, 1 H), 2.15 (s, 3 H), 2.14 (s, 3 H), 2.06 (s, 3 H), 2.05 (s, 3 H), 2.02 (s, 3 H), and 1.97 ppm (s, 3 H).

EXAMPLE 6 Step 1 3, 5-Bis- (hepta-O-acetyl-β-D-cellobiosyloxymethyl) -1- nitrobenzene The title compound is prepared in 42% yield, by the procedure described in step 1 of Example 1, using 1 equivalent of 5-nitro-m-xylene-a, a'-diol, 2 equivalents of acetobromocelobiose, and 2 equivalents of all other reagents, as well as saturated NaCl solution, in the rapid cooling step. Purification is achieved initially by flash chromatography (EtOAc / CH2CI2 (3: 1)) and then by trituration in ether: 1E-NMR (CDCl3, 300 MHz) d 8.10 (s, 2 H), 7.48 (s, 1 H) , 4.88 - 5.30 (m, 12 H), 4.68 (d, 2 H), 4.52 -4.59 (m, 6 H), 4.38 (dd, 2 H), 4.0 - 4.13 (m, 4 H), 3.82 (t , 2 H), 3.58 - 3.68 (m, 4 H), 2.13 (s, 6 H), 2.08 (s, 6 H), 2.07 (s, 6 H), 2.03 (s, 12 H), 2.01 (s) , 6 H), and 1.98 ppm (s, 6 H).

Step 2 3, 5-Bis- (hepta-O-acetyl-β-D-cellobiosyloxymethyl) phenylamine The title compound is prepared in 54% yield, by the procedure of step 2 of Example 1, using 3,5-bis (β-D-cellobiosyloxymethyl) -1-nitrobenzene. Purification is accomplished using EtOAc / petroleum ether (1: 1): H NMR (CDCl 3, 300 MHz) d 6.73 (broad 1 H), 6.67 (s. broad 1 H), 5.03 -5.30 (m, 6 H), 4.97 (d, 2 H), 4.91 (d, 2 H), 4.75 (d, 2 H), 4.49 - 4.61 (6 H), 4.37 (dd, 2 H), 4.02 - 4.13 (m, 6 H), 3.81 (t, 2 H), 3.57 - 3.69 (m, 4 H) 2.15 (s, 6 H), 2.08 (s, 6 H), 2.03 (s, 6 H), 2.01 (s, 18 H), and 1.98 ppm (s, 6 H).

EXAMPLE 7 Step 1 3, 5-Bis- (tetra-O-acetyl-β-D-glucosiloxylmethyl) -1- nitrobenzene The title compound is prepared by the procedure described in step 1 of Example 1, using 1 equivalent of 5-nitro-m-xylene-a, a'-diol, two equivalents of acetobromoglucose, and two equivalents of all other reagents , as well as saturated solution of NaCl, in the step of rapid cooling. Purification is achieved by flash chromatography (EtOAc / CH2Cl2 (1: 4 to 1: 2) to give a 29% yield of the almost pure title compound, which is used without further purification.

Step 2 3, 5-Bis- (tetra-O-acetyl-β-D-glucosyloxymethyl) phenylamine The title compound is prepared by the procedure of step 2 of Example 1 using 5.29 g of 3,5-bis (tetra-O-acetyl-β-D-glucosyloxymethyl) -1-nitrobenzene. Purification is achieved by flash chromatography (CH2Cl2 / EtOAc (3: 2)) to give 2.00 g (39% yield) of the title compound as a yellow solid: partial H NMR (CDC13, 300 MHz) d 6.56 ( s, 3 H), 5.0 - 5.2 (m, 6 H), 4.8 (d, 2 H), 4.4 - 4.6 (m, 4 H), 3.7 ppm (very large signal, 2 H).

EXAMPLE 8 Step 1 Benzene-1, 3, 5-tricarboxylic acid Tris. { [2-methyl-5- (β-glucopyranosyloxymethyl) phenyl] amide} To a solution of 5- (tetra-O-acetyl-β-glucopyranosyloxymethyl) -2-methylphenylamine (1.51 g, 3.22 mmol) and 3.22 mmol of triethylamine in THF (35 ml) are added 285 mg (1.01 mmol) of sodium chloride. Benzene-1,3,5-tricarboxylic acid. The reaction mixture is stirred for 4 hours, quenched with MeOH, diluted with CH2Cl2, and washed with H2O. The organic phase is dried (MgSO 4) and concentrated to an oil which is purified by flash chromatography (CHCl: EtOAc (1: 1) to give the crude product (1.17 g, 70% yield). 983 mg (0.631 mmol) of the crude material, in MeOH (25 mL), is added 1N NaOH (9.47 mL, 9.47 mmol) After stirring at 50 ° C for 3 hours, the reaction mixture is rapidly cooled with HCl 1 N (7.58 mL, 7.58 mmol) and the product is isolated by filtration Drying in vacuo gives 700 mg (100% yield) of the crude title compound The solid is suspended in H2O to remove the additional salts and provide the pure title compound, mp> 200 ° C: XH NMR (DMSO-d6, 400 MHz) d 10.3 (s, 3 H), 8.73 (s, 3 H), 7.37 (s, 3 H), 7.28 (d , 3 H), 7.23 (d, 3 H), 5.10 (broad s, 3 H), 4.84 (d, 3 H), 4.57 (d, 3 H), 4.25 (d, 3 H), 3.69 (dd) , 3 H), 3.44 - 3.49 (m, 3 H), 3.0 - 3.2 (m, 6 H), and 2.28 ppm (s, 9 H); 13 C NMR (DMSO-d6; 100 MHz) d 164.6, 136.1, 135.3, 133.0, 130.3, 129.8, 126.0, 125.7, 102.1, 77.0, 76.8, 73.6, 70.2, 69.2, 61.2, and 17.8 ppm; mass spectrum (-FAB) m / e 1052.3, 890.3; IR (KBr) 1650 cm-1. Analysis calculated for C51H63 3O21. 4 H2O: C, 54.40; H, 6.35; N, 3.73. Found: C, 54.44; H, 6.15; N, 3.68.

Step 2 Dodecasodic salt of benzene-1,3,3-Tricarboxylic acid Tris. { [2-methyl-5- (tetra-Q-sulfate-β-glucopyranosyloxymethyl) phenyl] amide} A solution of benzene-1, 3, 5-tricarboxylic acid tris- is stirred at 70 ° C for 4 days. { [2-methyl-5- (β-glucopyranosyloxymethyl) phenyl] -amide} (761 mg, 0.748 mmol) and sulfur trioxide (6.24 g, 44.9 mmol) in DMF (30 mL). The reaction mixture is cooled rapidly to room temperature with H2O and concentrated. Purification is achieved by chromatography with Shepadex G-10 (elution with H2O) followed by cation ion exchange using a Dowex 50 x 8 column of strongly acid resin (of the Na form) to provide 825 mg (48% yield) of the title compound as a colorless solid, mp > 200 ° C: H NMR (D20, 400 MHz) d 8.73 (s, 3 H), 7.46 - 7.51 (broad m, 9 H), 4.97 - 5.00 (m, 6 H), 4.87 (d, 3 H), 4.76 (t, 3 H), 4.47 - 4.53 (m, 6 H), 4.22 - 4.27 (m, 3 H), 4.15 - 4.19 (m, 3 H), and 2.37 ppm (s, 9 H); 13 C NMR (D 20; 100 MHz) d 168.3, 135.3, 135.2, 135.0, 134.3, 131.1 ,. 130.0, 128.1, 127.0, 99.1, 76.2, 75.9, 73.4, 72.6, 70.7, 67.8, and 16.8 ppm; mass spectrum (electroroated) (m-zNa) / z 736.5 (m-3 Na) ", 546.2 (m - 4 Na) 4-, and 433.4 (m - 5 Na) 5-. Analysis calculated for C5? H5iN3Nai2? 5iSi2. 12 H20: C, 24.55; H, 3.01; N, 1.68; S, 15.40 Found: C, 24.29; H, 2.78; N, 2.69; S, 15.84.

EXAMPLE 9 Step 1 Benzene-1, 3, 5-tricarboxylic acid Tris. { [2-methyl-5- (ß-cellobiosyloxymethyl) phenyl] amide} To a solution of 5- (tetra-O-acetyl-β-cellobiosyloxymethyl) -2-methylphenylamine (1.49 g, 1.97 mmol) and 217 μL (1.97 mmol) of triethylamine in THF (35 mL) are added 174 mg ( 0.66 mmol) of benzene-1,3,5-tricarboxylic acid chloride. The reaction mixture is stirred for 4 hours, quenched with MeOH, diluted with CH2Cl2, and washed with H2O. The organic phase is dried (MgSO.sub.4) and concentrated to an oil which is purified by trituration with ether to give 873 mg. 55%) of a whitish solid. To a solution of the crude material in MeOH (25 mL) is added 1 N NaOH (9.1 mL, 9.1 mmol). After stirring at 50 ° C, for 3 hours, the reaction mixture is rapidly cooled with 1 N HCl (7.64 mL, 7.64 mmol) and the product isolated by filtration. Drying in vacuo gives 408 mg (80% yield) of the title compound. p.f. > 200 ° C: Partial 1R NMR (DMS0-d6, 400 MHz) d 10.33 (s, 3 H), 8.75 (s, 3 H), 7.36 (s, 3 H), 7.27 (d, 3 H), 4. 23 (d, 3 H), 5.2 - 5.3 (wide signal, 6 H), 5.00 (broad signal, 6 H), 4.84 (d, 3 H), 4.70 (s, 3 H), 4.56 - 4.64 (m, 9 H), 4.33 (d, 3 H), 4.26 (d, 3 H), 3.77 (dd, 3 H), 3.64 - 3.70 (, 6 H), and 2.27 ppm (s, 9 H): 13 C NMR (DMSO-d6; 100 MHz) d 164.5, 136.0, 135.9, 135.1, 133. 0, 130.2, 129.8, 125.9, 125.6, 103.2, 101.8, 80.5, 76. 8, 76.4, 75.0, 74.9, 73.3, 73.2, 70.0, 69.4, 61.0, 60.4 and 17.8 ppm; mass spectrum ((-) - FAB) 1539.2 ().

Step 2 Heneicosasodic salt of benzene-1,3,3-Tricarboxylic acid Tris. { [2-methyl-5- (hepta-O-sulfate-β-cellobiosyloxymethyl) phenyl] amide} The solution is stirred at 70 ° C for 4 days. 279 mg (149 mmol) of benzene-1,3,5-tricarboxylic acid tris. { [2-methyl-5- (ß-cellobiosyloxymethyl) phenyl] amide} and trimethylamine sulfur trioxide complex (3.19 g) in DMF (30 mL). The reaction mixture is cooled rapidly, at room temperature, with H 2 O and concentrated in vacuo. The residue is purified by Shepadex G-10 chromatography (elution with H 2 O). Cation exchange is effected using a Dowex 50 x 8 strongly acidic resin column (of the Na form) to provide 357 mg of the title compound as a tan solid, m.p. > 200 ° C: partial H-NMR (D20, 400 MHz) d 8.70 (s, 3 H), 7.44-7.52 (m, 9 H), 4.99 (d, 3 H), 4.97 (d, 3 H), 4.86 ( d, 3 H), 4.83 (d, 3 H) 4.68 (d, 3 H), 4.64 (d, 3 H), 4.58 (dd, 3 H), 4.01 -4.04 (m, 6 H), and 2.34 ppm (s, 9 H); 13C NMR (D20; 100 MHz) d 168.17, 135.4, 135.0, 134.9, 134.2, 131.0, 129.9, 127.9, 126.7, 99.9, 99.2, 77.6, 77.34, 77.31, 77.02, 74.3, 73.6 73.4, 73.0, 70.6, 67.6. 66.5, and 16.7 ppm; mass spectrum (electrorored) (m - z Na) z 897.8 (m - 4 Na) 4-, 713.2 (m - 5 Na) 5-, and 590.8 (m - 6 Na) ~. Analysis calculated for: C69H 2N3Na2iOg9S2i • 18 H2O: C, 20.67; H, 2.70; N, 1.05; S, 16.78. Found: C, 20.35; H, 2.78 N, 1.00; S, 14.19.

EXAMPLE 10 Step 1 Benzene-1,3,5-tricarboxylic acid Tris. { [5- (hepta- O-acetyl-β-D-maltosyloxymethyl) -2-chlorophenyl] amide} To a solution of 962 mg (1.24 mmol) of 5- (hepta-O-acetyl-β-maltosyloxymethyl) -2-chlorophenylamine in THF (20 mL) containing 173 μL (1.24 mmol) of triethylamine is added 110 mg ( 0.413 mmol) of benzene-1 acid chloride, 3, 5-tricarboxylic. The reaction mixture is stirred for 90 minutes, quenched with MeOH, diluted with CH2Cl2 and washed with H2O. The organic phase is dried (MgSO 4) and concentrated to yield a light yellow solid. Purification by flash chromatography (CH2CI2: EtOAc (2: 1 to 1: 1) gives 607 mg (59% yield) of the title compound as a colorless solid, mp 132 ° C (dec): Partial H NMR (CDCl 3 , 400 MHz) d 8.66 (s, 3 H), 8.65 (s, 3 H), 8.48 (d, 3 H), 7.42 (d, 3 H), 7.08 (dd, 3 H), 5.40 (d, 3 H), 5.33 (t, 3 H), 5.23 (t, 3 H), 5.03 (t, 3 ñ, 4.82 - 4.93 (m, 9 H), 4.66 (d, 3 H), 4.62 (d, 3 H) ), 4.53 (dd, 3 H), 3.67 - 3.70 (m, 3 H), 2.14 (s, 3 H), 2.10 (s, 3 H), 2.04 (s, 3 H), 2.01 (s, 3 H) ), 1.99 (s, 3 H), 1 995 (s, 3 H), 1, 986 (s, 3 H), and 1. 97 ppm (s, 3 H): 13 C NMR (CDC13; 100 MHz) d 170.5; 170.2, 169.9, 169.7, 169.4, 162.9, 137.0, 136.1, 134.1, 129.2, 128.9, 124.2, 123.1, 121.1, 99.1, 95.5, 75.4, 72.6, 72.2, 72.0, 70.1, 70.0, 69.3, 68.5, 68.0, 62.8, 61.5, 20.9, 20.7, 20.6, and 20.5 ppm; mass spectrum (positive electroroded (Ca2 +)) 1262 (m + Ca2 +) / 2. Analysis calculated for C? O8Hi26 l3N3? 5. 1 H2O: C, 51.83; H, 5.16; N, 1.68. Found: C, 51.64; H, 5.11; N, 1.53.

Step 2 Benzene-1, 3, 5-tricarboxylic acid Tris. { [2-chloro-5- (β-D-maltosyloxymethyl) -phenyl] amide} To a solution of 983 mg (0.194 mmol) of benzene-1,3,5-tricarboxylic acid tris. { [5- (hepta-O-acetyl-β-D-matasyloxymethyl) -2-chlorophenyl] amide} in MeOH (15 mL) is added 1N MeOH (4.67 mL, 4.67 mmol). After stirring at 50 ° C for 4 hours, the reaction mixture is quenched with 1 N HCl (4.08 mL, 4.08 mmol) and the product isolated by filtration. Drying in vacuo gives 309 mg (99% yield) of the title compound, m.p. > 200 ° C: 1U partial NMR (DMSO-d6, 400 MHz) d 10.56 (s, 3 H), 8.78 (s, 3 H), 7.60 (d, 3 H), 7.56 (d, 3 H), 7.37 ( dd, 3 H), 5.02 (d, 3 H), 4.89 (d, 3 H), and 4.63 ppm (d, 3 H); 13 C NMR (DMSO-d6; 100 MHz) d 164.57, 137.83, 134.73, 134.61, 130.13, 129.36, 128.67, 127.53, 126.85, 102.12, 100.75, 79.54, 76.37, 75.28, 73.45, 73.24, 73.02, 72.42, 69.87, 68.72 , 60.76, and 60.64 ppm; mass spectrum ((-) - FAB) m / z 1598.4 (M - H), 1436.3 and 1274.3. Analysis calculated for C66H84CI3N3O36. 9 H2O: C, 44.94; H, 5.83; N, 2.38. Found: C, 44.48; H, 5.46; N, 2.78.

Step 3 Heneicosasodic salt of benzene-1,3,5-tricarboxylic acid Tris. { [2-chloro-5- (hepta-O-sulfato-β-D-maltosiloxymethyl) phenyl] amide} A solution of benzene-1, 3, 5-tricarboxylic acid tris- is stirred at 70 ° C for 2.5 days. { [2-chloro-5- (β-D-maltosyloxymethyl) phenyl] amide} (208 mg, (130 mmol)) and sulfur trioxide trimethylamine complex (1.90 g) in DMF (20 mL). The reaction mixture is cooled rapidly, at room temperature, with H20 and concentrated in vacuo. The residue is purified by chromatography on Sephadex G-10 (elution with H O). Cation exchange is effected using a Dowex 50 x 8 hard acid resin column (of the Na form) to provide 400 mg (82% yield) of the title compound as a tan solid, m.p. > 178 ° C (with decomposition): 1H partial NMR (D20, 400 MHz) d 8.74 (s, 3 H), 7.67-7.70 (m, 6 H), 7.57 (d, 3 H), 5.62 (d, 3 H) ), 5.09 (d, 3 H), 5.04 (d, 3 H), 4.66 (t, 3 H), 4.61 (dd, 3 H), - and 4.52 ppm (t, 3 H); 13C NMR (D20; 100 MHz) d 168.1, 136.9, 134.7, 132.9, 130.3, 130.1, 129.7, 128.6, 127.6, 98.8, 94.1, 77.4, 76.0, 74.8, 73.4, 73.2, 72.3, 71.8, 69.9, 69.7, 67.6 , and 66.1 ppm; mass spectrum (electrorored) (m - z) Na) / z 12225.0 (m-3 Na) 3-, 912.8 (m-4 Na) 4-, 725.6 (m-5 Na) ~, and 601 (m-6 Na) ~. Analysis calculated for C66H63Cl3 3Na2i? 99S2i-4 Na2SO4. 21 H20: C, 16.90; H, 2.25; N, 0.90, S, 17.09. Found: C, 16.77; H, 2.29; N, 0.97; S, 16.90.

EXAMPLE 11 Step 1 Benzene-1,3,5-tricarboxylic acid Tris. { [5- (hepta- O-acetyl-β-D-cellobosyloxymethyl) -2-chlorophenyl] amide} To a solution of 1.43 g (1.85 mmol) of 5- (hepta- O-acetyl-β-cellobiosyloxymethyl) -2-chlorophenylamine in THF (25 mL) containing 254 μL (1.85 mmol) of triethylamine, 163 mg (0.615 mmol) of benzene-1,3,5-tricarboxylic acid chloride are added. The reaction mixture is stirred for 90 minutes, quenched with MeOH, diluted with CH2Cl2, and washed with H2O. The organic phase is dried (MgSO4) and concentrated to an off-white solid. Purification by flash chromatography (CH2Cl2: EtOAc (1: 1)) gives 937 mg (62% yield) of the product. An analytical sample, obtained as a white sample (mp> 200 ° C), is prepared from a separate assay: 13 C NMR (CDC13, 100 MHz) d 170.5, 170.4, 170.2, 169.8, 169.6, 169.3, 169.0, 163.0, 137.0, 136.1, 134.0, 129.2, 128.9, 124.7, 123.2, 121.2, 100.75, 99.4, 76.37, 72.90, 72.8, 72.47, 71.93, 71.59, 71.46, 70.1, 67.8, 61.8, 61.5, 20.87, 20.66, 20.60, and 20.50 ppm; mass spectrum (positive electroroded (Ca2 +)) (m + Ca) 2 + / 2 1262. Analysis calculated for C108H126CI3N3O57 • 2H20: C, 51.46; H, 5.20; N, 1.67. Found: C, 51.44; H, 4.88; N, 1.71.

Step 2 Benzene-1, 3, 5-tricarboxylic acid Tris. { [-2-chloro-5- (ß-D-cellobiosyloxymethyl) phenyl] amide} A solution of 917 mg (0.375 mmol) of benzene-1,3,5-tricarboxylic acid tris. { [5- (hepta-O-acetyl-β-D-cellobiosyloxymethyl) -2-chlorophenyl] amide} in MeOH (25 L), treated with 9.0 mL (9.0 mmol) of 1 N NaOH. After stirring for 3 hours at 50 ° C, the reaction mixture is cooled rapidly, at room temperature, with 1 N HCl (7.9). mL, 7.9 mmol) and stirred for 10 minutes. The solid is combined and dried in vacuo to provide 579 mg (96% yield) of the title compound as a colorless solid, m.p. > 200 ° C: 1H partial NMR (DMSO-d6 / 400 MHz) d 10.57, (s, 3 H), 8.78 (s, 3 H), 7.58 (s, 3 H), 7.55 (d, 3 H), 7.36 (s, 3 H), 4.87 (d, 3 H), 4.34 (d, 3 H), 4.25 (d, 3 H), 3.77 (d, 3 H), and 3.68 ppm (d, 3 H); 13 C NMR (DMSO-d6; 100 MHz) d 164.6, 137.9, 134.7, 134.6, 130.2, 129.4, 128.7, 127.5, 126.9, 103.2, 102.0, 80.5, 76.8, 76.5, 75.0, 73.3, 73.2, 70.0, 68.8, 61.0 , and 60.4 ppm; IR (KBr) 1655 cm "1; mass spectrum (+ FAB) m / z 1622.2 (M + Na) Analysis calculated for C66H84 I3N3O36.5H 0: C, 46.86; H, 5.60; N, 2.48. C, 46.84; H, 5.46; N, 2.37.

Step 3 Heneicosasodic salt of benzene-1,3,5-tricarboxylic acid Tris [2-chloro-5- (hepta-O-sulfo-β-D- c_ Lobiosyloxymethyl) phenyl] amide) A solution of 403 mg (0.252 mmol) of tris benzene-1,3,5-tricarboxylic acid is stirred at 70 ° C for 5 days. { [2-chloro-5- (β-D-cellobiosiloxy ethyl) phenyl] amide} and trimethyl amine sulfur trioxide complex (3.89 g, 28 mmol) in DMF (25 mL). The reaction mixture is cooled rapidly, at room temperature, with H2O and concentrated in vacuo. The residue is purified by chromatography with Sephadex G-10 (elution with H2O). The cation exchange is effected using a Dowex 50 x 8 strongly acidic resin column (of the Na form) to provide 949 mg of the title compound as a tan solid, mp. 178 ° C (with decomposition): Partial NMR * H (D20, 400 MHz) d 8.73 (s, 3 H), 7.70 (s, 3 H), 7.67 (d, 3 H), 7.57 (d, 3 H) , 4.99 - 5.03 (m, 6 H), 4.68 (t, 3 H), 4.64 (t, 3 H), 4.60 (dd, 3 H), 4.24 (d, 3 H), 4.20 (t, 3 H) , and 4.02-4.05 ppm (m, 3 H); 13C NMR (D20; 100 MHz) d 168.1, 139.0, 134.7, 132.8, 130.3, 130.0, 129.6, 128.5, 127.4, 99.9, 99.4, 77.5, 77.4, 77.3, 77.0, 74.3, 73.7, 73.3, 73.1, 70.1, 67.6 , and 66.6 ppm. Analysis calculated for C66H63Cl3 3Na2i? 99.33 H20: C, 18.28; H, 2.93; N, 0.97; S, 15.53. Found: C, 18.20; H, 2.67; N, 0.76; S, 15.51. Capillary electroporosis shows a purity of more than 98%.

EXAMPLE 12 Step 1 Benzene-1, 3, 5-tricarboxylic acid Tris. { [2-chloro-5- (β-D-lactosyloxymethyl) phenylamide} To a solution of 993 mg (1.28 mmol) of 5- (hepta-O-acetyl-β-lactosyloxymethyl) -2-chloro-phenylamine in THF (20 mL) containing 178 μL (1.28 mmol) of triethylamine is added. mg (0.427 mmol) of benzene-1,3,5-tricarboxylic acid chloride. The reaction mixture is stirred for 90 minutes, quenched with MeOH, diluted with CH2CI2 / and washed with H2O. The organic phase is dried (MgSO 4) and concentrated to an off-white solid. Purification by flash chromatography (CH2Cl2: EtOAc (2: 1 to 1: 1)) gives 431 mg (41% yield) of the product. A solution of 431 mg (0.174 mmol) of the compound in MeOH (15 mL) is treated with 4.2 mL (4.2 mmol) of 1 N NaOH. After stirring for 4 hours at 50 ° C, the reaction mixture is rapidly cooled to Room temperature with IN HCl (3.6 mL, 3.6 mmol). The solid is combined and dried in vacuo to provide 193 mg (yield 69%) of the title compound as a colorless solid, m.p. > 200 ° C: partial XH NMR (DMSO-d6, 400 MHz) d 10.58 (s, 3 H), 8.79 (s, 3 H), 7.59 (d, 3 H), 7.56 (d, 3 H), 7.36 (dd, 3 H), 4.88 (d, 3 H), 4.63 (d, 3 H), 4.35 (d, 3 H), 4.20 (d, 3 H), 3.79 (d, 3 H), and 3.11 (s, 3 H), 13 C NMR (DMSO-d6, 100 MHz) d 164.6, 137.8, 134.7, 134.6, 130.2, 129.4, 128.7, 127.5, 126.8, 103.8, 102.0, 80.7, 75.5, 75.0, 74.9, 73.22, 73.19, 70.5, 68.8, 68.1, 60.5, and 60.3 ppm; mass spectrum ((-) - FAB) m / z 1598.8 and 1600.8 (both N-H). Analysis calculated for C66H84CI3N3O36. 9 H20: C, 44.94; H, 5.83; N, 2.38. Found: C, 44.24; H, 5.38; N, 2.29.

Step 2 Heneicosasodic salt of benzene-1, 3, 5-tricarboxylic acid Tris. { [2-chloro-5- (hepta-O-sulfate-β-D-lactosyloxymethyl) phenyl] amide} A solution of 126 mg (0.079 mmol) of tris benzene-1,3,5-tricarboxylic acid is stirred at 70 ° C for 2.5 days. { [2-chloro-5- (β-D-lactosyloxymethyl) phenyl] amide} and sulfur trioxide trimethylamine complex (1.5 g, 8.26 mmol) in DMF (20 mL). The reaction mixture is cooled rapidly, at room temperature, with H2O and concentrated in vacuo. The residue is purified by chromatography with Sephadex G-10 (elution with H2O). Cation exchange is effected using a Dowex 50 x 8 strongly acid resin column (of the formula Na) to provide 207 mg (70% yield) of the title compound as a tan solid, m.p. > 171 ° C (with decomposition): Partial H NMR (D 2 O, 400 MHz) d 8.73 (s, 3 H), 7.68 (s, 3 H), 7.67 (d, 3 H), 7.58 (d, 3 H), 5.13 (d, 3 H), 5.08 (d, 3 H), 5.01 (d, 3 H), 4.43 (t, 3 H), and 4.24 ppm (t, 3 H); 13 C NMR (D20; 100 MHz) d 168.1, 137.0, 134.7, 132.8, 130.3, 130.1, 130.0, 129.6, 128.5, 127.5, 100.9, 99.2, 77.6, 77.0, 75.6, 75.3, 75.04, 74.97, 73.2, 71.7, 70.0 , 66.5, and 66.3 ppm; mass spectrum (electrorored) (m-zNa) / z (M-4Na) 4_ and 725.9 (M-5 Na) 5"Analysis calculated for C66H63Cl3N3Na2i? 99S2l.21 H20: C, 19.23; H, 2.57; N, 1.02; S, 16.33, Found: C, 19.51; H, 2.61; N, 1.11; S, 15.97.

EXAMPLE 13 Step 1 Benzene-1, 3, 5-tricarboxylic acid Tris. { [3,5- bis (tetra-O-acetyl-b-D-glucopyranosyloxymethyl) phenyl] amide} To a solution of 861 mg (1.06 mmol) of 3,5-bis (tetra-O-acetyl-β-D-glucosyloxymethyl) phenylamine in THF (20 mL) containing 147 μL (1.06 mmol) of triethylamine, 93.6 are added. mg (0.353 mmol) of benzene-1,3,5-tricarboxylic acid chloride. The reaction mixture is stirred for 90 minutes, quenched with MeOH, diluted with CH2Cl, and washed with H2O. The organic phase is dried (MgSO 4) and concentrated to a yellow solid. Purification is achieved by trituration with ether, to provide 849 mg (0.327 mmol) of the title compound as colorless crystals, m.p. 120-129 ° C: XH NMR (CDCI3; 400 MHz) d 9.1 (broad s, 3 H), 8.73 (s, 3 H), 7.67 (s, 6 H), 6.99 (s, 3 H), 5.18 (t, 6 H), 5.10 (t, 6 H), 5.03 (dd, 6 H), 4.84 (d, 6 H), 4.65 (d, 6 H), 4.61 (d, 6 H), 4.27 (broad s, 12 H), 3.72 - 3.74 (m, 6 H), 2. 06 (s, 18 H), 2.01 (s, 36 H), and 1.97 ppm (s, 18 H); 13 C NMR (CDCl 3, 100 MHz) d 171.2, 170.2, 169.4, 163.9, 138.4, 138.2, 135.2, 129.2, 123.1, 119.5, 100.1, 72.8, 71.9, 71.3, 71.1, 68.4, 62.0, 20.8, 20.64, and 20.56 ppm . Analysis calculated for Cn7Hi4iN3? 63 2H2O: C, 53.36; H, 5.55; N, 1.60. Found: C, 53.29; H, 5.29; N, 1.61.

Step 2 Tetracoso-sodium salt of benzene-1, 3, 5-tricarboxylic acid Tris. { [3,5-bis- (tetra-O-sulfate-β-D-glucosyloxymethyl) phenyl] amide} A solution of 780 mg (0.300 mmol) of tris benzene-1,3,5-tricarboxylic acid is stirred at 50 ° C for 5 hours. { [3,5-bis (tetra-O-sulfate-β-D-glucopyranosyloxymethyl) phenyl] amide} in MeOH (15 mL) containing 8.1 mL (8.1 mmol) of 1 N NaOH. The reaction mixture is cooled to room temperature and quenched with 1 N HCl (7.2 mL, 7.2 mmol). The product, benzene-1, 3, 5-tricarboxylic acid tris. { [3,5-bis- (β-D-glucosyloxymethyl) phenyl] amide} , meets (338 mg, 81% yield) and 1 is used directly in the next reaction: partial H NMR (DMSO-d6; 400 MHz) d 10.73 (s, 3 H), 8.74 (s, 3 H), 7. 78 (s, 6 H), 7.22 (s, 3 H), 4.88 (d, 6 H), 4.57 (d, 6) H), 4.29 (d, 6 H), 3.71 (d, 6 H), and 3.48 (dd, 6 H); NMR 13C (DMSO-d6; 100 MHz) d 164.5, 138.7, 138.4, 135.3, 129.8, 123.0, 119.1, 102.2, 77.0, 76.8, 73.5, 70.1, 70.0, and 61.1 pp. A solution of 317 mg (0.200 mmol) of tris benzene-1,3,5-tricarboxylic acid is stirred at 70 ° C for 3 days. { [3, 5-bis- (β-D, glucosyloxymethyl) phenyl] amide} and trimethylamine sulfur trioxide complex (3.59 g, 25.8 mmol) in DMF (25 mL). The reaction mixture is cooled rapidly, at room temperature, with H2O and concentrated in vacuo. The residue is purified by chromatography with Sephadex G-10 (elution with H2O) to give a yellow solid which is discovered, by NMR, containing some residual trimethylammonium sulfate. Cation exchange is effected using a Dowex 50 x 8 strongly acidic resin column (of the Na form) to provide 771 mg (approximately 95% yield) of the title compound, m.p. 174 ° C, as a colorless solid: NMR * H (D20; 400 MHz) d 8.63 (s, 3 H), 7.68 (broad s, 6 H), 7.44 (broad s, 3 H), 5.02 (d, 6 H), 4.98 (d, 6 H), 4.85 (d, 6 H), 4.54 (t, 6 H), 4.46 - 4.50 (m, 12 H), 4.24 (dd, 6 H), and 4.16 ppm (dt, 6 H); 13 C NMR (D20; 100 MHz) d 167.7, 138.0, 137.1, 135.3, 129.9, 125.3, 122.0, 99.5, 76.2, 75.9, 73.4, 72, 6, 70.9, and 67.6 ppm. Analysis calculated for C69H69N3Na24 ???? S2. 24H20. 4 Na S0: C, 16.45; H, 2.34; N, 0.83; S, 17.82. Found: C, 16.51; H, 2.17; N, 0.84, S, 18.17.

EXAMPLE 14 Step 1 Benzene-1,3,5-tricarboxylic acid Tris. { [3,5- bis (ß-D-celobiosyloxymethyl) phenyl] amide} To a solution of 802 mg (0.577 mmol) of 3,5-bis- (hepta-O-acetyl-β-D-cellobiosyloxymethyl) phenylamine in THF (25 mL) containing 80.6 μL (1.28 mmol) of triethylamine are added. 51.2 mg (0.193 mmol) of benzene-1,3,5-tricarboxylic acid chloride. The reaction mixture is stirred for 90 minutes, quenched with MeOH, diluted with CH2Cl2, and washed with H2O. The organic phase is dried (MgSO 4) and concentrated to a light yellow solid. Purification is achieved by trituration in CH2Cl2 / petroleum ether, to provide 777 mg (94% yield) of the product. A solution of 777 mg (0.180 mmol) of the compound, in MeOH (25 mL), is treated with 8.1 L (8.1 mmol) of 1 N NaOH. After stirring for 2.5 hours, at 50 ° C, the reaction mixture was stirred. cool rapidly to room temperature with 1 N HCl (7.6 L, 7.6 mmol). The solid is combined and purified by reverse phase chromatography (RP silica 60) using elution of MeOH / H20 (3: 7) then by chromatography with Sephadex G-10 (elution with H2O) to provide 262 mg (57% yield ) of the title compound as a colorless solid, mp > 171 ° C (with decomposition): 13 C NMR (D 20, 100 MHz) d 165.3, 138.4, 137.2, 134.0, 130.0, 123.9, 119.9, 102.6. 101.7, 78.7, 75.9, 75.5, 74.6, 74.2, 73.1, 72.8, 70.6, 69.4, 60.5, and 60.0 ppm.

Step 2 Tetratetracontasodic salt of benzene-1,3,3-ticarboxylic acid Tris. { [3,5-bis- (hepta-Q-sulfate-β-D-cellobiosyloxymethyl) phenyl] amide} A solution of 139 mg (0.0543 mmol) of tris benzene-1,3,5-tricarboxylic acid is stirred at 70 ° C for 3 days. { [3,5-bis- (β-D-lactosyloxymethyl) phenyl] amide} and sulfur trioxide complex triethylamine (1.68 g, 12.07 mmol) in DMF (25 mL). The reaction mixture is cooled rapidly, at room temperature, with H2O and concentrated in vacuo. The residue is purified by chromatography with Sephadex G-10 (elution with H2O). Cation exchange is effected using a Dowex 50 x 8 strongly acid resin column (of the Na form) to provide 317 mg (85% yield) of the title compound as a tan solid, m.p. > 180 ° C (with decomposition): Partial H NMR (D 2 O, 400 MHz) d 8.64 (s, 3 H), 7.67 (s, 6 H), 7.46 (s, 3 H), 5.02 (d, 6 H), 4.96 (d, 6 H), 4.64 (t, 6 H), and 3.88-4.01 (m, 12 H); 13 C NMR (D 20; 100 MHz) d 167.7, 138.0, 137.0, 135.3, 129.9, 125.1, 121.8. 99.8, 77.7, 77.5, 77.4, 77.0, 74.2, 73.5, 73.4, 72.9, 71.0, 67.6, 66.1 ppm; mass spectrum (electrorored) (m-zNa) / z 883.4 (m-8 Na) ~, 738.2 (m-9 Na) 9", 662.0 (m-10 Na) 10", 599.6 (m-11 Na) " Analysis calculated for C? O5HmN3Na 2? I95S 2 • 42H20: C, 16.59; H, 2.58; N, 0.55; S, 17.71. Found: C, 16.47; H, 2.86; N, 0.45; S, 14.72.

It is noted that in relation to this date, the best method known to the applicant to carry out the aforementioned invention, is that which is clear from the present description of the invention. Having described the invention as above, the content of the following is claimed as property:

Claims (13)

1. A compound of formula I I characterized in that each of R 1, R 2, R 3, and R 4 are, independently, H, S 0 3 M, or and > each oligosaccharide group contains from 1 to 3 sugar groups; M is lithium, sodium, potassium, or ammonium; n is 1 or 2; X is halogen, lower alkyl having 1 to 6 carbon atoms, or lower alkoxy having 1 to 6 carbon atoms; And it is carbonyl or sulfonyl; or a pharmaceutically acceptable salt thereof.

2. A compound according to claim 1, of formula I: I characterized in that each of R 1, R 2, R 3, and R 4 are, independently, H, SO 3 M, or and each oligosaccharide group contains 1 or 2 sugar groups; M is lithium, sodium, potassium, or ammonium; n is 1 or 2; X is halogen, lower alkyl having 1 to 6 carbon atoms, or lower alkoxy having 1 to 6 carbon atoms; And it is carbonyl; or a pharmaceutically acceptable salt thereof.

3. A compound according to claim 2, characterized in that it is the tris salt. { [2-methyl-5- (tetra-O-sulfate-β-glucopyranosyloxy-methyl) phenyl] amide} dodecasóica of benzene-1, 3-5-tricarboxylic acid, or a pharmaceutically acceptable salt thereof.

4. A compound according to claim 2, characterized in that it is the tris salt. { [2-methyl-5- (hepta-O-sulfate-β-cellobiosyloxy-ethyl) phenyl] -amidajheneicosodate of benzene-1, 3-5-tricarboxylic acid, or a pharmaceutically acceptable salt thereof.

5. A compound according to claim 2, characterized in that it is the tris salt. { [2-Chloro-5- (hepta-O-sulfate-β-D-maltosiloxy-methyl) phenyl] -amidajheneicosodate of benzene-1, 3-5-tricarboxylic acid, or a pharmaceutically acceptable salt thereof.

6. A compound according to claim 2, characterized in that it is the tris salt. { [2-Chloro-5- (hepta-O-sulfate-β-D-cellobiosiloxy-methyl) phenyl] -amidajheneicosodate of benzene-1, 3-5-tricarboxylic acid, or a pharmaceutically acceptable salt thereof.

7. A compound according to claim 2, characterized in that it is the tris salt. { [2-Chloro-5- (hepta-O-sulfate-β-D-lactosyloxymethyl) -phenyl] -amide-amino-sodium acid of 1,3-tricarboxylic acid, or a pharmaceutically acceptable salt thereof.

8. A compound according to claim 2, characterized in that it is the tris salt. { [3, 5- (tetra-O-sulfate-β-D-glucosyloxymethyl) -phenyl] amide} , tetracosodate of benzene-1,3,5-tricarboxylic acid, or a pharmaceutically acceptable salt thereof.

9. A compound according to claim 2, characterized in that it is the tris salt. { [3,5-bis- (hepta-O-sulfate-β-D-cellobiosyloxymethyl) -phenyl] amide} , tetratetraconsasodic acid of benzene-1,3,5-tricarboxylic acid, or a pharmaceutically acceptable salt thereof.

10. A method for the treatment of a human suffering from a condition characterized by excessive proliferation of smooth muscle, and the method is characterized in that it comprises administering to a human, an effective amount of the compound of the formula I wherein each of R 1, R 2, R 3, and R 4 are, independently, H, SO 3 M, or and each oligosaccharide group contains from 1 to 3 sugar groups; M is lithium, sodium, potassium, or ammonium; n is 1 or 2; X is halogen, lower alkyl having 1 to 6 carbon atoms, or lower alkoxy having 1 to 6 carbon atoms; And it is carbonyl or sulfonyl; or a pharmaceutically acceptable salt thereof.

11. A pharmaceutical composition, characterized in that it comprises an effective amount of a compound of formula I wherein each of R 1, R 2, R 3, and R 4 are, independently, H, SO 3 M, or and each oligosaccharide group contains from 1 to 3 sugar groups; M is lithium, sodium, potassium, or ammonium; n is 1 or 2; X is halogen, lower alkyl having 1 to 6 carbon atoms, or lower alkoxy having 1 to 6 carbon atoms; And it is carbonyl or sulfonyl; or a pharmaceutically acceptable salt thereof.

12. The use, to treat a human suffering from a condition that is characterized by excessive proliferation of smooth muscle, of an effective amount of the compound of formula I wherein each of R 1, R 2, R 3, and R 4 are, independently, H, SO 3 M, or and each oligosaccharide group contains from 1 to 3 sugar groups; M is lithium, sodium, potassium, or ammonium; n is 1 or 2; X is halogen, lower alkyl having 1 to 6 carbon atoms, or lower alkoxy having 1 to 6 carbon atoms; And it is carbonyl or sulfonyl; or a pharmaceutically acceptable salt thereof.

13. The use, in the manufacture of a medicament for the treatment of a human suffering from a condition that is characterized by excessive proliferation of the smooth muscle, of an effective amount of the compound of the formula I wherein each of R 1, R 2, R 3, and R 4 are, independently, H, SO 3 M, or and each oligosaccharide group contains from 1 to 3 sugar groups; M is lithium, sodium, potassium, or ammonium; n is 1 or 2; X is halogen, lower alkyl having 1 to 6 carbon atoms, or lower alkoxy having 1 to 6 carbon atoms; And it is carbonyl or sulfonyl; or a pharmaceutically acceptable salt thereof.