MXPA97007158A

MXPA97007158A - Fucopepti

Info

Publication number: MXPA97007158A
Application number: MXPA/A/1997/007158A
Authority: MX
Inventors: Wong Chihuey; Kajimoto Tetsuya
Original assignee: The Scripps Research Institute
Priority date: 1995-03-21
Filing date: 1997-09-19
Publication date: 1998-11-09

Abstract

The compounds of the formula I: wherein R is CH3, and R1 is a peptide residue, or R1 is OH and R is a peptide residue, has pharmacological activity as sialyl Lewis X mimics, for example in the prevention or treatment of disorders or diseases that are measured by the selectin link in cellulose adhesion

Description

FUCOPEPTIDES The present invention relates to fupeptides, to a process for their production, to their use as a pharmaceutical product, and to pharmaceutical preparations containing them. It is widely accepted that a family of receptors, selectins, are involved in the recognition of different circulating cells, by the endothelium and platelets, and have a role in certain diseases, including cancer, autoimmune disorders, inflammation, atherosclerosis, and blood coagulation. . There are three known members of this family: L-selectin, P-selectin, and E-selectin. E-selectin (also referred to as endothelial leukocyte adhesion molecule, ELAM-1), is a cell surface protein inducibly expressed in endothelial cells. For example, its production increases in vascular endothelial cells when adjacent tissue has been damaged, or when it has been invaded by a microorganism. E-selectin recognizes sialyl Lewis X (SLex), which is a cell surface carbohydrate ligand found in neutrophils and monocytes anchored to its outer membrane by glycoproteins and / or integral membrane glycolipids. SLex mediates the binding of neutrophils and monocytes with activated vascular endothelial cells, through the linkage with E-selectin, so that these leukocytes can diffuse into damaged tissue. However, there are many situations in which the recruitment of leukocytes by adhesion to endothelial cells is abnormal and excessive, and the end result is tissue damage instead of repair. Although it has been considered that SLex is potentially useful as an anti-inflamatory agent, and its large-scale synthesis has been developed for its clinical evaluation, this natural saccharide can only be used as an injectable form in the cases presented with acute symptoms, since it is orally inactive and has a short half-life in the blood. Accordingly, there is a need for compounds that can interfere with the binding of ligands with selectins, and that impede the initial cell adhesion process. According to the invention, a compound of the formula I is provided: where: i) R is CH3, and either R? is a radical of the formulas (ai) or (a2) (a,) (a where: m is 2 or 3; n is 2 or 3; it is a cation; R2 is H or a saturated or unsaturated hydrocarbon residue with up to 20 carbon atoms, optionally bearing at the? a formyl group, or an acetal group of alcohol of 1 to 4 carbon atoms or diol acetal of 2 to 4 carbon atoms; R3 is H, -CH2OH, or -CH2CH2OH; and R is H, alkyl of 1 to 4 carbon atoms, CH2OH, -CH2CH2OH, or -CH2CH2CH2OH, with the proviso that: 1) one of R3 and R4 is H, and 2) when R4 is H, R3 is - CH2OH or -CH2CH2OH, and 3) when R3 is H, R "is CH3, -CU2OH, CH2CH2OH or -CH2CH2CH2OH; O well; Ri is a radical of formula (b): (b) where: is: MOOC - CH2 -hr- CH-CO- NH- C- (b,) P,, _.I rent (b2) (b3) MOOC- where: p is 1 or 2; q is 2 or 3; r is 1 or 2; R6 is H, NH2, or -NHRX, where R? it's a protective group of amino; R7a is -CH2OH, -CH2CH2OH, or -CH (OH) -CH2OH, and R7b4 is H, or each of R a and R7b is CH? OH; R11 is H, or -OH; Rn is - (CH2) j -COOM or -S03M, where j is 1, 2, or 3; Y M is as defined above; the second hydroxy substituent of the phenyl group being in (b4) at any meta position; or Ri is a radical of formula (c): I O-CH-C. I (c) R, - CO - NH - CH - CORβ where: Re is OMi, ORH, RS-RP, OR -NHRy, where Mi is a cation, R? is a saturated or unsaturated hydrocarbon residue, R3 is a spacer group, Rp is a phosphatidyl residue, and Ry is a saturated or unsaturated lipophilic residue; and R9 is: MOOC- 0-C, -4 to uyl (C3) (CJ where: s is 1 or 2, t is 1 or 2, v is 2 or 3, M, R6, Rllf and R13 are as defined above; and R10a is -CH2OH, -CH2CH2OH, or -CH (OH) -CH2OH, and R10b is H, or each of R10a and R10b is CH20H; the second hydroxy substituent of the phenyl group being in (c2) at any meta position; or where: (ii) R? is OH, and R is a radical of the formula (d): R R 12a I H i MC C-tCH 2 -t - CH - CO - N - C - CO - NH - C - (d) I R 12b wherein: w is 1 or 2; R12a is -CH (0H) - (CH2) X-0H, and R12b is H, or each of Ri2a and Ri2b is independently -CH2OH or -CH2CH2OH; x is 2 or 3; and Re and M are as defined above.

M can be H * or any salt-forming cation for a carboxy or sulfate which maintains the water solubility of the compound of formula I, for example, a monovalent cation or an equivalent of a polyvalent cation, for example an alkali metal ion , such as lithium, sodium, or potassium; an alkaline earth cation, such as calcium or magnesium; as well as zinc, iron, and aluminum ions, and the ammonium ion (NH + 4). M is preferably H +, Li +, or Na +. It is preferred that the cation M is a pharmaceutically acceptable cation. When the cation is polyvalent, an appropriate number of molecules of the formula I, or a mixture of compounds of the formula I, and one or more appropriate anions, such as acetate, chloride, carbonate, and the like are also present. Mi may independently have one of the meanings given above for M, preferably identical to M. When Rx is an amino protecting group, this group may be as disclosed in "Protective Groups in Organic Synthesis", T.W. Greene, J.Wiley & Sons NY, 2nd edition, chapter 7, 1991, and references thereto, preferably a pharmaceutically acceptable amino protecting group, particularly tertiary-carbonyl butoxy or benzyloxycarbonyl.

When R2 in the radical of the formula (ai) is a saturated or unsaturated hydrocarbon residue, it can be, for example, alkyl of 1 to 20 carbon atoms, alkenyl of 2 to 20 carbon atoms, or alkynyl of 2 to 20. carbon atoms. Examples of R2 that carry a formyl group include, for example, 2-oxo-ethyl, 3-oxo-propyl, 5-oxo-pent-3-enyl, 8-oxo-octyl, and 10-oxo-dec-4. -enilo. When R2 is an alkyl or alkenyl acetal group formed from an alcohol of 1 to 4 carbon atoms or a diol of 2 to 4 carbon atoms, it may be, for example, any of the acetals which can be prepared starting from the above aldehydes using alcohols of 1 to 4 carbon atoms, or diols of 2 to 4 carbon atoms, for example, methanol, ethanol, isopropanol, secondary butanol, normal butanol, ethylene glycol, propylene glycol, 2, 3-butanediol, 1,4-butanediol, or 1,3-butanediol. R 2 is preferably H. Any alkyl of 1 to 4 carbon atoms as R in the radical of the formula (ai), is preferably CH 3. When Rβ is Rs-Rp or NHRy, the resulting compound of formula I can be used, or suitably used in a liposomal preparation as part of the liposome membrane, as a means to administer this compound. The spacer group Rs is a residue linking the carbonyl with the oxygen of the phosphatidyl residue, for example, a hydrocarbon residue. The phosphatidyl residue is a glycerophosphate esterified with saturated and / or unsaturated fatty acids, for example myristic, palmitic, stearic, palmitoleic, or oleic acid. R y can be a saturated or unsaturated aliphatic residue optionally comprising or interrupted by a functional group, for example, -CO-, for example, a residue based on a dicarboxylic acid diester carrying saturated or unsaturated fatty aliphatic residues. Ry is preferably: wherein each of X and Y is independently alkyl of 8 to 20 carbon atoms, or alkenyl of 8 to 20 carbon atoms, and Z is a bridging group, for example a polyethyleneoxy group, preferably a poly (3) group -50, preferably 3-15) -ethyleneoxy, or? N -alkylene group of 1 to 4 carbon atoms-CO- or -phenylene-CO-. In the compounds of the formula I, the following meanings are preferred, either individually or in any combination or sub-combination: 1. R is CM3, and R? is a radical of the formula (ai). 2. In the radical of the formula (ai), R2 is H. 3. In the radical of the formula (ai), R3 is H and R4 is - CH20H. 4. In the radical of the formula (ai), R3 is H and R-? It's CH3. 5. R is CH3 and Ri is a radical of the formula (b). 6. In the radical of formula (b), R5 is a radical (bx). 7. In the radical (b-, R7a is CH2OH or CH (OH) -CH2OH, and R7b is H. 8. In the radicals (a2) and (b3), the -O-alkyl group of 1 to 4 carbon atoms, is preferably OCH3 9. R is CH3, and R1 is a radical of the formula (c) 10. In the radical of the formula (c), R8 is alkoxy of 1 to 6 carbon atoms, preferably methyl or ethyl 11. In the radical of formula (c), R9 is a radical (c- 12. In the radical { cx), R10a is CH (OH) -CH20H, and R10b is H. 13. In the radical of formula (c), R9 is a radical (c2). 14. In the radical (c2), R6 is NH2. 15. R is OH, and R is a radical of the formula (d). 16. In the radical of formula (d), R12a is CH (OH) - (CH2)? -OH, and R12b is H.

The compounds of the formula I may comprise one or more asymmetric carbon atoms. It will be understood that the present invention includes all individual isomeric forms, enantiomers, and diastereoisomers, as well as mixtures, e.g., racemates, unless otherwise reported.

In the radical of the formula (ax), the asymmetric carbon atom bearing R3 preferably has the following configuration: In the radical of the formula (bx), when the carbon atom carrying R7a and R7b is asymmetric (i.e., R7b is H), it preferably has the following configuration: * 7th H The same is applied to the asymmetric carbon atom carried by Rxoa 'when R10b is H, in the residue (C. In the radicals (b2) and (c3), the pyrrolidinyl moiety preferably has the following configuration: In the radicals (b) and (c2), the asymmetric carbon atom carrying the substituted benzyl fraction preferably has the following configuration: The radical of formula (c) preferably has the following stereochemistry: / COOR. R9 - CO - HN In the radical of formula (d), when R12b is H "the asymmetric carbon atom bearing R12a preferably has the following configuration: Ri2a ^ o > The present invention also includes a process for the production of the compounds of the formula I. These can be produced in analogy to known methods. The compounds of the formula I can be produced, for example, by the removal of at least one protecting group, which is present in a compound of the formula I in protected form, for example, the protected form of amino and / or hydroxy. The compounds of the formula I in protected form are mainly compounds in which the hydroxy groups present in the fucose fraction are protected. The groups that can be employed in the present invention to block or protect the hydroxy group are well known to those skilled in the art, and preferably these groups can be removed, if desired, by methods that do not result in destruction. appreciable of the remaining portion of the molecule, for example, by chemical or enzymatic hydrolysis, treatment with chemical reduction agents under moderate conditions, and ultraviolet light radiation, or catalytic hydrogenation. The hydroxy protecting groups (blockers) that are conveniently used are those that are common in carbohydrate chemistry, especially for primary alcohols, secondary alcohols, and vicinal cis and trans diols. Suitable hydroxy protecting groups can be for example, acyl groups, such as acetyl, trichloroacetyl, phenoxycarbonyl, benzyloxycarbonyl, benzhydryloxycarbonyl, trityloxycarbonyl, and 2,2,2-trichloroethoxycarbonyl; ether groups such as methoxybenzyl, benzyloxymethyl, allyl, benzyl, p-methoxybenzyl, p-nitrobenzyl, benzhydryl, trityl, or triorganosilyl groups, such as trialkyl (of the ß carbon atoms) silyl (for example, trimethylsilyl, triethylsilyl) , tri-isopropylsilyl, isopropyldimethylsilyl, tertiary butyl-dimethylsilyl, methyldi-isopropylsilyl, or tertiary-methyl-butyl-silyl), tertiary butyl-diphenylsilyl, triarylsilyl (for example, triphenylsilyl, tri-p-xylsilyl), or triaralkylsilyl (for example, tribenzylsilyl). Examples of these and other hydroxy protecting groups suitable, for example, for the protection of the 1,2- or 1,3-dihydroxy groups, for example the cyclic ether groups, such as methylene acetal or ethylidene acetal optionally substituted, and the methods for their formation and removal, are known in this field, for example, see Groups in Orsanic Svnthesis. second edition, T.W. Greene and P.G.M. Wuts, John Wiley &; Sons, New York, 1991, chapter 2, and references therein. The compounds of the formula I wherein R6 is NHR ?, can also be converted into compounds of the formula I wherein R6 is NH2, by removal of the amino protecting group Rx. The compounds of formula I thus obtained can be recovered in free form or in salt form. The compounds of the formula I in protected form, wherein R is CH3 and R? is a radical of formula (a), used as starting materials, can be produced, for example, as indicated in Scheme 1.

Scheme l derivative X is a hydroxy protecting group, for example, as indicated above. R 'is an leaving group, for example, an amino protecting group, for example Boc or Fmoc. Preferably, the starting materials are used as specific enantiomers in order to obtain the compounds of the formula I with the desired configuration.

The compounds of the formula I in protected form, wherein R is CH 3 and R x is a radical of the formula (b) used as starting materials, can be prepared, for example, as indicated in Scheme 2.

Scheme 2 Compound II coupling Ry is a leaving group, for example, halogen, preferably F. Step ii is a reduction which is intended to include well-known reduction methods for the azido group, such as reaction with a phosphine, for example, triphenyl phosphine, or a hydride, for example, lithium-aluminum hydride. Step iii is an amide linkage coupling, for example as is known in the peptide chemistry art. Compound II can be selected, for example, from Asp-Ser-OH, Glu-Ser-OH, Glu- (α-hydroxymethyl) Ser-OH in protected form, for example, Compound 10. Step iii also may comprise coupling with an appropriate amino acid in protected form, followed by reaction with a diacid derivative as disclosed in Scheme 1. Compounds of formula I in protected form, wherein R is CH3 and R? is a radical of formula (c), can be prepared, for example, as indicated in Scheme 3.

Scheme 3 Compound IV R8a is alkyl of 1 to 6 carbon atoms. Ra is an amino protecting group. Compound IV can be selected, for example, from 2-amino-3,4-dihydroxy-butyric acid, α-hydroxymethrin serine, or Glu-Tyr-OH in protected form, eg, (2S, 3R) acid -N-Boc-2-amino-4-benzyloxy-3-hydroxybutyric acid. RgaCO is the amino acid residue (optionally completed with a diacid residue) of Compound IV. Steps i) and ii) are amide bond coupling reactions carried out according to conventional procedures. Step ii) may also comprise coupling with an appropriate amino acid in a protected form, followed by reaction with a diacid derivative, for example, as disclosed in Scheme 1. Step ii) may also further comprise the conversion of -COOR8a to -C0R8, for example, into an acid, an acid salt, a lipophilic ester, or a lipophilic amide, for example, as disclosed in Examples 44 and 45. Compound III can be used in the form of one or the other of the individual enantiomers, or in the form of mixtures.

Scheme 3 above illustrates the preparation of a compound of formula I, wherein R is CH3, and R? is a radical of formula (c), with the preferred configuration. The compounds of the formula I in protected form, wherein R? is OH and R is a radical of the formula (d), wherein R? a is CH (OH) - (CH2) x-OH, used as starting materials, can be produced, for example, as indicated in Scheme 4.

Scheme 4 Coupling Preferably, vicinal groups (hydroxy protecting groups) together form in the fucose fraction. The above reactions can be carried out in analogy with known methods, for example, as described in the following examples. When the production of the starting materials is not particularly described, the compounds are known or can be prepared in a manner analogous to methods known and practiced in the art. The following examples are illustrative of the invention. All temperatures are in ° C. The following abbreviations are used: Ac = -COCH3 Bn = benzyl Boc = tertiary butoxycarbonyl Fmoc 9-fluorenylmethoxycarbonyl DAST trifluoride diethylaminosulfur EDCI hydrochloride carbodi-i ida 1- (3-dimethylaminopropyl) 3-ethyl EDAC carbodi-imide l-ethyl-3- (3-dimethylaminopropyl) DMF = dimethyl formamide HOBT 1-hydroxybenzotriazole TFA = trifluoroacetic acid Example 1: Compound 6 is deprotected by treatment with hydrogen gas over a palladium / carbon catalyst, followed by treatment with sodium methoxide, to give the sodium salt of Compound 7.

? NMR (500 MHz, D 2 O) d 1.16 (d, J = 6.3 Hz, 3 H), 1.75-1.92 (m, 4H), 2.35 (m, 1 H), 3.20 (m, 1 H), 3.43 (m, ÍH), 3.55 (dd, J = 6.0, 12.0 Hz, 1 H), 3.67 (dd, J = 3.0, 12.0 Hz, 1 H), 3.73 (m, 2 H), 2.8 4.02 (m, 4 H); , 3 C NMR (125 MHz, D 2 O) d 16.6, 21.6, 24.5, 34.3, 35.4, 37.5, 56.5, 63.4, 68.2, 68.7, 70.8, 71.9, 72.6, 74.4, 166.8, 172.5, 177.0; electrospray mass m / z 423 [(MH) +; calculated for 17H31 ° 10N2: 23] - Compound 6, used as a starting material, can be prepared as follows: a) fucose tetra-acetate is treated with allytrimethylsilane and boron trifluoride stearate in dry acetonitrile at room temperature to give Compound 2 (the ratio a: β is greater than 10: 1). Kozikowsky, A.P. and Sorgi, K.L. Tetrahedron Lett. (1983), 24: 1563. b) Compound 2 is ozonolized by its reaction with ozone in the presence of triphenyl phosphine. The aldehyde product is then subjected to reductive amination by treating it with hydrogen gas on a palladium / carbon catalyst, in the presence of ammonium acetate, to give the corresponding fucose tetra-acetate carrying a group of 2 -aminoethyl (Compound 3). c) Compound 3 obtained above, is coupled with (lS, 2R) -2-N-Boc-amino-4-benzyloxy-3-hydroxybutyric acid (prepared from glycine and O-benzylglycolic aldehyde by a reaction catalyzed by aldolase of threonine according to Wong et al., Tetrahedron Lett 1995, 36. 4081), in a solution of l- (3-dimethylaminopropyl) -3-ethyl carbodiimide hydrochloride, to form the amide of Compound 5: d) The protecting group of Compound 5 is removed by its treatment with ethyl acetate in an acid solution (4N HCl). Then the deprotected group is treated on site with glutaric anhydride (in a pH regulator of triethyl amine) to obtain Compound 6. Following a procedure analogous to that of Example 1 above, compounds of the formula can be prepared: where R1; R2, n and m have the meanings indicated in Table 1 below.

Table l Ex. R, m 5c CH2CH2OH H 3 Following a procedure analogous to that of Example 1, but using the appropriate starting materials, the compound of Example 6 can be obtained: Example 7: 11 12 Compound 11 (122 milligrams, 0.12 millimoles) is dissolved in methanol (2 milliliters), Pd (OH) 2 on carbon (20 milligrams) is added, and the mixture is stirred under hydrogen (1 atmosphere) for 12 hours. The catalyst is filtered through Celite, and the product is purified by chromatography on silica gel (CHCl3 / methanol, 3: 1), and chromatography on P2 biogel (H20). Compound 12 is obtained after purification.

«H NMR (500 MHz, D2O) 61.48 (d, 3H, J = 7.0 Hz, H-6), 1.47-1.74 (m, 4H), 1.74 (s, 9H), 2.00-2.11 (m, 3H), 2.47 (m, ÍH), 3.64 (s, ÍH, H-4), 3.76 (m. ÍH), 3.97-4.25 (m, 6H, H-2, H-3, H-5), 4.65 ^ .74 (m, 2H), 5.31 (d, ÍH, J = 3.5 Hz, Hl). l3C NMR (125 MHz, D2O) d 15.80, 23.73, 24.51, 28.18, 30.18, 31.60, 39.68, 49.47, 53.42, 55.89, 55.99, 62.39, 67.12, 68.42, 70.08, 72.39, 94.38, 94.65, 170.9, 171.1, 174.7 , 178. 1. HRMS calculated for C24H41N3012Cs (M + CS +) 696. 1745, found 696.1717. Compound 11, used as a starting material, can be prep 8, 2,4-tri-O-benzyl-L-fucopyranose is dissolved in anhydrous dichloromethane at 0 ° C, and diethylaminosulfur trifluoride (1.06 grams, 0.0066 moles) is added dropwise. The mixture is then stirred at 0 ° C for 30 minutes, and the reaction is quenched by the addition of water. The aqueous solution is extracted with dichloromethane, and the organic fractions are combined, dried over MgSO4, and filtered. The solvent is evaporated, and fluoride is used for glycosylation without further purification. Molecular sieves of 4 A are added, tin (II) chloride (1.67 grams, 0.0088 moles), and silver perchloride (1.82 grams, 0.0088 moles), to a solution of fluoride in anhydrous dichloromethane (20 milliliters) at 0 ° C. The mixture is stirred for 5 minutes, and (R, R) azidocyclohexanol (0.93 grams, 0.0066 moles) is added. The reaction is warmed to room temperature, and stirred for 6 hours. After filtration through Celite, the filtrate is concentrated and applied to chromatography on silica gel (hexane / ethyl acetate, 8: 1). Compound 8 is obtained as a clear oil.

Compound 8 (1.3 grams, 2.33 millimoles) is dissolved in tetrahydrofuran (10 milliliters, containing 1 percent H20), and triphenyl phosphine (638 milligrams, 2.56 millimoles) is added. The mixture is stirred at room temperature for 5 hours. After evaporation of the solvent, the residue is purified by chromatography on a column of silica gel with CHCla / methanol (50: 1 -> 30: 1). Compound 9 is obtained as a syrup.

BnOOC c) O-Benzyl-N-Boc-L-aspartic acid is dissolved in anhydrous dichloromethane, and carbodiimide l-ethyl-3- (3-dimethylaminopropyl) (640.3 milligrams, 3.34 mmol) and N-hydroxysuccinimide ( 384.4 milligrams, 3.34 millimoles). The mixture is stirred at 4 ° C for 12 hours, and the solvent is evaporated. N-hydroxysuccinimide O-benzyl-N-aspartic ester is obtained by chromatography on silica gel (ethyl acetate / hexane, 1: 1.5 -> 1: 1). The N-succinimide O-benzyl-N-aspartic ester (603 milligrams, 1.44 millimoles) and O-benzylic serine (281 milligrams, 1.44 millimoles), are dissolved in dimethyl formamide (2 milliliters), and Et3N (1 milliliter) is added. ). The mixture is stirred at room temperature for 1 hour. After evaporation of the solvent and chromatography on silica gel (CHCl3), product 10 is obtained. D) Acid 10 (141 milligrams, 0.29 mmol), carbodiimide l-ethyl-3- (3-dimethylaminopropyl) ) (81 milligrams, 0.42 millimoles), and 1-hydroxybenzotriazole (57 milligrams, 0.42 millimoles), are dissolved in dichloromethane (2 milliliters) at room temperature, and stirred for 5 minutes, before Compound 9 is added ( 148 milligrams, 0.56 millimoles). The mixture is stirred at room temperature for 3 hours, and the solvent is evaporated. The residue is applied to a column of silica gel. { hexane / ethyl acetate, 1.5: -1: 1), and Compound 11 is obtained as a syrup. 6. 5 Hz, H-6), 1.08-1.60 (m, 4H), 1.44 (s, 9H), 1.73 (m, ÍH), 1.94 (, 3H), 2.71 (m, ÍH), 3.05 (m, ÍH) , 3.65 (dd, ÍH, J = 3.5, 9.5 Hz), 3.76 (m, ÍH, H-4), 3.89 (dd, 1 H, J = 3.0, 9.5 Hz), 3.98 (dd, ÍH, J = 2.5 , 10.0 Hz. H-3), 4.04 (dd, ÍH, J = 3.5, 10.0 Hz, H-2), 4.10 (m, ÍH, H-5), 4.44 (m, ÍH), 4.61 (m, ÍH ), 4.68 (m, HH), 4.95 (d, HH, J = 3.5 Hz, Hl), 4.45-5.17 (m, 10H), 5.68 (m, HH), 7.33 (m, 25H). 13 C NMR (125 MHz, CDC13) d 16.72, 23.34, 23.82, 28.20, 36.29, 36.59, 50.42, 50.65, 52.45, 52.58, 52.82, 66.69, 66.72, 66.78, 69.26, 73.09, 73.18, 74.80, 75.98, 75.98, 77.73 , 79.35, 94.52, 128.0 (m), 135.2, 135.3. 137.5, 183.6, 139.0, 155.3, 169.6, 170.2, 170. 6, 171. 9. HRMS calculated for C59H71N3012Cs (M + CS +) 1146.4092, found 1146.4035.

Following a procedure analogous to that of Example 7 above, but using the appropriate starting materials, the following compounds of the formula can be prepared: where R5 is as indicated in Table 2.

Table 2 Ex. HRMS Calculated Found HO. CH, OH CH [M + Cs * J 611.1217 611.1241 HOfC- CH, - HOjC-f-CH, - .1241 CHtOH 0 H i 485.2111 485.2127 NßOjC-t-CH, - CO-N -CH- CHjOH OH ^ CH 1 H I [M * Cs *] HOiC-f-CH, - • CO-N -CH- 625.1373 625.1355 Table 2 (continued) E3 • Rs HRMS Calculated Found CH.OH _ ^ CH 12 H T [M + CV] HO, C - CH, - CH - CO • N - CH - 726.1850 726.1832 1 NHToc CH2OH V, OH "CH 3 H? NaOjC-CHj CH-CO-N-CH- * [M + Na *] 1 516.2169 516.2169 NH.

HO, C-f-CH1 + -CO Table 2 (continued) Ex. R5 Calculated HRMS Found co-npueßto of Example 11: 'H IRMN (500 MHz. D2O) d 1.10 (d, 3H, J = 6.5 Hz, H-6), 1.20 (m.4H). 1.75 (m, 6H). 2.16 (m.H.) 2.56 (t.2H, J = 7.5 Hz), 2.33 (m.2H), 3.41 (m.1H). 3.43 (m, 2H). 3.63 (m.2H, H-2 and, H-3), 3.69 (, 1H, H-4), 3.78 (m, 1H, H-5), 4.09 (ddd, 1H, J * 2.5, 6.0, 8.5 Hz), 4.46 (d., 1H, J, 2.5 Hz). 5.02 (d, 1H, J s 3.0 Hz, H-1). ? ac NMR (125 MHz, D20) d 15.85, 21.96, 23.71, 24.60, 28.84, 31.41, 35.38, 35.45, 53.67, 55.19, 62.73. 67.13. 68.26. 69.88. 71.65. 72.22. 75.19. 93.31, 172.3, 176.6.

Compound of Example 12: 1 H NMR. (500 MHZ, D20) d 1.12 (d, 3H, J = 6.5 Hz, H-6) .1.08-1.26 (m.4H), 1.39 (s.9H), 1.65-1.80 (m.3H), 2.14 (, 1H) .2.74 (m, 2H), 3.39-3.88 (m, 9H) .4.37 (m, 2H), 4.98 (d. 1H, J * 3.5 Hz, H-1). 13C NMR (125 MHz, D2O) d 15.82, 23.70, 24.58, 28.03, 29.30, 31.78, 37.32.52.05, 53.40.55.55.62.93.67.14.68.32, 69.95, 71.99, 72.25, 76.23, 82.21.94.06. 157.6.170.8, 173.4, 175.8.

Compound of Example 13: iH NMR (500 MHZ, D2O) d 1. 15 (d.3H.J - 6.5 Hz. H-6), 1.10-1.40 (m, 4H) .1.66-1.85 (m.3H) .2.16 (, 1H), 2.59 (dd.1H, J = 8.5. 17.5 Hz) .2.72 (dd.1H.J = 5.0.17.5 Hz), 3.40 (m.1H), 3.51 (dd, 1H.J = 6.0, 12.0Hz), 3.56 (dd.1H, J ± 3.5, 10.0 Hz, H-3) .3.60 (m, 1H), 3.64 (dd, 1Ht J = 4.0, 10.0 Hz, H-2) .3.69 (m, 1H, H-4), 3.73 (m, 1H) .3.85 (m.1H) .3.91 (m.1H.H-5) .4.15 (m, 1H), 4.40 (d, 1H, J * 7.0 Hz), 5.00 (d.1H.J = 4.0 Hz. H-1). "C NMR (125 MHz, D20) d 15.77.23.69.24.62.29.06.31.80, 38.41.51.47.53.40.55.59.62.76, 67.09.68.35.69.91.71.79.72.24.75.68, 93.63, 170.6.176.8, 176.8.

Example 18; The benzyl groups of 15 are dissociated by hydrogenation according to the procedure of Example 5. Compound 16 (amorphous): XH NMR (400 MHz, D20) 5 4.98 (d, J = 3.2 Hz, 1 H, Hl), 4.61 ( d, J = 2.2 Hz, 1 H), 4.56 (d, J = 7.2 Hz, 1 H), 4.46-4.44 (dd, J = 2.2 and 6.4 Hz, 1 H), 4.28-4.13 (m, 3H), 4.02-3.98 (m, ÍH), 3.86-3.59 (m, 5 H), 2.42-2.22 (m, 4 H), 1.90-1.79 (m, 2H), 1.30-1.18 (m, 9H); negative ion mass by electrospray (deagglomeration potential = -80 V) m / z 523 [(M-H); calculated for C21H36N2013: 524]. Compound 15, used as a starting material, can be prepared as follows: a) BnO OBn - - »O ----? OBn First, L-fucose is converted to tribenzylsuclucosyl phosphite according to Wong et al., J. Org. Chem. 1994, 59, 864. The resulting compound (1.0 equivalent) is coupled with Boc-L-Thr-OEt (1.1 equivalents) using trifluoromethanesulfonic acid (1.0 equivalents) as a catalyst in methylene chloride at 0 ° C, to give the Compound 13 after a conventional work and purification. 14 First deprotection of Boc 13 is made in 30 percent trifluoroacetic acid in 0.1 M CH2C12 at 25 ° C, 30 minutes; it is quenched with water, washed with NaHCO 3, dried with sodium sulfate; and purification by chromatography by evaporation gives the corresponding free amine. This amine (1.0 equivalent) is coupled with 1.1 equivalents of (2S, 3R) -N-Boc-2-amino-4-benzyloxy-3-hydroxybutyric acid using 1.5 equivalents of carbodiimide hydrochloride l- (3-dimethylaminopropyl) -3-ethyl, 1.5 equivalents of HOBt, CH2C12 0: 1M, 0 ° C, 30 hours, to provide compound 14 after of normal work and conventional purification conditions by column chromatography by evaporation. c) Deprotection of Boc from compound 14 in 30 percent trifluoroacetic acid, 0.1 M CH2C12 at 25 ° C, 30 minutes, it is followed by quenching with water, washing with NaHCO3, and drying over sodium sulfate. Purification by evaporation chromatography is followed by coupling with 1.1 equivalents of monobenzyl glutarate, 1.5 equivalents of carbodiimide hydrochloride 1- (3-dimethylaminopropyl) -3-ethyl, 1.5 equivalents of HOBt, CH2C12, 25 ° C, 20 hours, for provide Compound 15. Compound 15: * H NMR (400 MHz, CDC13) d 8.00 (d, J = 9.1 Hz, 1 H), 7.21-7.50 (m, 25H). 6.75 (d, J = 7.6 Hz. 1 H), 5.12-4.57 (m.1 1 H), 4.35-3.99 (m, 7H), 3.80-3.60 (m, 5 H). 2.45 (t, J = 7.5 Hz, 2H). 2.43 (1. J = 7.0 Hz, 2H), 1.98 (t, J = 7.5 Hz, 2H), 1.20-1.26 (m, 6 H). 1.06 (d.J = 6.4 Hz, 3 H); HRMS for CJ6H-66N2O "+ Cs + (M + Cs +). cale 1 107.3614. found.1 107.3667.

Following a procedure analogous to that of Example 18 above, but using the appropriate starting materials, compounds of the formula can be prepared: wherein R8 and R9 are as defined in Table 3.

Table 3 Ex. Rß R * MS Calculated Found 9 H ^ ° ^ CH2OH ES (M-H) HO.C-f-CH, CO-NH-C-496 495 1 H \ H ES (M-H) HO, C-CH • CH, • CO-NH-C 543 * 42 T NH 2 Table 3 (continued) Ex. R, R, MS Calculated Found HO, C - (- CH, -) - CO HO, C- CH, -) - CO Table 3 (continued) Ex R. R, MS Calculated Found Example 29 19 20 A solution of glycopeptide 19 (23.7 milligrams, 40 micromoles) is stirred in 90 percent trifluoroacetic acid in water (1 milliliter) at room temperature. After 3 hours, the reaction solution is evaporated under reduced pressure, and azeotroped twice with toluene (2 x 5 milliliters). Nuclear magnetic resonance is performed on the crude product to ensure that the isopropylidene fractions are removed, and then the product is taken to the next step without purification. Then the crude glycopeptide is hydrogenated according to the procedure of Example 5, to give a white solid 20: Rf (4: 1: 1 nBuOH: H20: HOAc) 0. 29: XH NMR (400 MHz; D20) . 23 (d, J 3.7, Hlß), 4.55 (dd, J 7.9 and 4.7, Hlp), 4.37-4.32 (m, H2 '(a + P)), 4.15-4.05 (m, H5ß + H3 P), 3.95 -3.92 (m, H4β + H3'J, 3.86 (d, J 3.4, H4p), 3.83 (dd, J 10.3 and 3.2 H3a), 3.78 (dd, J 10.3 and 3.7, H2J, 3.74-3.68 (m, H5p ) 3.64-3.61 (m, H3p + C6? "ß + P)), 3.55-3.44 (m, H2" + COH, ^ "+ P)), 3.42-3.31 (m, CoH.H * ,, + P )), 2.62-2.48 (m, 02 '^ + P)) + C3"H? A + P)). 1.76-1.47 (m, C4? J (ß + P)) + C5? 2 (ß + P )); 13C NMR (100 MHz; D2O) 182.93, 179.11, 175.20, 174.66, 98. 86, 94.80, 75.19. 75.08, 75.02, 74.21, 72.91, 71.78, 71.48, 71.28, 71.18, 70.71, 70.52, 63. 84, 63.80, 61.00. 60.90, 60.48, 42.15, 42.03, 34.71, 34.19. 34.12, 31.84, 31.44, 31.14; High resolution mass spectrum (added with Nal): Found N + Na, 447.1570. C16H28N2011 requires M + Na 447.1591.

Compound 19, used as a starting material, can be prepared as follows: 17 carbodiimide hydrochloride l- (3-dimethylaminopropyl) -3-ethyl (95.4 milligrams, 500 micromoles) is added to a stirred solution of 6-amino-6-deoxy-1,2,3,4-diisopropylidene- cx-L-galactopyranoside (130 milligrams, 500 micromoles), (2S, 3RS) -2-N-Boc-amino-6-benzyloxy-3-hydroxyhexanoic acid (177 milligrams, 500 micromoles), 1-hydroxybenzotriazole (68 milligrams, 500 micromoles), and 4-methyl morpholine (108 microliters, 1,000 micromoles) in dry dimethyl formamide (5 milliliters) under argon at -20 ° C. The resulting mixture is stirred -20 ° C for 1 hour, and then let it slowly warm to room temperature. After 14 hours, the reaction solution is quenched with a 5 percent weight / volume citric acid solution (20 milliliters), and extracted with ethyl acetate (6 x 25 milliliters). The combined organic extracts are washed with a saturated solution of sodium hydrogen carbonate (50 milliliters), and a saturated solution of sodium chloride (50 milliliters), dried (t-gS04), and evaporated under reduced pressure. The residual oil is purified by flash chromatography (silica gel, using an elution gradient of 40 percent - * 50 percent - »66 percent ethyl acetate in hexane) to give compound 17 as a foam pale yellow: Rf (75 percent ethyl acetate in hexane) 0.66. 18 A glycopeptide solution (17) (59.5 milligrams, 100 micromoles) in 15 percent volume / volume trifluoroacetic acid in 4- (dicyanomethylene) -2-methyl-6- (4-dimethylamino) steryl-4-H-pyran Dry (1 milliliter), stir under argon at room temperature. After 2 hours, the solution is evaporated under reduced pressure, and the residual oil is dissolved in normal butanol: water: methanol (5: 3: 2) (10 milliliters). To the solution is added Dowex (Cl ", previously washed with methanol, 100 milligrams), and the mixture is stirred for 30 minutes, then filtered, and the solid is washed with methanol (3 x 5 milliliters), and the filtrate is filtered. and the combined washings are evaporated under reduced pressure.The residual oil is purified by flash chromatography (silica gel, using a dilution gradient of 19: 0.9: 0.1 - >).; 9: 0.9: 0.1 DCM: MeOH: NH 3 (ac)) to give compound 18 as a light brown oil: Rf (9: 0.9: 0.1 DCM: MeOH: NH 3 (ao) 0.42) c) Succinic anhydride is added (6.0 milligrams, 60 micromoles) to a stirred solution of aminoglycopeptide 18 (27.7 milligrams, 56 micromoles) in methanol (1 milliliter) at room temperature. After 1 hour, the solution is evaporated under reduced pressure. The residual solid is purified by flash chromatography (silica gel, using an eluate gradient of 5 -> 10 percent ethyl acetate in ethyl acetate), to give Compound 19 as a pale yellow gum. Rf (10% acetic acid in ethyl acetate) 0.49: IR (Film) cm-1 3303, 2980, 2935, 1644, 1558, 1436, 1382, 1255, .1211, 1167, 1109.1070.1006.901: 'H NMR (400 MHz, CDjOD) 7.32-7.23 (5H, m, aromatic), 5.44 (1H.D. J 5.0.2 x Hl) .4.59 (1H. dd.J 7.9 and 2.1, 2 x H3), 4.48 (2H. , s, 2 x CH2Ph) .4.38 ^ .34 (IH .2 x H7), 4.31 (IH dd, J 5.0 and 2.4, 2 x 112) 4.21 (III.D. J 7.9.2 x 114). 4.00-3.89 (1.5H.m.2 x 115 + 113 '). 3.84-3.76 (0.5H.m. H3 ') 3.52-3.46 (3H.m.2 x C6H, Hb + 2 x C6H2), 3.27-3.20 (ÍH.m.2 x C6 H. Hb) .2.56-2.48 (4H.m.2 x C2"H2 + 2 x C3" H2), 1.80-1.54 (4H, m, 2 x C4'H2 + 2 x C5'H2), 1.45 (3H. S.acetonidcMc) .1.40 (3H, sscetópido Me). 1. 32 (3H, spcid Me), 1.29 (3H.s.acetonidec), 3C NMR (100 MHz, CD3OD) 137.21, 126.78.126.24, 126.03.107.86.107.34.95.19.71.24.70.06, 69.56 , 69.28, 69.24, 68.58, 64.68, 56.66.38.02.28.96.28.35.24.54.23.83, 23.75, 22.62, 21.99; High resolution mass spectrum (added with Csl): Found M + Cs, 727.1875. C "H" N, On requires M + Cs, 727.1843.

Following a procedure analogous to that of Example 29, but using the appropriate starting materials, compounds of the formula can be prepared: where RA and RB are as defined in Table 4.

Table 4 Ex NHt 0 H OH HO, C - CH 2 - CH - CO - N - CH • CH - f - CH 2) - OH HO * 31 HOjC CHj • CO-NH-CH- OH CH CH, OH HOV 32 HO-C-Y- CH2- ^ CO-NH-CH-OH CH- CH ^ OH 33 H02C -f- CH, - + CO-NH-CH - w * OH CH- CHt -) - OH HO HOjC-t-CH ^ CO-NH Table 4 (continued) Ex 36 HO ^ CH, OH CH OH CH- 7 HO ^ CHjOH CH I ***** OH CH- HO.C-CH.-CH-CO-N I NH 8 HO ^ ^ .CHjOH CH * # OH I CH- H02C- (CH, - • CO-N H 9 (CHt) j-OH CH ***** OH I CH HO.C-CH.CH-CO-N i H NH, Table 4 (continued) E j. RA RB 0 1 2 HOjC-fCH ^ 3 CH, OH OH HOCH, CH. v-C V- Example 44 27: R "= Bn 28: R '= H To the solution of Compound 27 (91.4 milligrams, 59.2 micromoles) in MeOH (5 milliliters), Pd (0H) 2 on carbon (20 milligrams) is added. The mixture is stirred for 2 days under a hydrogen balloon. The catalyst is filtered, and the filtrate is evaporated in vacuo. The residue is applied to silica gel column chromatography (CHCl3: MeOH = 1: 1), and then to Sephadex LH20 gel filtration chromatography (eluted with MeOH), to obtain the desired amphiphile 28.

? NMR: (500 MHz, CD3OD) d 0.80 (6H.t, J = 6.5), 1.16 (6H, d, J = 5.5), 1.23-1.41 (52H, m), 1.60-1.71 (4H.m), 2.04 -2.17 (2H, m), 2.31-2.60 (6H, m), 2.66-2.77 (2H.m). 2.97-3.09 (2H.m) .3.31-3.40 (3H.m), 3.50-3.71 (14H.m) .3.79-3.87 (ÍH, m) .3.88-3.98 (ÍH, m), 4.04-4.26 (4H, m), 4.44-4.56 (3H, m), 4.95 (1H, brs). HRMS calculated for C62H "4N4O, 7Cs (M + Cs) 1319.7233, found 1319. 7264.

Compound 27, used as a starting material, can be prepared as follows: a) Compound 21 A suspension of dibenzyl-2,3,4-0-tribenzyl-aL-fucopyranosyl phosphite (138 milligrams, 0.204 mmol), threonine N-9-fluorenylmethoxycarbonyl allyl ester (77.5 milligrams, 1 equivalent), and molecular sieve of 4 A (470 milligrams), is stirred for 18 hours at room temperature.

To the mixture is added a solution of TMSOTf (13.6 milligrams, 0.3 equivalents) in CH2C12 (1 milliliter) at -15 ° C. After stirring for 2 hours at the same temperature, saturated NaHC03ac is added to quench, and the mixture is diluted with CH2C12. The organic layer is separated, dried over MgSO 4, and evaporated in vacuo.

The residue is applied to silica gel column chromatography (Hex: EtOAc = 4: 1) to obtain Compound 21.

Compound 22 To a solution of Compound 21 (104 milligrams, 0.13 mmol) in CH2C12 (5 milliliters), Et2NH (0.72 milliliters, 53 equivalents) is added, and the mixture is stirred for 18 hours at room temperature. The solvent and reagent are removed in vacuo, and the residue is purified by silica gel column chromatography (CH2C12 ~ CH2Cl2: MeOH = 100: 3) to obtain compound 22.

Compound 23 To a solution of Compound 22 (242 milligrams, 0.42 millimoles) and N-benzylic glutarylamide-L-proline (135 milligrams, 1 equivalent) in CHC12 (5 milliliters), 1-hydroxybenzotriazole is added successively (86 milligrams, 1.5 equivalents) and carbodi-imide l- (3-dimethylaminopropyl) -3-ethyl (105 milligrams, 1.3 equivalents) at 0 ° C. The reaction mixture is stirred for 20 minutes at 0 ° C, and the temperature is allowed to rise to room temperature within 18 hours. The reaction mixture is evaporated in vacuo, and the residue is dissolved with EtOAc. The EtOAc solution is washed with IN HCl, saturated NaHCO 3, and brine, successively. The organic layer is dried over MgSO 4 and evaporated in vacuo. The residue is applied to silica gel column chromatography (toluene-toluene: acetone = 2: 1), to obtain Compound 23.

Compound 24 To a solution of Compound 23 (169 milligrams, 0.192 millimoles) and morpholine (167 milligrams, 10 equivalents) in tetrahydrofuran (10 milliliters) and dimethyl formamide (1 milliliter), Pd (PPh3) 4 (23 milligrams, 0.1 equivalents) under an argon atmosphere at room temperature. After stirring for 24 hours, the reaction mixture is evaporated in vacuo. The residue is dissolved with EtOAc and washed with IN HCl. The organic layer is separated, dried over MgSO 4, and evaporated in vacuo. The residue is purified by chromatography on a silica gel column (CH2Cl2-CH2Cl2: MeOH 10: 1) to obtain Compound 24. b) Synthesis of the lipid part Compound 25 To a solution of oxalyl chloride (1.54 milliliters, 2N CH2C12 solution), add a solution of dimethyl sulfoxide (277 milligrams, 1.5 equivalents) in CH2C12 (3 milliliters), and a solution of 2- [2]. - (2-azidoethoxy) ethoxy] ethanol in CH2C12 (5 milliliters), each at a range of 2 minutes at -78 ° C. After the mixture is stirred for 30 minutes at the same temperature, Et ^ is added. 1 (1.64 milliliters) to the mixture at -78 ° C, and the temperature is allowed to rise to room temperature within 1 hour. The reaction mixture is poured into ice water and extracted with CHjCl 2. The organic layer is washed with IN HCl and saturated NaHCOj successively, dried over MgSOa, and evaporated in vacuo. The crude aldehyde is used without further purification. To a solution of the crude aldehyde and p-toluenesulfonic acid salt of 1 ', 3'-dicetyl-L-glutamate (500 milligrams, 1 equivalent) in tetrahydrofuran (3 milliliters) and MeOH (9 milliliters), NaCNBH3 is added ( 41 milligrams) at 0 ° C. The mixture is stirred at 0 ° C for 1 hour, and the temperature is allowed to rise to room temperature within 18 hours. The solvent is evaporated in vacuo, and the residue is dissolved with CH £12. After the solution is washed with IN HCl, the organic layer is dried over MgSO0, and evaporated in vacuo. The residue is applied to silica gel column chromatography (Hex-EtOAc = 4: 1) to obtain Compound 25.

Compound 26 To the solution of Compound 25 (320 milligrams, 0.427 mmol) in MeOH (13 milliliters), tetrahydrofurapo (3 milliliters), and IN HCl (1.3 milliliters), Pd on carbon (99 milligrams) is added. The mixture is stirred for 2 days under a hydrogen balloon. The catalyst is filtered, and the filtrate is evaporated in vacuo. Saturated NaHCO3 is added to the residue, and the mixture is extracted with EtOAc. The organic layer is dried over MgSO, and evaporated in vacuo to obtain Compound 26.

Coupling between sLex mimetic and lipid Comp this 27 To a solution of Compound 24 (94.6 milligrams, 0.113 mol) and Compound 26 (81.7 milligrams, 1 equivalent) in CH2C12 (5 milliliters), 1-hydroxybenzotriazole (23 milligrams, 1.5 equivalents) and carbodiimide 1- (3-dimethylamino-propyl) -3-ethyl (29 milligrams, 1.3 equivalents) successively at 0 ° C. The reaction mixture is stirred at 0 ° C, and the temperature is allowed to rise to room temperature within 20 hours. The reaction mixture is evaporated in vacuo, and the residue is dissolved with EtOAc. After the solution is washed with saturated NaHCO3, the organic layer is dried over MgSO4, and evaporated in vacuo. The residue is purified by column chromatography on silica gel (toluene: acetone = 1: 1), to obtain Compound 27. H NMR (500 MHz, CDC13) d 0.88 (6H, t, J = 7.0), 1.12 ( 3H, d, J = 6.0), 1.17 (3H, d, J = 6.0), 1.21-1.41 (52H,), 1.57-1.65 (4H, m), 1.88-1.97 (2H, m), 2. 12-2.20 (ÍH, m), 2.22-2.29 (ÍH, m), 2.38 (2H, t, J = 7.0), 2.45-2.83 (5H, m), 3.18-3.27 (2H, m), 3.42-3.59 (10H, m), 3.63 (1H, brs), 3.66 (1H, dd, J = 11.0, 4.0), 3.82 (IH, dd, J = 10.3, 3.0), 3.84 (IH, q, J = 7.0), 4.01-4.13 (4H, m), 4.48-4.58 (4H, m), 4.61 (ÍH, d, J = ll .5), .6 < 1H, d, J = 11.5), 4.69 (IH, d, J = 11.5), 4.74 (IH, d, J = 11.5), 4.75 (IH, d, J = 11.5), 4.92 (HH, d, J = 11.5), 4.96 (HH, d, J = 3.5), 5.06 (2H, s), 7.10 (HH, brt, J = 3.0), 7.25-7.43 (20H, m), 7.87 ( ÍH, t, J = 9.0), m / z C90H138N4 ° 17 1548 (M + H) • Example 45 Following a procedure analogous to that of Example 44, but using the appropriate starting materials, the following compound can be obtained: XH NMR (500 MHZ, CDC13), d 4.80 (d, J = 2.6 Hz, ÍH), 4.46 (t, J = 8.5 HZ, ÍH), 4.42-4.40 (m, 4H), 4.05-3.95 (m, 6H ), 3.76-3.43 (m, 6H), 2.23 (t, J = 7.7HZ, 2H), 2.30-2.20 (m, 3H), 2.12-2.01 (m, 2H), 1.98-1.91 (m, 4H), 1.85-1.77 (m, ÍH), 1.57-1.50 (m, 4H), 1.25-1.13 (bs.,> 50H), 1.11 (d, J = 6.3 Hz, 3H), 1.09 (d, J = 6.6 Hz , 3H), 0.78 (t, J = 6.7 Hz, 6H); MS m / e calculated for C59H106N4O16Cs (M + Cs +): 1259.6658, found 1259.6620. The compounds of formula I and their pharmaceutically acceptable salts exhibit pharmaceutical activity, and therefore, are useful as pharmaceuticals. In particular, they inhibit adhesion between the cells containing a selectin, such as E-selectin, on their surfaces, and the detector cells such as neutrophils or HL-60 cells having SLex on their cell surfaces, or a polysaccharide product. Synthetic SLex or a poly-SLea product. More particularly, the compounds of the formula I inhibit the binding of sLex with E-selectin, as indicated in the following test methods: a) Preparation of a soluble form of E-select.

The extracellular domain of E-selectin, fused to the constant region of the light chain (mCK) is cloned into the baculovirus release vector pVL941 (Invitrogen), and expressed in SF-9 cells. The soluble fusion protein is purified by affinity chromatography using the monoclonal antibody rat CK against mouse 187.1 coupled with Sepharose. Cell Link Assays. The compounds of formula I are tested for their ability to block adhesion of HL-60 cells to immobilized E-selectin on 96-well plates. Rat CK antibody against mouse is added to 96-well plates (20 micrograms / milliliter in carbonate / bicarbonate buffer, pH 9.5), and incubated overnight at 4 ° C. The plates are blocked with 3 percent bovine serum albumin in the assay regulator (20 mM HEPES, pH 7.4, containing 150 mM NaCl and 1 mM CaCl 2) for 8 hours at room temperature, and washed three times with the regulator of the trial. The mouse CK fusion protein of E-selectin (10 micrograms / milliliter in the assay regulator) is added, and incubated for 2 hours at 37 ° C, or overnight at 4 ° C. The plates are washed three times with the assay regulator. The compound to be tested is then added and pre-incubated for 15 to 30 minutes at 37 ° C. Transfer 1 x 105 labeled HL-60 cells in the assay buffer to each well and allow to adhere to the E-selectin for 30 to 45 minutes at 37 ° C. The plates are then gently washed 3 to 4 times with the assay regulator to remove unbound cells. Adherent cells are quantified by fluorescence measurement (Cytofluor 2350 system). Fluorescent Labeling of HL-60 Cells with BCECF-AM: HL-60 cells are cultured in Iscove medium supplemented with 20 percent fetal calf serum, glutamine, and non-essential amino acids. One day before the experiment is performed, the cells are subcultured (lxlO6 cells / milliliter). Cells (1 x 106 cells / milliliter) are labeled by incubation with 5 micrograms / milliliter of BCECF-AM (diluted from the supply in dimethyl sulfoxide) for 20 minutes at 37 ° C in phosphate-buffered serum.

Materials: ELISA plate: Nunc. Im uno Píate MaxiSorp (439454) HL-60 cells: obtained in the ATTC E-selectin catalog purified by affinity: each batch of E-selectin is functionally tested to determine the appropriate concentration to be used in the assay. Fluorescent dye: bis-carboxyethyl-carboxyfluorescein acetoxymethyl ester (BCECF-AM) available from Molecular Probes.

In this essay, the compounds of formula I inhibit the adhesion of HL-60 cells to E-selectin, in a concentration of 0.3 to 10 mM. The compounds of Examples 12, 13, and 18 inhibit the adhesion of HL-60 cells to E-selectin at a rate of 100 percent and 80 percent (for the latter two) at a concentration of 10 mM. Cell-free assay of sLe'-pol? Jaero / E-selectin. The rat CK antibody against mouse (10 micrograms / milliliter in carbonate / bicarbonate buffer, pH 9.5), prepared from the rat hybridoma cell line 187.1 (ATCC), is coated on microtiter plates overnight 4 ° C. The plates are washed twice with the assay regulator (20 mM HEPES, pH 7.4, containing 150 mM NaCl and 1 mM CaCl 2), blocked for 8 hours at 4 ° C with 3 percent bovine serum albumin in the regulator of the test, and washed three times. E-selectin mouse CK fusion protein (3 micrograms / milliliter in assay buffer) is added to each well, followed by overnight incubation at 4 ° C. A complex is formed between the biotinylated sLea-polymer (polyacrylic amide type glycoconjugate, Syntesome GmbH, Munich, Germany) containing 20 mole percent sLea (= 0.81 micromoles sLea / milligram polymer), and streptavidin peroxidase (Boehringer Mannheim, Germany), by incubation 20 microliters of sLea-polymer (1 milligram / milliliter) with 80 microliters of streptavidin peroxidase, 20 microliters of fetal calf serum, and 80 microliters of the assay regulator without CaCl2 for two hours at 37 ° C (the complex of sLea- Streptavidin polymer / peroxidase previously formed is stable for several months at 4 ° C). Then the complex is diluted to 3: 10,000 in the assay regulator, and added to the E-selectin coated wells together with the compound to be tested. The complex is allowed to sit for 2 hours at 37 ° C before the plates are washed twice with cold assay regulator. A solution of ABTS peroxidase substrate (Biorad) is added to the cavities, and the reaction is stopped after 10 minutes. The bound sLea-polymer complex is determined by measuring the optical density at 405 nanometers in a microplate reader. In this test, the compounds of formula I inhibit the sLea-polymer / E-selectin binding interaction when used in a concentration of 0.07 to 10 nM. The compounds of Examples 12, 13, and 18 inhibit the SLea-polymer / E-selectin binding interaction at an ICr.o (50 percent inhibition) of 1 mM, 10 mM, and 0.5 mM, respectively. c) Live tests. A compound of the formula I can be used as a one-for-one replacement for live treatments of SLex, for example, as described in the rat / cobra venom model by Mulligan et al., Nature, 36: 149-151 (1993) or feline model of myocardial ischemia / reperfusion injury by Murohara et al., Cardiovascular Research 30, 965-974 (1995). The compounds of the formula I show a beneficial effect in these models.

Accordingly, the compounds of the formula I are useful in the treatment and / or prevention of disorders or diseases that are mediated by the binding of selectins in cell adhesion, particularly E-selectin, for example, acute inflammatory or autoimmune diseases. or chronicles such as rheumatoid arthritis, asthma, allergy conditions, psoriasis, contact dermatitis, adult respiratory distress syndrome, inflammatory bowel disease, and ophthalmic inflammatory diseases, infection diseases such as septic shock, traumatic concussion, thrombosis, and conditions of inappropriate platelet accumulation, cardiovascular diseases such as heart attacks, reperfusion injury, multiple sclerosis, and neoplastic diseases including conditions of metastasis, embolisms, and acute or chronic rejection of organ or tissue transplants. For the above uses, of course, the required dosage will vary depending on the form of administration, the particular condition to be treated, and the desired effect. However, in general satisfactory results will be achieved at dosing concentrations of approximately 0.5 to 80 milligrams / kilogram of body weight of the animal. The daily dosage concentrations suitable for larger mammals, for example humans, are of the order of about 100 milligrams to 1.5 grams / day, conveniently administered once, in dosages divided 2 to 4 times a day, or in a form of sustained release. The unit dosage forms suitably comprise from about 25 milligrams to 0.750 grams of a compound of formula I, together with a pharmaceutically acceptable diluent or carrier therefor. The compounds of the formula I can be administered in free form or in a pharmaceutically acceptable salt form. These salts can be prepared in a conventional manner, and exhibit the same order of activity as the free compounds. The present invention also provides a pharmaceutical composition comprising a compound of the invention, in free base form or in pharmaceutically acceptable salt form, in association with a pharmaceutically acceptable diluent or carrier. These compositions can be formulated in a conventional manner. The compounds of the invention can be administered by any conventional route, for example, parenterally, for example in the form of injectable solutions or suspensions, or in a nasal or suppository form. In accordance with the foregoing, the present invention further provides: a) a compound of the invention or a pharmaceutically acceptable salt for use as a pharmaceutical product; b) a method for preventing or treating the disorders indicated above, in a subject in need of such treatment, said method comprising administering to this subject an effective amount of a compound of the formula I, or a pharmaceutically acceptable salt; c) a compound of the invention or a pharmaceutically acceptable salt for use in the preparation of a pharmaceutical composition for use in the method as in b) above.

Claims

1. A compound of the formula I where: i) R is CH3, and whether Ri is a radical of the formulas (ai) or (a2) (to.) where: m is 2 or 3; n is 2 or 3; M is a cation; R2 is H or a saturated or unsaturated hydrocarbon residue with up to 20 carbon atoms, which is optionally in the? a formyl group, or an acetal group of alcohol of 1 to 4 carbon atoms or diol acetal of 2 to 4 carbon atoms; R3 is H, -CH2OH, or -CH2CH2OH; and R 4 is H, alkyl of 1 to 4 carbon atoms, -CH2OH, -CH2CH2OH, or -CH2CH2CH2OH, with the proviso that: 1) one of R3 and R4 is H, and 2) when R4 is H, R3 is -CH2OH or -CH2CH2OH, and 3) when R3 is H, R 4 is CH 3, -CH 2 OH, -CH 2 CH 2 OH or -CH 2 CH 2 CH 2 OH; or else: Ri is a radical of formula (b): OR where: R5 is: OOC-CH. CH-CO-NH-I ^ MOOC- _ alkyl (b3) (b2) MOOC - where: p is 1 or 2; q is 2 or 3; r is 1 or 2; Re is H, NH 2, or -NHR .., wherein R x is an amino protecting group; R7a is -CH2OH, -CH2CH2OH, or -CH (OH) -CH2OH, and R7b is H, or each of R7a and Rib is CH2OH; Rn is H, or -OH; Rp is - (CH2) j-COOM or -S03M, where j is 1, 2, or 3; and M is as defined above; the second hydroxy substituent of the phenyl group being in (b4) at any meta position; or R? is a radical of formula (c): O-CH-CH, I (C) R, - CO-NH-CH-CORβ wherein: R8 is OM-L, OR1, Rs-Rp, or -NHRy, wherein Mx is a cation, R14 is a saturated or unsaturated hydrocarbon residue, Ra is a spacer group, Rp is a phosphatidyl residue, and Ry is a lipophilic residue; Y MOOC - C (c,) (c2) 0-C, -4 alkyl (c,) (C where: s is 1 or 2, t is 1 or 2, v is 2 or 3, M, R6, Rn, and R13 are as defined above; and R10a is "CH2OH, -CH2CH2OH, OR -CH (OH) -CH2OH, and R10b is H, or each of R10a and R10b is CH2OH, the second hydroxy substituent being of the phenyl group at (c2) at any meta position or where: (ii) R is OH, and R is a radical of formula (d): 2a r I H I "(d) MOOC-CH, -) - CH-CO-N-C-CO-NH-CH 2-I '12b wherein: w is 1 or 2; R12a is -CH (0H) - (CH2) x-0H, and R12b is H, or each of R12a and? 2b is independently -CH20H or -CH2CH20H; x is 2 or 3; and R6 and M are as defined above.

2. A compound of the formula I according to claim 1, wherein R is CH3, and Rj. is a radical of the formula (c), where R9 is a radical (c ^ or (c3)

3. A compound of the formula I according to claim 1, wherein Rj is OH, and R is a radical of the formula (d)

4. A compound of the formula: where M is a cation.

5. A compound of the formula: OH OH where M is a cation.

6. A compound of the formula: where M is a cation.

7. A process for the production of a compound of the formula I according to claim 1, which comprises removing at least one protecting group that is present in a compound of the formula I in protected form, and when required, recovering the compounds of formula I thus obtained, in free form or in salt form.

8. A compound according to any of claims 1 to 6, for use as a pharmaceutical product.

9. A pharmaceutical composition comprising a compound according to any of claims 1 to 6, together with a pharmaceutically acceptable diluent or carrier therefor.

10. A method for preventing or treating disorders or diseases that are mediated by the binding of selectins in cell adhesion, in a subject in need of such treatment, said method comprising administering to this subject an effective amount of a compound in accordance with any of Claims 1 to 6.