BACKGROUND OF THE INVENTION Cell adhesion is a process by which cells associate with each other, migrate towards a specific target or localize within the extra-cellular matrix. These interactions are mediated by specialized molecules called cell adhesion molecules (CAM). CAMs have been demonstrated to participate in various cell-cell, cell-extracellular matrix, and platelet interactions. They influence the adhesion of leukocytes to the vascular endothelium, their transendothelial migration, retention at extravascular sites and activation of T cells and eosinophils. These processes are central to the pathogenesis of inflammatory and autoimmune diseases. Therefore, adhesion molecules are considered as potential targets to treat such disorders.

CAMs can be classified into three groups-integrins, selectins and the immunoglobulin superfamily. Out of these, integrins are key mediators in the adhesive interactions between hemopoietic cells and their microenvironment. They comprise of alpha- beta heterodimers and integrate signals from outside of the cells to inside and vice versa.

Integrins can be classified on the basis of the alpha and beta subunits they contain. For example, beta-1 subfamily contains beta-1 subunit noncovalently linked to one of the 10 different alpha subunits.

The alpha-4 beta-1 integrin, also known as VLA4 (very late activation antigen 4), is a member of beta 1 integrin family and consists of alpha-4 and beta-1 subunits. VLA4 interacts with two specific ligands-the vascular cell adhesion molecule (VCAM-1) and the the CS1

region of fibronectin. Adhesion mediated by VLA4 is central to the process of transendothelial migration of leukocytes. Ligation of VLA4 is followed by gross rearrangement of the cytoskeleton leading to flattening of cells along the blood vessel wall followed by expression of specific molecules which digest the endothelial cell wall and diapedesis. Once in the extraluminal region, the interactions of VLA4 with extracellular fibronectin play a crucial role in migration to the site of inflammation, T cell proliferation, expression of cytokines and inflammatory mediators. In addition, VLA4 ligation provides costimulatory signal to the leukocytes resulting in enhanced immunoreactivity. Therefore, it is expected that VLA4 antagonists would ameliorate the immune response through twofold actions-inhibition of T cell recruitment at the site of inflammation and inhibition of costimulatory activation of immune cells.

In this respect, inhibitors of VLA4 interactions have demonstrated beneficial therapeutic effects in several animal models of inflammatory, and allergic diseases including sheep allergic asthma (Abraham et al, J. Clira. Invest., 93,776 (1994) ), arthritis (Wahl et al, J.

Clin. Invest. 94 655 (1994) ) ; experimental allergic encephomyelitis (Yednock et al, Nature (load), 356, 63 (1992) and Baron et al, J. Exp. Med., 177, 57 (1993) ) ; contact hypersensitivity (Chisolm et al, Eur J. Immunol., 23,682 (1993) ); type I diabetes (Yang et al, Proc. Natl. Acad. Sci. (USA), 90,10494 (1993) ) and inflammatory bowel disease (Podolsky et al, J. Clin. Invest., 92,372) (1993).

Region of CS 1 moiety of fibronectin involved in the interaction with VLA4 was identified as the tripeptide Leu-Asp-Val, also known as LDV (Komoriya et al, J. Biol. Chem.

266, 15075 (1991) ). Taking a lead from this, several peptides containing the LDV sequence were synthesised which have shown to inhibit the in vivo interaction of VLA4 to its ligands.

(Ferguson et al, Proc. Natl. Acad. Sci., (USA), 88, 8072 (1991); Wahl et al, J. Clin. Invest., 94,655 (1994); Nowlin et al, J. Biol. Chem. l 268 (27 !, 20352 (1993) and PCT publication W091/4862).

Despite these advances, there remains a need for small and specific inhibitors of VLA4 dependent cell adhesion molecules. Ideally such inhibitors should be water soluble

with oral efficacy. Such compounds would provide useful agents for treatment, prevention or suppression of various inflammatory pathologies mediated by VLA4 binding.

Isopropylidene and benzylidene groups are commonly used protective groups in carbohydrate chemistry. Although both these groups are introduced into a molecule under similar conditions, the location of the protection can be quite different. The reason for this difference is directly related to the stability of each protected molecule. Since protection normally occurs under conditions which allow reversibility, reaction proceeds until equilibrium is reached. The distribution of products at equilibrium is determined by their relative thermodynamic stabilities. In other words, these reactions are thermodynamically controlled. Benzylidene groups tend to be part of six-membered ring acetals, while the ketals resulting from acetonation generally are 5-membered rings. The difference is attributed to the effect of the methyl and phenyl substituents on the stability of the particular ring systems.

The prior art reveals that in the case of D-glucose, which is blocked in its furanose ring structure, the 1, 2- and 5,6-hydroxyl groups can be blocked using with isopropylidene or cyclohexylidene blocking group with the 3-position left open to undergo derivatization.

Therapeutic activity of hexoses and their derivatives are also disclosed in the prior art.

2, 3-O-isopropylidene-1-O-substituted-5, 6-dideoxy-5-N- [4- (2-hydroxy-2- oxoethyl) phenylamino carbonyl] amino-0, L-gulofuranosides (also known as 2, 3-O- isopropylidene-1-O-substituted-5, 6-dideoxy-5-N-{[4-(2-hydroxy-2-oxoethyl) phenyl] amino carbonylamino}-c2, D-epimannofuranoside are among compounds described in WO 00/42053, and are being extensively investigated clinically as cell adhesion inhibitors and for the prevention of cell adhesion mediated pathologies including inflammatory and autoimmune diseases.

SUMMARY OF THE INVENTION The base addition salts described herein are useful, inter alia, for the inhibition and prevention of cell-adhesion mediated inflammatory and autoimmune disorders.

Thus, pharmaceutical compositions containing these compounds and the methods of treating cell adhesion mediated inflammatory and autoimmune disorders are also made possible.

Base addition salts of particular ß-L-gulofuranoside compounds (shown as Formula I), have demonstrated protective effect in experimental models of airway hyper reactivity and airway inflammation.

FORMULAI wherein, n is 1,2 or 4, An+ is Li+, Na+, K+, Mg2+, Ca2+ or Ti4+ and R is straight chain or branched Cl to Cls alkyl, alkene, alkyne. , aryl, substituted aryl or alkylaryl.

Standard assays have been performed on compounds described herein to ascertain their efficacy on airway reactivity, and airway inflammation. The efficacy of the disclosed compounds in the experimental models of airway reactivity and inflamation indicate potential benefit in the treatment of asthma and Chronic Obstructive Pulmonary Disease (COPD).

Thus it has been found that the base addition salts of the disclosed compounds (Formula I) have shown superior"Maximum Inhibitory Effect"than the corresponding free acid forms.

Further the salts of the free acid forms are easier to handle than the free acid forms in the manufacture of pharmaceutical dosage forms.

Particular salts of the compounds disclosed herein are those wherein Jazz is Na+, K+, Mg2+ and Ca2+. Other particular salts are Na+ and Mg2+. The present invention also includes within its scope process of making prodrugs of the compounds of Formula I. In general, such prodrugs will be functional derivatives of these compounds which readily get converted in vivo into the defined compounds. Conventional procedures for the selection and preparation of suitable prodrugs are known.

The invention also include processes of making enantiomers, diastereomers, N-oxides as well as metabolites of the disclosed compounds having the same type of activity. The processes can further be adopted to include processes of making pharmaceutical compositions comprising the molecules of Formula I or prodrugs, metabolite enantiomers, diastereomers, N-oxides, pharmaceutically acceptable solvates or polymorphs in combination with pharmaceutically acceptable carriers and optionally included excipients.

Other aspects of the invention will be set forth in the description which follows, and in part will be apparent from the description, or may be learned by the practice of the invention.

DETAILED DESCRIPTION OF THE INVENTION The salts of Formula I are prepared by reacting a compound of Formula II with a base capable of releasing a cation An+ as shown in Scheme I, wherein n is 1,2 or 4, An+ is Li+, Na+, K+, Mg2+, Ca2+ or Ti4+, and R is straight chain or branched Cl-ls alkyl, alkene, alkyne, aryl, substituted aryl, or alkylaryl to give a compound of the Formula I which is then isolated by known methods.

Scheme I NHwCH2COOH oF NH 0 """"OO OR FORMULAI Base 9 FNH<CH2COO 0= NH 0 oxo OR

FORMULAI The starting compound of Formula II is prepared by following processes as described in WO 00/42053.

Examples of bases capable of releasing the cation An+ and examples of reaction conditions are given below: (a) Compounds of Formula I, wherein Jazz is Li+, Na+ or K+ are prepared by treating the free acid of Formula II with LiOH, NaOH or KOH in an aqueous or nonaqueous medium; or

(b) by treating the free acid of Formula II with LiORz NaORI or KORI, wherein R1 is an alkyl group containing 1-4 carbon atoms, in a non aqueous medium, such as an alcohol.

(c) Compounds of Formula I (wherein Jazz is Mg2+, Ca2+ or Ti4+) are prepared by treating the free acid of Formula II with Mg (OR1) 2, Ca (ORz) 2 or Ti (ORI) 4 wherein R1 is an alkyl group containing 1-4 carbon atoms, in a non-aquous solvent such as an alcohol.

(d) A compound of Formula I may be converted to another salt of the same Formula by exchanging the cation. For example, Na+ as a counter ion may be exchanged for Ca2+ or Mg Illustrative examples of the group R1 are CH3, C2H5, n-C3H7, n-C4H9, i-C4H9, sec. - C4H9 and ter-C4H9.

The examples mentioned below demonstrate the general synthesis and provide specific preparations of particular compounds. The examples are given to illustrate the details of invention and do not limit the scope of the present invention.

EXAMPLE 1: Preparation of 2, 3-O-isopropvlidene-1-O-dodecyl-5, 6-dideoxy-5-Nf4- (2- methoxv-2-oxoethylphenyllamino carbonyl amino} D-5-epimannofuranoside Step 1: 2, 3-O-isopropylidene-l-O-dodecyl-6-deoxy-5-p-tosyl-of, D-mannofuranoside 2, 3-O-isopropylidene-l-O-dodecyl-6-deoxy-ct, D-mannofuranoside (prepared according to the method reported in U. S. Patent No. 5,360, 794) (6.0 gm) was dissolved in pyridine (5 ml) and cooled to 0-5 °C. To this, was addedp-toluenesulfonyl chloride (5.3 g) portionwise with stirring. After about 7 hours, the solvent was removed under vacuum to obtain the residue which was extracted with ethyl acetate, washed with saturated sodium bicarbonate solution and brine. The ethyl acetate layer was dried over anhydrous sodium sulphate and the solvent was evaporated in vacuo to obtain an oily product which was purified by column chromatography using ethylacetate-hexane (5: 95) as eluent to afford the title compound in 82% yield.

The compounds prepared analogously were as follows: 2, 3-O-isopropylidene-l-O-butyl-6-deoxy-5-p-tosyl-a, D-mannofuranoside 2, 3-0-isopropylidene-1-0-hexyl-6-deoxy-5-p-tosyl- α,D-mannofuranoside 2, 3-O-isopropylidene-1-O-heptyl-6-deoxy-5-p-tosyl- α,D-mannofuranoside 2, 3-O-isopropylidene-1-O-decyl-6-deoxy-5-p-tosyl- α,D-mannofuranoside Step 2: 2, 3-O-isopropylidene-1-O-dodecyl-5, 6-dideoxy-5-azido-a, D-5- epimannofuranoside A mixture of the compound obtained from step 1 (9.0 g), sodium azide (9.0 g), and DMF (50 ml) was heated at 100 OC for about 48 hours. The solvent was removed under vacuum and the residue obtained was dissolved in ethylacetate. It was washed with water (2 x 50 ml) the organic layer was dried over anhydrous sodium sulphate and the solvent was removed under vacuum. The crude material was purified by column chromatography by eluting with a mixture of 2% ethyl acetate in hexane to get an oily product in 42% yield.

The compounds prepared analogously were as follows: 2, 3-O-isopropylidene-1-O-butyl-5, 6-dideoxy-5-azido-o, D-5-epimannoffiranoside 2, 3-O-isopropylidene-1-O-hexyl-5, 6-dideoxy-5-azido- α,D-5-epimannofuranoside 2, 3-O-isopropylidene-1-O-heptyl-5, 6-dideoxy-5-azido-ot, D-5-epimannofuranoside 2, 3-0-isopropylidene-1-0-decyl-5, 6-dideoxy-5-azido-c2, D-5-epimarmofuranoside Step 3: 2, 3-O-isopropylidene-1-O-dodecyl-5, 6-dideoxy-5-amino-a, D-5-epimanno furanoside To a suspension of lithium aluminium hydride (LAH) (2.0 gm) in dry THF (50 mL) at 0-5 C, was added a solution of the above compound obtained from step 3 (3.0 gm in 10 mL of THF) dropwise. Once the addition was over, the reaction mixture was further stirred at room temperature for about 2 hours. Excess LAH was decomposed by addition of an ice- water mixture. The reaction mixture was then filtered through celite, washed with THF and the solvent was evaporated in vacuo. The residue was dissolved in ethyl acetate, washed with water and brine. The solvent was dried over anhydrous sodium sulphate and the solvent was

removed under reduced pressure. The crude product was purified by column chromatography using ethylacetate-hexane (50: 50) as an eluent to yield pure product in 61% yield.

The compounds prepared analogously were as follows: 2, 3-O-isopropylidene-1-O-butyl-5, 6-dideoxy-5-amino-o, D-5-epimannofuranoside 2, 3-0-isopropylidene-1-0-hexyl-5, 6-dideoxy-5-amino- cD-5-epimannofuranoside 2, 3-O-isopropylidene-1-O-heptyl-5, 6-dideoxy-5-amino-ot, D-5-epimannofuranoside 2, 3-O-isopropylidene-1-O-decyl-5, 6-dideoxy-5-amino- α,D-5-epimannofuranoside Step 4: 2, 3-O-isopropylidene-1-O-dodecyl-5, 6-dideoxy-5-N-{[4-(2-methoxy-2-oXo ethyl) phenyl] amino carbonyl amino}-ot, D-5-epimannofuranoside To a cold (0-5°C) solution of the amine obtained from step 3 (0.5 g) in dry methylene chloride, was added the methyl ester of isocyanate-4-phenyl acetic acid (250 mg) dissolved in dry methylene chloride at 0-5°C and the reaction was stirred at same temperature for about 3 hours. Excess of methylene chloride was added to it, washed with water and brine.

The methylene chloride layer was dried over anhydrous sodium sulphate and the solvent was removed under vacuum to get an oily crude product. This product was purified by column chromatography over silica (230-400 mesh) and eluted with ethyl acetate-hexane (20: 80) mixture to get a pure white solid in 80% yield.

The compounds prepared analogously were as follows : 2, 3-O-isopropylidene-1-O-butyl-5, 6-dideoxy-5-N- { [4- (2-methoxy-2- oxoethyl) phenyl] aminocarbonyl amino}-a, D-5-epimannofuranoside 2, 3-O-isopropylidene-1-O-hexyl-5, 6-dideoxy-5-N-{[4-(2-methoxy-2- oxoethyl) phenyl] aminocarbonyl amino}-a, D-5-epimannofuranoside 2, 3-O-isopropylidene-1-O-heptyl-5, 6-dideoxy-5-N-{[4-(2-methoxy-2- oxoethyl) phenyl] aminocarbonyl amino}-a, D-5-epimannofuranoside 2, 3-O-isopropylidene-1-O-decyl-5, 6-dideoxy-5-N-{[4-(2-methoxy-2- oxoethyl) phenyl] aminocarbonyl amino}-α,D-5-epimannofuranoside

Example 2 : Preparation of 2, 3, 0-isopropylidene-1-0-dodecyl-5, 6-dideoxy-5-N-jr4- (2- hydroxy-2-oxoethyl ! phenyllamino carbonyl amino} D-5-epimannofuranoside A mixture of the ester (0.3 g) obtained in step 4 of example 1 and aqueous sodium hydroxide (1N, 10 ml) was heated at 50°C for about two hours. The reaction mixture was cooled to 0-5°C, acidified to pH3 with 3N HC1, a white solid separated out was filtered, which became an oil at room temperature. This anhydrous sodium sulphate and the solvent was removed in vacuum to get a crude oily product. The crude product was purified by flash column chromatography by eluting with ethyl acetate hexane mixture (35: 65) to get a low melting solid in 75% yield.

Example 3: Preparation of sodium salt of dodecyl 23-O-isopropylidene-56-dideoxy-5-N- [4- (2-hydroxy-2-oxoethyl) phenylaminocarbonyl] amino-a-L-gulofuranoside.

To a solution of dodecyl 2, 3,-O-isopropylidene-5, 6-dideoxy-5-N- [4- (2-hydroxy-2- oxoethyl) phenylaminocarbonyl] ß-L-gulofuranoside (54.8 g, 0.1 mole) in ethylalcohol (274 ml) was added sodium ethoxide (6.8 g, 0.1 mole, 1 equivalent) at room temperature and stirred for 1 hour to get a clear solution. Ethyl alcohol was removed under reduced pressure on rotary evaporator at 40-45°C. A foaming solid was obtained which was dried at 40-45°C under high vacuum for 8-10 hours. The foaming hygroscopic solid thus obtained was powdered under controlled humidity as an off-white hygroscopic solid, yield 57g, sodium content = 3.8-4. 2% w/w (Purity by HPLC=99.64%).

'HNMR (300 MHz), DMSO d6) # : 0.83-0. 87 (3H, t), 1.10-1. 12 (3H, d), 1.23-1. 52 (21H, bs), 1.38 (3H, s), 1.46-1. 48 (2H, m), 3.17 (2H, s), 3.29-3. 35 (1H, s), 3.54-3. 60 (1H, m), 3.76-3. 80 (1H, t), 3.80-4. 0 (1H, m), 4.49-4. 51 (1H, d), 4.70-4. 72 (1H, m), 4.89 (1H, s), 6.73-6. 75 (1H, d), 7.02-7. 05 (2H, d), 7.23-7. 26 (2H, d), 8.90 (1H, s) Mass: m/z = 571. 2 (MH, 549.3 (MH+ of acid) IR (DCM Film) Crri I :

Example 4: Preparation of potassium salt of dodecyl 2, 3-O-isopropylidene-5, 6-dideoxy-5-N- r4- (2-hydry-2-oxoethyl) phenylaminocarbonvllamino-a-L-gulofuranoside.

To a solution of dodecyl 2, 3-O-isopropylidene-5, 6-dideoxy-5-N- [4- (2-hydroxy-2- oxoethyl) phenylaminocarbonyl] amino-ßL-gulofuranoside (16.44g, 0. 03 moles) in methanol (85 ml) was added potassium hydroxide (1 mol. eq. ) at room temperature and stirred for 1 hour to get a clear solution. Methanol was removed under reduced pressure at 40-45°C on rotary evaporator. The foaming solid thus obtained was dried at 40-45°C under high vacuum for 8-10 hours. It was then powdered under controlled humidity to afford off white hygroscopic powder (yield 17g, potassium content = 6.3-6. 9% w/w).

The IHNMR spectrum (300 MHz), in DMSO d6 showed the following peaks: 6 : 0. 83- 0.85 (3H, t), 1.08-1. 11 (3H, d), 1.23-1. 25 (21H, bs), 1.38 (3H, s), 1.46-1. 47 (2H, m), 3.07 (2H, s), 3.33-3. 62 (2H, m), 3.77-3. 82 (1H, m), 3.9-4. 0 (1H, m), 4.48-4. 50 (1H, d), 4.69-4. 72 (1H, m), 4.87 (1H, s), 6.99-7. 01 (2H, d), 7.21-7. 24 (3H, m), 9.30 (1H, s) The mass spectrum showed: m/z = 549.3 (MH+ of acid) The IR spectrum (DCM film) showed the following signals: 3361.8 cm'\ 2925. 1 cm'\ 2854. 5 cm~l, 1657.3 cm' Example 5: Preparation of calcium salt of dodecyl 2, 3-O-isopropylidene-5 6-dideoxy-5-N- [4- (2-hydroxy-2-oxoethylphenylaminocarbonyl] amino-a-L-gulofuranoside.

To a solution of compound prepared in example 3 (l. lg, 1.93 mmol) in water (11 ml) calcium chloride (0.214, 1.93 mmol, 1 mol. Eqv. ) was added and stirred at room temperature for about lhour. Calcium salt was separated out as white solid. It was filtered, washed with water and dried under vacuum at 40-50°C (yield lg, calcium content = 3.52% w/w).

The IHNMR spectrum (300 MHz), in DMSO d6 showed the following peaks: 8 : 0.83-0. 87 (3H, t), 1.10-1. 20 (3H, d), 1.21-1. 25 (21H, bs), 1.37 (3H, s), 1. 48 (2H, bs), 3.22 (2H, s), 3.34-3. 59 (2H, m), 3.74-3. 78 (1H, m), 3.94-4. 02 (1H, m), 4.49-4. 51 (1H, d), 4.69- 4.72 (1H, m), 4.89 (1H, s), 6.52-6. 55 (1H, d), 7.05-7. 08 (2H, d), 7.24-7. 27 (2H, d), 8.74 (1H, s) The Mass spectrum showed: m/z = 549.2 (MH+ of acid) The IR spectrum (film, DCM) showed the following signals: 3344.7 cm'\ 2926. 8 cm-1, 2855 cm'\ 1652. 3 cm' Example 6: Preparation of magnesium salt of dodecyl 2, 3-O-isopropylidene-5, 6-dideoxy-5- N-[4-(2-hydroxy-2-oxoethyl)phenylaminocarbonyl]amino-ß-L-gu lofuranoside.

Magnesium turnings (0.328g, 0.014 mole, 0.5 mol. Eqv. ) were added to 60 ml of methyl alcohol and it was heated at 50-60°C for about 1 hour. A hazy solution thus obtained was added to a solution of dodecyl 2, 3-O-isopropylidene-5, 6-dideoxy-5-N- [4- (2-hydroxy-2- oxoethyl) phenylaminocarbonyl] amino-ß-L-gulofuranoside in methanol (15g, 0.027 moles in 60 ml of methanol). The reaction mixture so obtained was stirred for about 2 hours at 50- 60°C. It was then filtered and methanol was removed from the filtrate on rotary evaporator at 50-55°C to afford off-white solid, which was dried under vacuum for 2 hours (Yield 15 g, Mg content = 2.04-2. 26% w/w).

The 1HNMR spect-um in DMSO d6, (300 MHz) showed the following peaks 6 : 0.83- 0.87 (3H, t), 1.11-1. 14 (3H, d), 1.23-1. 25 (21H, bs), 1.37 (3H, s), 1.46-1. 48 (2H, m), 3.17 (2H, s), 3.35-3. 41 (2H, m), 3.54-3. 58 (1H, m), 3.76-3. 77 (1H, m), 3.94-4. 0 (1H, m), 4.50- 4.52 (1H, d), 4.70-4. 72 (1H, m), 4.90 (1H, s) 6.15 (1H, bs), 7.05-7. 08 (2H, d), 7.23-7. 26 (2H, d), 8.40 (1H, s) The mass spectrum showed: m/z = 549.2 (MH+ of acid)

PHARMACOLOGICAL ACTIVITY The compounds of the present invention have demonstrated protective effect in experimental models of airway hyperreactivity and airway inflammation. Standard assays have been performed on compounds of the present invention to ascertain their efficacy on airway reactivity, and airway inflammation. These include: 1. Ovalbumin-induced airway reactivity and airway inflammation in guinea pig.

2. LPS induced airway reactivity and airway inflammation in rat.

Enhanced airway reactivity and airway inflammation are common features of respiratory diseases like bronchial asthma and chronic obstructive pulmonary disease (COPD) (O'Byrne and Postma; 1999; Rutgers et al., 2001). Guinea pigs sensitised and challenged with ovalbumin exhibit intense airway reactivity to histamine with accompanying influx of eosinophil into the airways, a model representative of bronchial asthma (O'Byrne and Postma; 1999). By contrast, following LPS exposure rats develop airway reactivity with accompanying neutrophil influx into the airways, a condition simulating COPD (Wheeldon et al. , 1992). Thus, the efficacy of compounds in above-described models of airway reactivity and inflammation may indicate potential benefit in the treatment of asthma and COPD.

Guinea pig airway reactivity and inflammation 1. Sensitization of guinea pigs Guinea pigs were sensitised on days 0,7 and 14 with 50 ug ovalbumin and 10 mg aluminium hydroxide injected intraperitoneally. On days 19 and 20 guinea pigs were exposed to 0. 1% w/v ovalbumin or PBS for 10 min. On day 21 guinea pigs were exposed to 1% ovalbumin for 30 min. One group of guinea pigs were exposed to intraperitoneal ovalbumin as described, but were challenged with phosphate buffered saline (PBS) and served as control. Mepyramine 5 mg/kg, i. p. , was administered to each guinea pig 30 min before ovalbumin/PBS challenge.

2. Administration of compounds Treatment with any of the following TRIS salts of compound of Example 2, Example 3 montelukast, dexamethasone or vehicle once daily was started from day 18 onwards and continued for 5 days (i. e. on days 18, 19,20, 21,22). Ovalbumin/PBS challenges were performed 2 hours after different drug treatment.

3. Airway Reactivity Measurement: Histamine dose response On day 22,2 hours after drug treatment or vehicle administration, animals were transferred to a whole body plethysmograph (Buxco Electronics, USA) to study the airway reactivity as described in Hamelmann et al. , (1997) and Chong et al. (1998). Animals were allowed to acclimatise in the body box and the basal PenH value (an index of airway resistance) was recorded using Bettsystem XA software, (Buxco Electronics, USA). This was followed by successive challenges, each of 2 min duration, with PBS (vehicle for histamine) or histamine (0.1, 0.3, 0.6, 1,1. 5,3, 6 and 10 mg/ml). Recording was stopped at once airway resistance increased 3 fold over PBS response.

4. Bronchoalveolar lavage (BAL) After histamine dose response study, animals were sacrificed using thiopentone sodium (150 mg/kg/i. p. ). Tracheas were cannulated and BAL was performed using Hank's Buffer salt solution (HBSS) (5 ml x 10 times). Lavage fractions were pooled and centrifuged at 3000 rpm for 5 min, at 4°C. The pellets were collected and resuspended in 1-ml HBSS.

Total leukocyte counts were performed in the resuspended sample using a hemocytometer. A smear was prepared using the resuspended BAL fluid on a glass slide and was left to dry.

Dried slides were kept on a staining tray and flooded with Leishmann's stain for 1 min. The stain was then diluted in the ratio of 1: 2 with buffer (pH 7) and left for 10 min. Slides were washed with tap water and left to dry and then differential leukocyte counts were performed.

5. Data analysis : i. Airway Reactivity PC_ computation PenH values in each guinea pig were obtained in the presence of PBS and different doses of histamine. PenH, at any chosen dose of histamine was, expressed as percent of PBS

response. The PenH values thus calculated and the corresponding histamine dose were fed into Graph Pad Prism software (Graphpad Software Inc. , USA) and using a nonlinear regression analysis PC200 (dose of histamine increasing PenH 3 folds of PBS value) values were computed.

ED computation PC200 values obtained in the presence of different doses of test compounds were expressed as percent protection using the formula given below. EDso value was obtained by linear regression analysis of concentration and percent protection data.

PC200TEST-PC200 OVA % Protection =-------------X 100 PC200 M AXPC200 OVA Where, PC200TEST= PC200 value obtained in the presence of a given dose of test compound PC200 M = Maximum PC200 value obtained in the presence of test compound PC200 OVA = PC200 value obtained in the absence of test compound ii. Airway Inflammation Eosinophil count in bronchoalveolar lavage fluid was expressed as percent of total leukocyte count. Inhibition of eosinophil influx into airway was computed using the following formula: EOSpVA-EOSTEST % Inhibition = ---------------------- X 100 EosovA-EoscoN Where, EosovA = Percentage of eosinophil in untreated ovalbumin challenged group EosTEST = Percentage of eosinophil in group treated with a given dose of test compound EOSCON = Percentage of eosinophil in group not challenged with ovalbumin EDso dose for inhibition of eosinophil influx was calculated by linear regression analysis of concentration and percent protection data. Table 1 shows the effect of Tris salt of compound of Example 2, Example 3 montelukast and dexamethasone on airway reactivity in guinea pig Compound Route of Airwa Reactivity administration PC-200 ED50 mg/k Compound of Example 3 Oral 1. 08 0. S5 TRIS salt of compound of Oral 1. 981 1. 59 Example 2 Montelukast Oral 1. 062 0. 04 Dexamethasone Oral 1. 49 1. 38

(1) Effect observed at 10 mg/kg ; (2) Effect observed at 0.1 mg/kg TRIS implies [tris (hydroxymethylaminomethane)] Table 2 shows the effect of Compound of Example 3, TRIS salt of compound of Example 2, montelukast and dexamethasone on airway inflammation in guinea pig Treatment Route of Airway Inflammation administration Inhibition of ED50 eosinophil influx (mg/kg) Compound of Example 3 Oral 77 % 1.07 TRIS salt of compound of Oral 59 %'1. 32 Example 2 Montelukast Oral 59 % 0. 054 Dexamethasone Oral 100 % 1. 78

(1) Effect observed at 10 mg/kg; Rat airway reactivity and inflammation 1. Administration of compounds Animals were dosed orally every day of five days with vehicle, Compound of Example 3, TRIS salt of compound of Example 2, montelukast and dexamethasone.

2. Sensitization of rats On the day of experiment, animals were exposed to LPS (100 ug/ml) for 40 min (Toward and Broadley, 2000). One group of vehicle treated rats were exposed to phosphate

buffered saline for 40 min. Two hours after LPS/PBS exposure, animals were placed inside a whole body plethysmograph and exposed to acetylcholine (1, 3,6, 12,24 and 48 mg/ml) aerosol for 2 min. Between each challenge a gap of 7 minutes was allowed. In order to minimise movement-related artifacts during the experiment, animals were sedated by intraperitoneal injection of diazepam (10 mg/kg, intraperitoneally) in 3% tween 80.

3. Bronchoalveolar lavage (BAD Twenty-four hours after acetylcholine dose response study, animals were sacrificed using thiopentone sodium (150 mg/kg/i. p. ). Tracheas were cannulated and BAL was performed using Hank's Buffer salt solution (HBSS) (5 ml x 10 times). Lavage fractions were pooled and centrifuged at 3000 rpm for 5 min, at 4°C. Pellet was collected and resuspended in 1-ml HBSS. Total leukocyte count was performed in the resuspended sample by using a hemocytometer. Smears were prepared using the resuspended BAL fluid on a glass slide and left to dry. Dried slides were kept on a staining tray and flooded with Leishmann's stain for 1 min. The stain was then diluted in the ratio of 1: 2 with buffer (pH 7) and left for 10 min. Slides were washed with tap water and left to dry and then differential leukocyte counts were done.

4. Data analysis : PenH values in each rat were obtained in the presence of PBS and different doses of acetylcholine. PenH, at any chosen dose of acetylcholine was, expressed as percent of PBS response. Total neutrophil were expressed as million cells/ml of BAL.

Protective effect on airway reactivity and inflammation were calculated using the following formula: ResponseLps-ResponseTEST % Inhibition =-----------------------------------X 100 ResponseLps-ResponsecoN Where, ResponseLps = PenH (% Initial) or airway inflammation (Total neutrophil count) in LPS challenged rats in the absence of drug treatment

ResponseTEST = PenH (% Initial) or airway inflammation (Total neutrophil count) in LPS challenged rats treated with a given dose of test compound ResponsecoN = PenH (% Initial) or airway inflammation (Total neutrophil count) in PBS challenged rats Table 3 shows the maximum inhibitory effects of TRIS salt of compound of Example 2, the compound of Example 3, montelukast, and dexamethasone on airway reactivity and airway inflammation in rat. Maximum Inhibitory Effect Treatment Route of Airway Airway Inflammation Administration Reactivity Compound of Example 3 Oral 86% 1 63% 2 TRIS salt of compound of Oral 100% 1 87% 2 Example 2 Montelukast Oral 38%1 58% Dexamethasone Oral 100%@ 38%

(1) Effect observed at 0.1 mg/kg dose; (2) Effect observed at 10 mg/kg dose; While the present invention has been described in terms of its specific embodiments, certain modifications and equivalents will be apparent to those skilled in the art and are intended to be included within the scope of the present invention.