Title of Invention	A NOVEL COMBINATORIAL LIBRARY OF 3- SUBSTITUTED AMINO-3-GLYCOSYLATED PROPANOATES USEFUL AS ANTIFUNGAL AND ANTIBACTERIAL AGENTS
Abstract	The present invention relates to a combinatorial library of 3-substituted amino-3-glycosylated propanoates useful as antifungal and antibacterial agents. Efficiency has been the key goal in modern organic synthesis of biomolecules in drug design and discovery. Combinatorial synthesis is a method to prepare a large number of structurally distinct molecules where many experiments may be carried out at the same time. It has been achieved both by solution and solid phase methods using either divide & couple or split techniques.

Title of Invention

A NOVEL COMBINATORIAL LIBRARY OF 3- SUBSTITUTED AMINO-3-GLYCOSYLATED PROPANOATES USEFUL AS ANTIFUNGAL AND ANTIBACTERIAL AGENTS

Abstract

The present invention relates to a combinatorial library of 3-substituted amino-3-glycosylated propanoates useful as antifungal and antibacterial agents. Efficiency has been the key goal in modern organic synthesis of biomolecules in drug design and discovery. Combinatorial synthesis is a method to prepare a large number of structurally distinct molecules where many experiments may be carried out at the same time. It has been achieved both by solution and solid phase methods using either divide & couple or split techniques.

Full Text	The present invention relates to a novel combinatorial library of 3-substituted amino-3-glycosylated propanoate derivatives. More particularly the present invention relates to a novel combinatorial library of 3-substituted amino-3-glycosylated propanoate derivatives useful as antifungal and antibacterial agents. Background of the invention: Efficiency has been the key goal in modern organic synthesis of biomolecules in drug design and discovery. Combinatorial synthesis is a method to prepare a large number of structurally distinct molecules where many experiments may be carried out at the same time. It has been achieved both by solution and solid phase methods using either divide & couple or split techniques (A, Netzi, J.M. Ostresh, R.A. Houghten Chem Rev 1997. 97, 449). The parallel synthesis of numerous single compounds is considered to be most desirable nowadays and can be accomplished by sequential coupling to solid phase linked substrate if suitable combination of protecting and anchoring groups are applied. In comparison to other scaffolds carbohydrate derived scaffolds constitute template, which not only offer more utilizable functional groups but also numerous, alternatively addressable stereogenic centers (T. Wunberg, C. Kallus, T. Opatz, S. Henko, W. Schimdt, H. Kunz Angew. Chem. Int. Ed. 1998, 37, 2503). Despite the availability of effective and short course chemotherapy and BCG vaccine the tube -rcular baccilus continues to claim more lives than any other single infectious agent in the world. A number of substances other than the antibiotics have been proposed heretofore for use in treating tubercular and fungal infections, but they have not been used extensively for a number of reasons. Among them the increased incidence of multidrug resistant tuberculosis, its deadly synergy with human immunodeficiency virus and susceptibility to fungal infections in tuberculosis and resistance to existing antifungals are the prominent ones. Many potentially effective antitubercular and antifungal agents are toxic and cannot be safely administered to patients and still some of them have undesirable side effects. As a result of the foregoing and other deficiencies, it is apparent that the development of new class of nontoxic mechanism based broad spectrum antimicrobial agents has not been available for treating the tubercular and fungal infections. It has been discovered that certain glycosylated amino acids are associated with a number of biological activities as these provide important biological signals to the cells in order to combat the adverse conditions (Varki A., Glycobiology (1993), 3, 97 & Drickamen, K. In molecular glycobiology Flukada, M., Hindsgaul. O., Eds IRL, Oxford 1994, Ch2, pp53-87). Further sugar derivatives are known to offer better pharmacokinetic parameter to the drugs and at the same time they are less toxic in comparision to other class of compounds [J.F.Fisher, Harrison, A.W., Bundy G.L., K.F. Willinkson, Rush B.D., Ruwart M.J., J. Med. Chem(1991, 34, 3410 & Negre E., Driaf K.P., Veraneuil B, Tetrahedron Lett (1993), 34, 1027]. Combinatorial synthesis of small molecules has already gained momentum in drug discovery and research. However, the combinatorial syntheses of glycoconjugate have been very slow because of difficulty in their synthesis and structure elucidation. Further, glycoconjugates are known to be superior ligands with higher affinity than a natural carbohydrate ligand [P. M. St. Hilaire and M. Meldal Angew. Chem. Int. Ed (2000, 39, 1162)]. Hence, these carbohydrate based libraries can be essential tools in the development of carbohydrate based therapeutics particularly the anti-infectives. The main objective of the present invention is to provide novel combinatorial library of 3-substituted amino-3-glycosylated propanoate derivatives useful as antifungal and antibacterial agents. Another objective of the present invention is to provide a process for preparation of novel combinatorial library of 3-substituted amino-3-glycosylated propanoate derivatives of the general formula 7 (Figure-1). Yet another objective of the present invention is to provide compounds useful in the treatment of fungal and mycobacterial infections. Accordingly the present invention relates to novel combinatorial library of 3-substituted amino-3-glycosylated propanoates of the general formula 7 (figure-1) H2NC(CH2)nCHN wherein R is the alkyl chain consisting of 1-4 carbon alkyl, aralkyl groups or H, R\| is alkyl group or H; R2 is 1-4 carbon branched or unbranched alkyl groups; R3 is branched or unbranched alkyl, heterocycloalkyl or cyclogroup or H; n ranges from 0 to 4, R4 is 1 to 4 carbon branched or unbranched alkyl or acyl group , cycloalkyl or heterocycloalkyl groups, aroyl group where aromatic ring is substituted or unsubstituted; which comprises (i) preparing activated sieber amide resin by treating the sieber amide resin with 20-90 % piperidine-dimethyl formamide or piperidine, (ii) coupling the activated amide resin with Fmoc-aminoacids of the general formula 1, wherein R3 is branched or unbranched alkyl, heterocycloalkyl or cycloalkyl group or H; n ranges from 0 to 4, in presence of conventional coupling agent at a temperature in the range of 0-100 °C for a period in the range of 1 to 12 hrs to form amide linkage between the resin and amino acid of the general formula 1 producing the resin bound compound of the general formula 2 wherin value of R3 and n is same as given for formula 1, (iii) deprotecting the Fmoc group from the above substrate 2, by treating with 20-90 % piperidine in dimethyl formamide at 0 to 50 °C, followed by washing with dimethyl formamide and successively with dichloromethane to get the free amino group resin bound substrate, (iv)reacting alkyl- [3-(l,2-O-isopropylidene-3-O-substituted (alkyl or aralkyl)-l,4-pentofuranos-4-yl]-propenoates of the general formula 3 (figure-1) wherein R is the alkyl chain consisting of 1-4 carbon alkyl , aralkyl groups or H, Ri is alkyl group or H; R2 is 1-4 carbon branched or unbranched alkyl groups; to the aminoacyl resin in an organic solvent in presence of an organic base at the temperature in the range of 10-100 °C for a period 1-48 hrs, washing the resin bound compound of formula 4 wherein R, Rl and R2 are same as given for formula 2 and 3 as in step (iii); (v) reacting the resin bound compound of the general formula 4 either with a compound selected from alkyl or arylhalides/ acyl halides/ aliphatic / aromatic/ heteroaromatic acids or their acid chlorides of the general formula 5, wherein R4 is 1-12 carbon branched or unbranched alkyl chain , substituted or unsubstituted aromatic/heteroaromatic rings , X is CH2CI, CI-bBr , COC1 , COBr or COOH groups, n may vary from 0 to 12;, in dimethyl formamide/ dimethyl sulphoxide in presence of bases selected from diazabicycloundecene, pyridine, dimethyl amino pyridine at temperature ranging from 0 to 150 °C for a period 1-56 hrs and washing the reaction mixture as in case of step (iii) to give the resin bound compound of the general formula 6 wherein R, Ri, R2, R3 and R4 as stated above finally cleaving the resin bound glycoconjugates to give the compounds of the general formula 7 using 2-90% trifluoro acetic acid in dichloromethane in the temperature range of 0 to 40 °C; evaporating the solvents followed by lyophilization using tertiary butanol and water. The present invention provide a novel combinatorial library of 3-substituted- amino-3-glycosylated propanoates useful as antifiingal and antibacterial agents which comprises the following steps; (a) swelling of solid phase (sieber amide resin) with solvents such as dicloromethane, dimethyl formamide in nitrogen atmosphere; (b) Deprotection of Fmoc-group of the sieber amide resin to corresponding free amino sieber amide using 30-90% piperidine-DMF (dimethyl formamide); purging the excess solvents and washing the excessive reagents; (c) loading of the Fmoc-NH-amino acids of the general formula 1, on to a solid phase such as sieber amide resin using HOBt (1-hydroxy benzotriazole), DIPEA (di-isopropyl ethyl amine), TBTU [2-(lH- benzotriazole-1-yl) 1,1,3,3-tetramethyl uroniumtetrafluoroborate], DIPC (diisopropyl carbodimide) in an organic solvent at a temperaure in the range of 0-100 °C for a period ranging from 1 to 12 hrs; purging the excess solvents and washing the excessive reagents to give the resin bound compound of formula 2; (d) deprotecting the Fmoc group in compound 2 with 30-90% piperidine-DMF followed by washing with isopropanol and dichloromethane; (d) reacting alkyl- [3-(l,2-O-isopropylidene-3-O-substituted (alkyl or aralkyl)-l,4-pentofuranos-4-yl]-propenoates (R.P.Tripathi et.al. Indian Patent application No 48/DEL/2001) of the general formula 3 (figure-1) to the Fmoc deprotected aminoacyl resin of the general formula 2 in an organic solvent in presence of DBU (diazabicycloundecene) or triethyl amine at the temperature in the range of 10-100 °C for a period 1-48 hrs, purging the excess solvents and washing the excessive reagents to give the resin bound compound of the general formula 4; and (e) Functionalising secondary amino group with alkanoic acids, alkyl halides, acyl chlorides, hetero aromatic acids; of general formula 5 and 6 in an organic solvent at a temperature in the range of 0° C to 150° C for a period in the range of 1 to 56 hr, in presence of bases such as diazabicycloundecene, pyridine, dimethyl amino pyridine; purging the excess solvents and washing the excessive reagents to get the resin bound compound of the general formula 7; and (f) by cleaving the resin bound glycoconjugates using 2-50 % trifluoro acetic acid in dichloromethane at the temperature in the range of 0-40 ° C; evaporating the solvents at temperature 0-100 °C followed by freeze drying of the products so obtained to give the combinatorial library of glycoconjugates of general formula 7 (figure-1). In an embodiment of the present invention the reaction vessels used may be selected from syringes, automatecd robotic synthesiser, Merrifield's vessel, three way sintered vessel. In an embodiment of the present invention the solid phase may be selected from super acid sensitive resins sieber amide or rink amide resins. In another embodiment of the present invention the solvents used for swelling the resins may be selected from dicloromethane, dimethyl formamide and tetrahydrofuran. In yet another embodiment of the present invention the inert atmosphere used may be selected from nitrogen or argon atmosphere. In a further embodiment of the present invention the reagents used for deblocking the Fmoc group from the resins as well as resin bound compounds may be selected from 30-100 % piperidine in dimethyl formamide, triisopropyl silane, triethyl amine. In yet another embodiment of the present invention the solvent used in the reaction of Fmoc amino glycosylated propanoic acids and activated sieber amide resin, amino glycosylated propanoyl resin and Fmoc amino acid, aromatic or heteroaromatic acids may be selected from dimethyl formamide, dichloromethane, tetrahydrofuran. In yet another embodiment of the present invention the solvent used in the reaction of olefinic ester derived from sugars and resin bound amino acyl derivatives may be selected from alcohols such as methanol, ethanol, propanol, dimethyl formamide, tetrhydrofiiran, dioxane, dichloromethane etc. In a further embodiment of the present invention the catalyst used in the reaction between olefinic ester derived from sugars and resin bound amino acyl derivatives may be selected from triethyl amine, p-dimethyl amino pyridine, diazabicycloundecene or phase transefer catalyst such as tetra butyl ammonium bromide, Tetramethyl ammonium hydroxide, tetrabenzyl ammonium bromide. In a further embodiment of the present invention the reagent used in cleaving the compounds loaded on to resins may be 2-50% trifluoroacetic acid in dichloromethane. Thus the process used for this invention comprises the following steps (i) Sieber amide resin was placed in 24 wells in the reaction block of robotic synthesizer and treated with 20% piperidine-DMF (1 ml) twice for 5 min and 25 min. After this the resin in each well was washed with DMF (3x2 min), iPrOH (1x2 min) and DMF (3 x 2min). This was followed by transfer of one of the Fmoc amino acids such as P-alanine, leucine, isoleucine, a-amino butyric acid, y-amino butyric acid, valine or nor valine or their derivatives to the desired well and subsequently treated with, TBTU, DIC and HOBt in DMF and the reaction was carried out for 24 hrs. After completion the resins were again washed with DMF, dichloromethane, isopropanol separately. (iii)The Fmoc protecting group was deblocked using 20 % piperidine-DMF and washed with DMF, dichloromethane and solvent purged off and then treated with the desired alkyl- [3-(l,2-O-isopropylidene-3-O-substituted (alkyl or aralkyl)-l,4-pentofuranos-4-yl]-propionates in DMF and ethanol (1:1) in presence of triethyl amine or diazabicycloundecene (DBU) and the reaction wells were stirred for 24 hrs and the reaction vessels washed with DMF (3x2 min), iPrOH (1x2 min) and DMF (3 x 2min). The completion of reaction was monitored by negative ninhydrin test. (iv)This was followed by removal of Fmoc group in each well with 20% piperidine-DMF and washing the reaction vessels with DMF (3x2 min), iPrOH (1x2 min) and DMF (3 x 2min), drying the reaction wells. (v)The resin bound compounds in each well was treated separately with different alkyl halides, araalkyl halides or acyl halides in pyridine, dimethyl sulphoxide, DMF, THF etc in presence of bases such a triethyl amine, diazabicycloundecene, pyridine at 0 to 100 ° C and the reaction wells stirred for 20 hrs. The solvent and unreacted material was purged off. The reaction wells were washed with DMF (3x2 min), iPrOH (1x2 min) and DMF (3 x 2min) Finally the glycoconjugates were cleaved from the resin using mixture of 2% TFA, 94% dichloromethane and 4% triisopropylsilane. Evaporating the solvents and dissolving the products, so obtained, in t-Butanol/ Water (4:1) and lyophilizing the final compounds after deprotection and cleavage. In our copending patent application no 1272/del/01, claimed and disclosed the process for the preparation of novel combinatorial library of 3-substituted amino-3-glycosylated propanoates. From this application we have divided out this application The representative compounds of the formula 7 prepared by the process of the present invention are as follows: Ethyl-3- (N- [N- (benzoyl), N- p-alanylamido-l-yl]-amino-3- [(1R, 2R, 3S) 1,2-0- isopropylidene-3-O-methyl-1,4-pentofuranos-4-yl])-propanoate Ethyl-3- [N- (4-Nitrobenzoyl), N- (3-alanylamido-l-yl] -amino-3- [(1R, 2R, 3S) 1,2-0- isopropylidene-3-O-methyl-l, 4-pentofuranos-4-yl])-propanoate Ethyl-3- [N- (tetradecanoyl), N- p-alanylamido-1-yl] -amino-3- [(1R, 2R, 3S) 1,2-0- isopropylidene-3-O-benzyl-1,4-pentofuranos-4-yl])-propanoate Ethyl-3- [N- (dodecanoyl), N- P-alanylamido-1-yl] -amino-3- [(1R, 2R, 3S) 1,2-0-Isopropylidene-3-O-benzyl-1,4-pentofuranos-4-yl])-propanoate Ethyl-3- [N- (hexadecanoyl), N- P-alanylamido-1-yl] -amino-3- [(1R, 2R, 3S) 1,2-0-isopropylidene-3-O-benzyl-1,4-pentofuranos-4-yl])-propanoate Ethyl-3- [N- (dodecanoyl), N- p-alanylamido-1-yl] -amino-3- [(1R, 2R, 3S) l,2-O-isopropylidene-3 -O-methyl-1,4-pentofuranos-4-yl])-propanoate Ethyl-3- [N- (hexadecanoyl), N- p-alanylamido-1-yl] -amino-3- [(1R, 2R, 3S) 1,2-0-isopropylidene-3-O-benzyl-l, 4-pentofiiranos-4-yl])-propanoate Ethyl-3- [N- (hexadecanoyl), N- P-alanylamido-1-yl] -amino-3- [(1R, 2R, 3S) 1,2-0-isopropylidene-3-O-methyl-1,4-pentofuranos-4-yl])-propanoate Ethyl-3- [N- (tetradecanoyl), N-y-butyralamido-1-yl] -amino-3- [(1R, 2R, 3S) 1,2-O- isopropylidene-3-O-methyl-1, 4-pentofiiranos-4-yl])-propanoate Ethyl-3- [N- (octanoyl), N- p-alanylamido-1-yl] -amino-3- [(1R, 2R, 3S) 1,2-0-sopropylidene-3-O-methyl-1,4-pentofuranos-4-yl])-propanoate Ethyl-3- [N- (dodecanoyl), N-(a-aminobutyralmido)-l-yl] -amino-3- [{1R, 2R, 3S) l,2-O-Isopropylidene-3-O-methyl-l,4-pentofiiranos-4-yl])-propanoate Ethyl-3- [N- (hexadecanoyl), N- p-alanylamido-1-yl] -amino-3- [(1R, 2R, 3S) 1,2-0-isopropylidene-3-O-benzyl-l, 4-pentofiiranos-4-yl])-propanoate Ethyl-3- [N- (dodecanoyl), N- P-alanylamido-1-yl] -amino-3- [(1R, 2R, 3S) 1,2-0-isopropylidene-3-O-benzyl-1,4-pentofuranos-4-yl])-propanoate Ethyl-3- [N- (hexadecanoyl), N- p-alanylamido-1-yl] -amino-3- [(1R, 2R, 3S) 1,2-0-isopropylidene-3-O-benzyl-1,4-pentofuranos-4-yl])-propanoate Ethyl-3- [N- (hexadecanoyl), N- (3-alanylamido-l-yl] -amino-3- [(1R, 2R, 35) 1,2-0- isopropylidene-3-O-methyl-1,4-pentofiiranos-4-yl])-propanoate Ethyl-3- [N- (tetradecyl), N- P-alanylamido-1-yl] -amino-3- [(1R, 2R, 35) l,2-O- isopropylidene-3-O-methyl-1,4-pentofuranos-4-yl])-propanoate Ethyl-3- [N- (dodecanoyl), N- p-alanylamido-1-yl] -amino-3- [(1R, 2R, 35) 1,2-0- isopropylidene-3-O-methyl-1,4-pentofuranos-4-yl])-propanoate Ethyl-3- [N- (benzoyl), N- P-alanylamido-1-yl] -amino-3- [(1R, 2R, 35) 1,2-0- isopropylidene-3-O-methyl-1,4-pentofuranos-4-yl])-propanoate Ethyl-3- [N- (benzoyl), N- p-alanylamido-1-yl] -amino-3- [(1R, 2R, 3S) 1,2-0- isopropylidene-3-O-benzyl-l,4-pentofuranos-4-yl])-propanoate, Ethyl-3- [N- (benzyl), N- p-alanylamido-1-yl] -amino-3- [(1R, 2R, 3S) 1,2-0- isopropylidene-3-O-methyl-1,4-pentofiiranos-4-yl])-propanoate Ethyl-3- [N- (4-cyanobenzyl), N- P-alanylamido-1-yl] -amino-3- [(1R, 2R, 35) 1,2-0- isopropylidene-3-O-methyl-l, 4-pentofuranos-4-yl])-propanoate Ethyl-3- [N- (dodecanoyl), N-(hexamido-6-yl)] -amino-3- [(1R, 2R, 35) 1,2-0- isopropylidene-3-O-methyl-l, 4-pentofuranos-4-yl])-propanoate Ethyl-3- [N- (octanoyl), N-(hexamido-6-yl)] -amino-3- [(1R, 2R, 35) 1,2-0- isopropylidene-3-O-methyl-1,4-pentofuranos-4-yl])-propanoate Ethyl-3- [N- (benzoyl), N-(phenylalanylamido-2-yl)] -amino-3- [(1R, 2R, 35) 1,2-0- isopropylidene-3-O-methyl-l, 4-pentofuranos-4-yl])-propanoate Ethyl-3- [N- (benzyl), N-(phenylalanylamido-2-yl)] -amino-3- [(1R, 2R, 35) l,2-O- isopropylidene-3-O-methyl-l, 4-pentofUranos-4-yl])-propanoate Ethyl-3- [N- (benzyl), N-(phenylalanylamido-2-yl)] -amino-3- [(1R, 2R, 3S) 1,2-0- isopropylidene-3-O-benzyl-l, 4-pentofiiranos-4-yl])-propanoate Ethyl-3- [N- (4-cyanobenzyl), N-(phenylalanylamido-2-yl)] -amino-3- [(1R, 2R, 3S) 1,2-O-isopropylidene-3-O-methyl-1, 4-pentofuranos-4-yl])-propanoate Ethyl-3- [N- (benzyl), N-(propanamido-2-yl)] -amino-3- [(1R, 2R, 3S) 1,2-0- isopropylidene-3-O-methyl-1,4-pentofiiranos-4-yl])-propanoate Ethyl-3- [N- (benzoyl), N-(propanamido-2-yl)] -amino-3- [(1R, 2R, 3S) 1,2-0- isopropylidene-3-O-methyl-1,4-pentofuranos-4-yl])-propanoate Ethyl-3- [N- (dodecanoyl), N-(propanamido-2-yl)] -amino-3- [(1R, 2R, 3S) 1,2-0- isopropylidene-3-O-methyl-l, 4-pentofuranos-4-yl])-propanoate Ethyl-3- [N- (tetrdecanoyl), N-(propanamido-2-yl)] -amino-3- [(1R, 2R, 3S) 1,2-0- isopropylidene-3-O-methyl-1,4-pentofuranos-4-yl])-propanoate Ethyl-3- [N- (pentaEthyl-3- [N- (tetrdecanoyl), N-(propanamido-2-yl)] -amino-3- [(1R, 2R, 3S) l,2-O-isopropylidene-3-O-methyl-l, 4-pentofuranos-4-yl])-propanoate decanoyl), N-(propanamido-2-yl)] -amino-3- [{1R, 2R, 3S) l,2-O-isopropylidene-3-O-methyl-1,4-pentofuranos-4-yl])-propanoate. The following example broadly illustrates the nature of this invention, the manner in which it is to be performed without limiting the nature and scope of the invention. Example-1 Ethyl-3- (N- [N- (benzoyl), N- p-alanylamido-l-yl]-amino-3- [(1R, 2R, 3S) 1,2-0-isopropyIidene-3-0-methyI-l,4-pentofuranos-4-yI])-propanoate(7a): Sieber amide resin (1.1 gm, 0.68 mmol) was placed in solid phase reaction vessel and treated twice with 20% piperidine-DMF (10 mL each) for 5 min and 25 min at room temperature under N2 agitation. After this resin was successively washed with DMF (3x2 min), iPrOH (1x2 min) and DMF (3x2 min). Fmoc-(3- alanine was then added to the resin followed by addition of DIPEA (0.67 mL, 4.09 mmol), TBTu (.66 gm, 2.05 mmol)) and HOBt (0.31 gm, 2.05 mmol) in DMF (10 mL) with N2 agitation for 3 hrs at room temperature. Resin was washed with DMF (3 x 2min) and again treated with Fmoc-P- alanine in the presence of DIC (0.32 mL, 2.04 mmol) and HOBT (0.31 gm, 2.05 mmol) for 3 hrs. The resin was washed with DMF (3x2 min). Resin was washed with DMF (3 x 2min) and again treated with Fmoc-p- alanine in the presence of DIC (0.32 mL, 2.04 mmol) and HOBT (0.31 gm, 2.05 mmol) for 3 hrs. The resin was washed with DMF (3x2 min), MeOH (3x2 min) and DCMF (3x2 min). The completion of reaction was monitored by Kaiser test. The combinatorial library of glycoconjugates was synthesized in parallel format in the 96 well reaction block using 496 MOS Q automated synthesizer. P-alanyl acyl-Sieber amide resin (0.05 gm, 0.031 mmol) was placed in 24wells in the reaction block and treated with 20% piperidine-DMF (1 mL) twice for 5 min and 25 min. After this the resin in each well was washed with DMF (3x2 min), iPrOH (1x2 min) and DMF (3 x 2min). This was followed by transfer of ethyl- [3-(l,2-O-isopropylidene-3-O- methyl-1, 4-pentofuranos-4-yl]-propenoates to the desired well and subsequently treated with DBU (0.10 ml, 0.093 mmol) in DMF/ethanol for 48 hrs. After completion of the reaction the resin bound compound was again washed with DMF (3x2 min), iPrOH (1x2 min) and DMF (3 x 2min). The resin in each well was then treated with benzoyl chloride (0.15 ml) in presence of DBU (0.10 ml, 0.093 mmol) for 24 hrs. The completion of reaction was monitored by negative ninhydrin test. Finally the glycoconjugate was cleaved from the resin using mixture of 2% TFA, 94% dichloromethane and 4% triisopropylsilane. The solvent evaporated and the residue so obtained was dried over anhydrous KOH pellets overnight under reduced pressure. The product was then dissolved in t-Butanol/ Water and lyophilized to get the final compound as colorless solid. FAB MS:454[M+H]+. The glycoconjugate thus obtained was of 90% purity, as diastereoisomeric mixtures, which was sufficient for in vitro evaluation. Example-2 Ethyl-3- [N- (tetradecanoyl),N-( y-butyrylamido-1-yl)] -amino-3- [(1R, 2R, 3S) l,2-O-isopropylidene-3-0-methyI-l,4-pentofuranos-4-yl])-propionate(7b): Sieber amide resin (1.1 gm, 0.68 mmol) was placed in solid phase reaction vessel and treated twice with 20% piperidine-DMF (10 mL each) for 5 min and 25 min at room temperature under N2 agitation. After this resin was successively washed with DMF (3x2 min), iPrOH (1x2 min) and DMF (3x2 min). Fmoc-P- alanine was then added to the resin followed by addition of DIPEA (0.67 mL, 4.09 mmol), TBTu (.66 gm, 2.05 mmol)) and HOBt (0.31 gm, 2.05 mmol) in DMF (10 mL) with N2 agitation for 3 hrs at room temperature. Resin was washed with DMF (3 x 2min) and again treated with Fmoc-y- aminobutyric acid in the presence of DIC (0.32 mL, 2.04 mmol) and HOBT (0.31 gm, 2.05 mmol) for 3 hrs. The resin was washed with DMF (3x2 min), MeOH (3-x 2 min) and DCM (3x2 min). The completion of reaction was monitored by Kaiser test, y- aminobutanoyl-Sieber amide resin (0.05 gm, 0.031 mmol) was placed in the desired reaction block and treated with 20% piperidine-DMF (1 mL) twice for 5 min and 25 min. After this the resin in the well was washed with DMF (3x2 min), iPrOH (1x2 min) and DMF (3 x 2min). This was followed by transfer of the ethyl- [3-(l,2-0-isopropylidene-3-0-methyl)-l,4-pentofuranos-4-yl]-propenoates to the desired well and subsequently treated with DBU (0.10 mL, 0.093 mmol) in DMF/ethanol for 48 hrs. After completion the resins were again washed using the same wash chemfile as described earlier. The resin bound compound in well was then treated with the myristoyl chloride (0.25 ml) in pyridine (2ml) in presence of dimethyl aminopyridine and reaction continued for 24 hrs. The completion of reaction was monitored by negative ninhydrin test. Finally the glycoconjugate was cleaved from the resin using mixture of 2% TFA, 94% dichloromethane and 4%triisopropylsilane. The solvent evaporated and the residue dissolved in t-Butanol/ Water & lyophilized to get the desired compound. The compound wascharacterized using FAB MS: 586 [M+H]+. Example-3 Ethyl-3- [N- (octanoyl)- N- P-alanylamido-l-yl] -amino-3- [(1R, 2R, 3S) l,2-O-isopropyIidene-3-O-methyl-l, 4-pentofuranos-4-yl])-propanoate (7c): Sieber amide resin (1.1 gm, 0.68 mmol) was placed in solid phase reaction vessel and treated twice with 20% piperidine-DMF (10 mL each) for 5 min and 25 min at room temperature under N2 agitation. After this resin was successively washed with DMF (3 x 2 min), iPrOH (1x2 min) and DMF (3x2 min). Fmoc-P- alanine was then added to the resin followed by addition of DIPEA (0.67 mL, 4.09 mmol), TBTu (.66 gm, 2.05 mmol)) and HOBt (0.31 gm, 2.05 mmol) in DMF (10 mL) with N2 agitation for 3 hrs at room temperature. Resin was washed with DMF (3 x 2min) and again treated with Fmoc-p-alanine in the presence of DIC (0.32 mL, 2.04 mmol) and HOBT (0.31 gm, 2.05 mmol) for 3 hrs. The resin was washed with DMF (3x2 min). Resin was washed with DMF (3 x 2min) and again treated with Fmoc-P- alanine in the presence of DIC (0.32 mL, 2.04 mmol) and HOBT (0.31 gm, 2.05 mmol) for 3 hrs. The resin was washed with DMF (3 x 2 min), MeOH (3-x 2 min) and DCMF (3x2 min). The completion of reaction was monitored by Kaiser test, p-alanyl acyl-Sieber amide resin (0.05 gm, 0.031 mmol) was placed in the desired reaction block and treated with 20% piperidine-DMF (1 mL) twice for 5 min and 25 min. After this the resin in well was washed with DMF (3x2 min), iPrOH (1x2 min) and DMF (3 x 2min). This was followed by transfer of one of the ethyl- [3-(l,2-O-isopropylidene-3-O-methyl)-l,4-pentofuranos-4-yl]-propenoates to the desired well and subsequently treated with DBU (0.10 mL, 0.093 mmol) in DMF/ethanol for 48 hrs. After completion the resins were again washed using the same wash chemfile as described earlier. The resin in each well was then treated with the octanoyl chloride (0.25 ml) in pyridine (2ml) in presence of dimethyl aminopyridine and reaction continued for 24 hrs. The completion of reaction was monitored by negative ninhydrin test. Finally the glycoconjugate was cleaved from the resin using mixture of 2% TFA, 94% dichloromethane and 4%triisopropylsilane. The solvent evaporated and the residue dissolved in t-Butanol/ Water & lyophilized to get the desired compound. The compound was characterized using and FAB MS: 488 [M+H]+. Example-4 Ethyl-3- [N- dodecanoyl), N- a-amino butyryl-l-yl] -amino-3- [(1R, 2R, 3S) 1,2-0-isopropylidene-3-O-methyl-l, 4-pentofuranos-4-yl])-propanoate (7d): Sieber amide resin (1.1 gm, 0.68 mmol) was placed in solid phase reaction vessel and treated twice with 20% piperidine-DMF (10 mL each) for 5 min and 25 min at room temperature under N2 agitation. After this resin was successively washed with DMF (3 x 2 min), iPrOH (1x2 min) and DMF (3x2 min). Fmoc-a-amino butyric acid( 6 mmol) was then added to the resin followed by addition of DIPEA (0.67 mL, 4.09 mmol), TBTu (.66 gm, 2.05 mmol)) and HOBt (0.31 gm, 2.05 mmol) in DMF (10 mL) with N2 agitation for 3 hrs at room temperature. Resin was washed with DMF (3 x 2min) and again treated with Fmoc-a-amino butyric acid in the presence of DIC (0.32 mL, 2.04 mmol) and HOBT (0.31 gm, 2.05 mmol) for 3 hrs. The resin was washed with DMF (3x2 min). Resin was washed with DMF (3 x 2min) The resin was washed with DMF (3x2 min), MeOH (3-x 2 min) and DCM (3x2 min). The completion of reaction was monitored by Kaiser test. -a-amino butanoyl acyl-Sieber amide resin (0.05 gm, 0.031 mmol) was placed in the desired reaction block and treated with 20% piperidine-DMF (1 mL) twice for 5 min and 25 min. After this the resin in each well was washed with DMF (3x2 min), iPrOH (1x2 min) and DMF (3 x 2min). This was followed by transfer of the ethyl- [3-( 1,2-0-isopropylidene-3-O-methyl)-l,4-pentofuranos-4-yl]-propenoate to the desired well and subsequently treated with DBU (0.10 mL, 0.093 mmol) in DMF/ethanol for 48 hrs. After completion of the reaction the resins were again washed as above. The resin bound compound in reaction well was then treated with dodecanoyl chloride (0.15 ml) in DMSO (2ml) in presence of DBU (0.1 ml) and reaction continued for 24 hrs. The completion of reaction was monitored by negative ninhydrin test. Finally the glycoconjugate was cleaved from the resin using mixture of 2% TFA, 94% dichloromethane and 4% triisopropylsilane. The solvent evaporated and the residue dissolved in t-Butanol/ Water & lyophilized to get the desired compound. FAB MS: 530 [M+H]+. E-xample-5 Ethyl-3- [N- (hexadecanoyl), N- P-alanylamido-l-yl] -amino-3- [(1R, 2R, 3S) l,2-O-isopropylidene-3-O-methyl-l, 4-pentofuranos-4-yI])-propionate (7h): Sieber amide resin (1.1 gm, 0.68 mmol) was placed in solid phase reaction vessel and treated twice with 20% piperidine-DMF (10 mL each) for 5 min and 25 min at room temperature under N2 agitation. After this resin was successively washed with DMF (3x2 min), iPrOH (1x2 min) and DMF (3x2 min). Fmoc-P- alanine was then added to the resin followed by addition of DIPEA (0.67 mL, 4.09 mmol), TBTu (.66 gm, 2.05 mmol)) and HOBt (0.31 gm, 2.05 mmol) in DMF (10 mL) with N2 agitation for 3 hrs at room temperature. Resin was washed with DMF (3 x 2min) and again treated with Fmoc-P- alanine in the presence of DIC (0.32 mL, 2.04 mmol) and HOBT (0.31 gm, 2.05 mmol) for 3 hrs. The resin was washed with DMF (3x2 min). Resin was washed with DMF (3 x 2min) and again treated with Fmoc-p- alanine in the presence of DIC (0.32 mL, 2.04 mmol) and HOBT (0.31 gm, 2.05 mmol) for 3 hrs. The resin was washed with DMF (3x2 min), MeOH (3-x 2 min) and DCM (3x2 min). The completion of reaction was monitored by Kaiser test. P-Alanyl acyl-Sieber amide resin (0.05 gm, 0.031 mmol) was placed in s in the desired reaction block and treated with 20% piperidine-DMF (1 mL) twice for 5 min and 25 min. After this the resin in each well was washed with DMF (3x2 min), iPrOH (1x2 min) and DMF (3 x 2min). This was followed by transfer of the ethyl- [3-(l,2-O-isopropylidene-3-O-methyl)-l,4-pentofuranos-4-yl]-propenoate to the desired well and subsequently treated with DBU (0.10 mL, 0.093 mmol) in DMF/ethanol for 48 hrs. After completion the resins were again washed with DMF (3x2 min), MeOH (3-x 2 min) and DCM (3x2 min). The resin in each well was then treated with the palmitoyl chloride (0.15 ml) in pyridine (2ml) in presence of dimethyl aminopyridine (0.1 ml) and reaction continued for 24 hrs. The completion of reaction was monitored by negative ninhydrin test. Finally the glycoconjugate was cleaved from the resin using mixture of 2% TFA, 94% dichloromethane and 4%triisopropylsilane. The solvent evaporated and the residue dissolved in t-Butanol/ Water & lyophilized to get the desired compound. FAB MS : 572[M+H]+. ExampIe-6 Ethyl-3- [N- (dodecanoyl), N- (3-aIanylamido-l-yl] -amino-3- [(1R, 2R, 3S) l,2-O-isopropylidene-3-O-benzyI-l, 4-pentofuranos-4-yl])-propanoate (If): Sieber amide resin (1.1 gm, 0.68 mmol) was placed in solid phase reaction vessel and treated twice with 20% piperidine-DMF (10 mL each) for 5 min and 25 min at room temperature under N2 agitation. After this resin was successively washed with DMF (3x2 min), iPrOH (1x2 min) and DMF (3x2 min). Fmoc-P- alanine was then added to the resin followed by addition of DIPEA (0.67 mL, 4.09 mmol), TBTu (.66 gm, 2.05 mmol)) and HOBt (0.31 gm, 2.05 mmol) in DMF (10 mL) with N2 agitation for 3 hrs at room temperature. Resin was washed with DMF (3 x 2min) and again treated with Fmoc-P- alanine in the presence of DIC (0.32 mL, 2.04 mmol) and HOBT (0.31 gm, 2.05 mmol) for 3 hrs. The resin was washed with DMF (3 x 2 min). Resin was washed with DMF (3 x 2min) and again treated with Fmoc-P- alanine in the presence of DIC (0.32 mL, 2.04 mmol) and HOBT (0.31 gm, 2.05 mmol) for 3 hrs. The resin was washed with DMF (3x2 min), MeOH (3-x 2 min) and DCMF (3x2 min). The completion of reaction was monitored by Kaiser test. (3-alanyl acyl- Sieber amide resin (0.05 gm, 0.031 mmol) was placed in s in the desired reaction block and treated with 20% piperidine-DMF (1 mL) twice for 5 min and 25 min. After this the resin in each well was washed with DMF (3x2 min), iPrOH (1x2 min) and DMF (3 x 2min). This was followed by transfer of one of the ethyl- [3-(l,2-O-isopropylidene-3-O-methyl)-l,4- pentofuranos-4-yl]-propenoate to the desired well and subsequently treated with DBU (0.10 mL, 0.093 mmol) in DMF/ethanol for 48 hrs. After completion of the reaction the resin loaded compound were again washed as described earlier. The resin in each well was then treated with the dodecanoyl chloride (0.15 ml) in pyridine (2ml) in presence of dimethyl aminopyridine (0.1 ml) and reaction continued for 24 hrs. The completion of reaction was monitored by negative ninhydrin test. The resin was washed with DMF (3x2 min), iPrOH (1 x 2 min) and DMF (3 x 2mi.n). Finally the glycoconjugate was cleaved from the resin using mixture of 2% TFA, 94% dichloromethane and 4% triisopropylsilane. The solvent evaporated and the residue dissolved in t-butanol/ water & lyophilized to get the desired compound. The compound was characterized using FAB MS: 690 [M+H]+. Example-7 Ethyl-3- [N- (hexadecanoyl), N- P-alanylamido-l-yl] -amino-3- [(1R, 2R, 3S) l,2-O-isopropylidene-3-O-benzyl-l, 4-pentofuranos-4-yl])-propanoate (7e): Sieber amide resin (1.1 gm, 0.68 mmol) was placed in solid phase reaction vessel and treated twice with 20% piperidine-DMF (10 mL each) for 5 min and 25 min at room temperature under N2 agitation. After this resin was successively washed with DMF (3 x 2 min), iPrOH (1x2 min) and DMF (3x2 min). Fmoc-(3- alanine was then added to the resin followed by addition of DIPEA (0.67 mL, 4.09 mmol), TBTu (.66 gm, 2.05 mmol)) and HOBt (0.31 gm, 2.05 mmol) in DMF (10 mL) with N2 agitation for 3 hrs at room temperature. Resin was washed with DMF (3 x 2min) and again treated with Fmoc-P-alanine in the presence of DIC (0.32 mL, 2.04 mmol) and HOBT (0.31 gm, 2.05 mmol) for 3 hrs. The resin was washed with DMF (3x2 min), MeOH (3-x 2 min) and DCMF (3x2 min). The completion of reaction was monitored by Kaiser test. P-alanyl acyl-Sieber amide resin (0.05 gm, 0.031 mmol) was placed in s in the desired reaction block and treated with 20% piperidine-DMF (1 mL) twice for 5 min and 25 min. After this the resin in each well was washed with DMF (3x2 min), iPrOH (1x2 min) and DMF (3 x 2min). This was followed by transfer of the ethyl- [3-(l,2-O-isopropylidene-3-O-benzyl)-l,4-pentofuranos-4-yl]-propenoate to the desired well and subsequently treated with DBU (0.10 mL, 0.093 mmol) in DMF/ethanol for 48 hrs. After completion the resins were again washed as described earlier. The resin in reaction well was then treated with the hexadecanoyl chloride (0.15 ml) in pyridine (2ml) in presence of dimethyl aminopyridine (0.1 ml) and reaction continued for 24 hrs. The completion of reaction was monitored by negative ninhydrin test. Finally the glycoconjugate was cleaved from the resin using mixture of 2% TFA, 94% dichloromethane and 4%triisopropylsilane. The solvent evaporated and the residue dissolved in t-butanol/ water & lyophilized to get the desired compound. The compound was characterized using FAB MS : 732[M+H]+. Example-8 Ethyl-3- [N- (hexadecanoyl), N- p-alanylamido-l-yl] -amino-3- [(1R, 2R, 3S) l,2-O-isopropylidene-3-O-methyl-l, 4-pentofuranos-4-yl])-propanoate (7h): Sieber amide resin (1.1 gm, 0.68 mmol) was placed in solid phase reaction vessel and treated twice with 20% piperidine-DMF (10 mL each) for 5 min and 25 min at room temperature under N2 agitation. After this resin was successively washed with DMF (3x2 min), iPrOH (1x2 min) and DMF (3x2 min). Fmoc-p- alanine was then added to the resin followed by addition of DIPEA (0.67 mL, 4.09 mmol), TBTu (.66 gm, 2.05 mmol)) and HOBt (0.31 gm, 2.05 mmol) in DMF (10 mL) with N2 agitation for 3 hrs at room temperature. Resin was washed with DMF (3 x 2min) and again treated with Fmoc-p- alanine in the presence of DIC (0.32 mL, 2.04 mmol) and HOBT (0.31 gm, 2.05 mmol) for 3 hrs. The resin was washed with DMF (3x2 min), MeOH (3-x 2 min) and DCMF (3x2 min). The completion of reaction was monitored by Kaiser test. P-alanyl acyl-Sieber amide resin (0.05 gm, 0.031 mmol) was placed in s in the desired reaction block and treated with 20% piperidine-DMF (1 mL) twice for 5 min and 25 min. After this the resin in each well was washed with DMF (3 x 2 min), iPrOH (1x2 min) and DMF (3 x 2min). This was followed by transfer of the ethyl- [3-(l,2-0-isopropylidene-3-0-benzyl)-l,4-pentofuranos-4-yl]-propenoate to the desired well and subsequently treated with DBU (0.10 mL, 0.093 mmol) in DMF/ethanol for 48 hrs. After completion the resins were again washed as described earlier. The resin in reaction well was then treated with the hexadecanoyl chloride (0.15 ml) in pyridine (2ml) in presence of dimethyl aminopyridine (0.1 ml) and reaction continued for 24 hrs. The completion of reaction was monitored by negative ninhydrin test. Finally the glycoconjugate was cleaved from the resin using mixture of 2% TFA. 94% dichloromethane and 4%triisopropylsilane. The solvent evaporated and the residue dissolved in t-butanol/ water & lyophilized to get the desired compound. The compound was characterized using FAB MS : 657[M+H]+. Example- 9 The library of synthetic compounds was tested against commonest fungal pathogens Candid albicans and Cryptococcus neoformans by microbroth dilution technique (Iwata, K: Bossche, H.V.; In vitro and in vivo Evaluation of Antifungal Agents, Elesevier Science: Amsterdam, 1986, pp 31-34), in Sabouraud's dextrose broth at 28 °C. Stock solutions of the compounds were made in 10% DMSO (dimethyl sulfoxide) and the maximum concentration tested was 100 u.g/ml. The colony forming units (CFU) of both the organisms was maintained at lxlO4/ml in the test broth. The minimal inhibitory concentrations (MICs) were noted based upon absorbance value on an ELISA( Enzyme linked immunosorbent assay) reader at 492 nm after 24 hour (C.albicans) and 48 hour (C neoformans). The antifungal activity of some of the compounds from the library has been summarised in Table 1. Table 1: In vitro antifungal activity of compounds Table-1 Compound No. MICC.albocans Hg/ml C. neoformans 7a 100 100 7b >100 100 7c 25 50 7d 25 50 7f 25 100 7g 25 25 7h 100 100 Example 10 1. For MABA assay M. tuberculosis H37Ra was used as a suitable surrogate for the virulent H37 Rv strain. 7-8 days old cultures were diluted in liquid medium to provide an optimal density of 0.02 at 550 nm spectrophotometrically which gave colour change of Alamar blue 'oxidation reduction' dye (blue to pink). A compound with MIC Table-2: In vitro antitubercular activity Compounds MIC(ng /ml) 7b 12.5 7d 12.5 7g 12.5 Rifamycin 0.005 Isoniazid 0.75 Example- 11 The activity of compounds was tested against bioluminiscent Mycobacterium aurum expressing_firefly luciferase (D.K.Deb; K.K.Srivastava; Ranjana Srivastava; and B.S. Srivastava Biochem. Biophys. Res. Comm. 279, 457-61, 2000). The cells were grown to an optical density of 0.03 at 600 nm. Two fold dilutions of compounds were prepared and added to IOOJJI culture (Agoo = 0.03) in microtitre plate. The plate was incubated at 37 °C for 6 hr and bioluminiscence was measured for each well. Two controls (with no drug) and two standard drugs (rifampicin and sparfloxacin) were also included. For measurement of bioluminescence IOOJJ.1 of culture was mixed with 250 \xl of sodium citrate buffer (0.1M, pH 5.0) in the tube and was placed in the luminometer. (Lumat LB 9507- EG & G Berthhold) lOOu.1 of 1 mM luciferase substrate was infected and luminescence was measured as relative light units (RLU) for 10s. The antitubercular activities of the compounds are given in Table-3 and Table-4. Table-3: In vitro antitubercular activity Treatment Concentration ((^g /ml) RLU _____ . 1.3 x io6 Rifampicin 2.0 4xlO3 Sparfloxacin 0.5 1.1 x 103 7e 12.5 2xlO4 7f 12.5 1.5 xlO4 7g 12.5 3.3 x 104 7h 12.5 3.1 x 104 TabIe-4:1 n vitro antitubercular activity Compound/drug MIC (jig/ml) 7b 12l 7c 12.5 7j 12.5 Rifamycin 0.005 Isoniazid 0.75 Table-S: In vitro antitubercular activity Compound no MIC (ng/ml) 7a 25 7d 12.5 7f 25 7h 50 7i 12.5 Example-12 In macrophage method of screening the combinatorial library the macrophage cell line J744A/ has been cultured in DMEM (Dulbecos modified Eagles medium) medium with 10% FCS at 37 ° C in the presence of 5% CO2. The cultures were infected with 10 m.o.i. of M aurum (grown in sautan medium). After 5hr extracellular bacilli were washed out and the adherent macrophage cells containing mycobacterium were replenished with fresh medium. The drugs were added in different concentration and the infected cultures were reincubated -25 for a period ranging from 24 to 48 h. After this period the infected macrophages were lysed and plated on nutrient agar tween plates for colony forming units. The data were compared with the untreated samples. In vitro antitubercular activity is given in Table-5 We claim 1. A combinatorial library of 3-substituted amino-3-glycosylated propanoates useful as antifungal and antibacterial agents of the general formula 7 (figure-1) (Formula Removed) wherein R is the alkyl chain consisting of 1-4 carbon alkyl, aralkyl groups or H, R1 is alkyl group or H; R2 is 1-4 carbon branched or unbranched alkyl groups; R3 is branched or unbranched alkyl, heterocycloalkyl or cyclogroup or H; n ranges from 0 to 4, R" is 1 to 4 carbon branched or unbranched alkyl or acyl group, cycloalkyl or heterocycloalkyl groups, aroyl group where aromatic ring is substituted or unsubstituted. 2. A combinatorial library of 3-substituted amino-3-glycosylated propanoates propanoates useful as antifungal and antibacterial agents as claimed in claim 1, wherein the representative compounds are as follows Ethyl-3- [N- (tetradecanoyl), N- -butyralamido- 1 -yl] -amino-3- 1(1R, 2R, 3S) 1,2- 0-isopropylidene-3-0-methyl- 1, 4-pentofuranos-4-ylp-propanoate, Ethyl-3- [N- (octanoyl), N- p-alanylamido-1 -yll -amino-3- RIR 2R, 38) 1, 2-0- sopropylidene-3-0-methy 1-1, 4-pentofuranos-4-y lD-propanoate, Ethyl-3- [N- (dodecanoyl), N-(a-aminobutyralmido)-l-yl] -amino-3- RJR, 2R, 35) 1,2-0-lsopropylidene-3-0-methyl-l, 4-pentofuranos-4-ylp-propanoate, Ethyl-3-[N- (hexadecanoyl), N- P-alanylamido-1-yl] -amino-3- RJR, 2R, 38) 1, 2-0- isopropylidene-3-0-benzy 1 -1,4-pentofuranos-4-y 1D-propanoate, Ethyl-3-[N- (dodecanoyl), N-p-alanylamido-1-yl] -amino-3- [(IR, 2R, 35) 1,2-0- isopropylidene-3-0-benzy 1 -1, 4-pentofuranos-4-ylp-propanoate, Ethyl-3-[N- (hexadecanoyl),N-p-alanylamido-l-yl] -amino-3- [(IR, 2R, 35) 1,2-0-isopropyl idene-3-0-benzy 1-1, 4-pentofuranos-4-yl])-propanoate, Ethyl-3-[N-(hexadecanoyl),N-p-alanylamido-l-yl] -amino-3- [(IR, 2R, 3S) 1,2-0-isopropylidene-3-0-methyl-l, 4-pentofuranos-4-yl])-propanoate, Ethyl-3-[N-(tetradecyl),N-p-alanylamido-l-yl]-amino-3-[(IR,2R,3S)1,2-0-isopropylidene-3-0-methyl-l,4-pentofuranos-4-yl])-propanoate, Ethyl-3-[N-(dodecanoyl),N-p-alanylamido-l-yl]-amino-3-[(1R,2R,3S)l,2-0-isopropylidene-3-0-methy 1-1, 4-pento furanos-4-ylp-propanoate, Ethyl-3-[N-(benzoyl),N-p-alanylamido-l-yl]-amino-3-[(IR,2R,3S)l,2-0-isopropylidene-3-0-methy 1-1, 4-pentofuranos-4-yl])-propanoate, Ethyl-3-[N-(benzoyl),N-p-alanylamido-l-yl]-amino-3-[(IR,2R,3S)1,2-0-isopropylidene-3-0-benzyl-l, 4-pentofuranos-4-ylp-propanoate, Ethyl-3-[N- (benzyl),N-p-alanylamido-l-yl] -amino-3- [(IR, 2R, 3S) 1,2-0- isopropyl idene-3 -0-methyl-1, 4-pentofuranos-4-yll)-propanoate, Ethyl-3-[N-(4-cyanobenzyl),N-p-alanylamido-1 -yl]-amino-3-[(IR,2R,3S) 1,2-0-isopropylidene-3-0-methy 1-1, 4-pentofuranos-4-yl])-propanoat, Ethyl-3-[N-(dodecanoyl),N-(hexamido-6-yl)]-amino-3-[(IR,2R,3S) 1,2-0-isopropylidene-3-0-methy 1-1, 4-pentofuranos-4-y 1 l)-propanoate, Ethyl-3-[N-(octanoyl),N-(hexamido-6-yl)-amino-3- [(IR,2R, 3S) 1,2-0- isopropylidene- 3-0-methy 1 -1, 4-pentofuranos-4-y 1])-propanoate, Ethyl-3-[N-(benzoyl),N-(phenylalanylamido-2-yl)]-amino-3-[(IR,2R,3S) 1,2-0- isopropylidene- 3-0-methyl- I, 4-pentofuranos-4-yl])-propanoate, Ethyl-3- [N- (benzyl), N-(phenylalanylamido-2-yl)] -amino-3- [(IR,2R,3S) 1, 2-0- isopropylidene- 3-0-methyl- I, 4-pentofuranos-4-yl])-propanoate, Ethyl-3-[N- (benzyl), N-(phenylalanylamido-2-yl)]-amino-3-[(IR,2R,3S)l,2-0- isopropy lidene- 3-0-benzyl-l, 4-pentofuranos-4-yl])-propanoate, Ethyl-3- [N- (4-cyanobenzyl), N-(phenylalanylamido-2-yl)l -amino-3- [(1R, 2R. 3S) 1,2-0- isopropylidene-3-0-methy 1-1, 4-pentofuranos-4-yl])-propanoate, Ethyl-3- [N- (benzyl), N-(propanamido-2-yl)] -amino-3- [{IR, 2R, 3S) 1,2-0- isopropylidene- 3-0-methyl-l, 4-pentofuranos-4-yl])-propanoate, Ethyl-3- [N- (benzoyl), N-(propanamido-2-yl)] -amino-3-[(IR,2R,3S) 1,2-0- isopropylidene- 3-0-methyl-l, 4-pentofuranos-4-yl])-propanoate, Ethyl-3- [N-(dodecanoyl),N-(propanamido-2-yl)] -amino-3- [(IR, 2R 3S) 1,2-0- isopropylidene- 3-0-methyl-l, 4-pentofuranos-4-yl])-propanoate, Ethyl-3-[N-(tetrdecanoyl),N-(propanamido-2-yl)]-amino-3- [(IR, 2R, 3S)l,2-0- isopropylidene- 3-0-methyl- I, 4-pentofuranos-4-yl])-propanoate, Ethyl-3 - [N- (pentaEthyl-3- [N- (tetrdecanoyl), N-(propanamido-2-yl)] -am i no-3 - [(1R, 2R, 3S) l,2-0-isopropylidene-3-0-methyl-l, 4-pentofuranos-4-ylD-propanoate decanoyl), N- (propanamido-2-y1)] -amino-3-[(7R, 2R,35') l,2-0-isopropylidene-3-0-methyl- I, 4- pentofuranos-4-yl])-propanoate. 3. A combinatorial library of 3-substituted amino-3-glycosylated propanoates useful as antifungal and antibacterial agents substantially as herein described with reference to the examples and drawing accompanying the specifications.

Full Text

The present invention relates to a novel combinatorial library of 3-substituted amino-3-glycosylated propanoate derivatives. More particularly the present invention relates to a novel combinatorial library of 3-substituted amino-3-glycosylated propanoate derivatives useful as antifungal and antibacterial agents. Background of the invention:
Efficiency has been the key goal in modern organic synthesis of biomolecules in drug design and discovery. Combinatorial synthesis is a method to prepare a large number of structurally distinct molecules where many experiments may be carried out at the same time. It has been achieved both by solution and solid phase methods using either divide & couple or split techniques (A, Netzi, J.M. Ostresh, R.A. Houghten Chem Rev 1997. 97, 449). The parallel synthesis of numerous single compounds is considered to be most desirable nowadays and can be accomplished by sequential coupling to solid phase linked substrate if suitable combination of protecting and anchoring groups are applied. In comparison to other scaffolds carbohydrate derived scaffolds constitute template, which not only offer more utilizable functional groups but also numerous, alternatively addressable stereogenic centers (T. Wunberg, C. Kallus, T. Opatz, S. Henko, W. Schimdt, H. Kunz Angew. Chem. Int. Ed. 1998, 37, 2503).
Despite the availability of effective and short course chemotherapy and BCG vaccine the tube -rcular baccilus continues to claim more lives than any other single infectious agent in the world. A number of substances other than the antibiotics have been proposed heretofore for use in treating tubercular and fungal infections, but they have not been used extensively for a number of reasons. Among them the increased incidence of multidrug resistant tuberculosis, its deadly synergy with human immunodeficiency virus and susceptibility to fungal infections

in tuberculosis and resistance to existing antifungals are the prominent ones. Many potentially effective antitubercular and antifungal agents are toxic and cannot be safely administered to patients and still some of them have undesirable side effects. As a result of the foregoing and other deficiencies, it is apparent that the development of new class of nontoxic mechanism based broad spectrum antimicrobial agents has not been available for treating the tubercular and fungal infections.
It has been discovered that certain glycosylated amino acids are associated with a number of biological activities as these provide important biological signals to the cells in order to combat the adverse conditions (Varki A., Glycobiology (1993), 3, 97 & Drickamen, K. In molecular glycobiology Flukada, M., Hindsgaul. O., Eds IRL, Oxford 1994, Ch2, pp53-87). Further sugar derivatives are known to offer better pharmacokinetic parameter to the drugs and at the same time they are less toxic in comparision to other class of compounds [J.F.Fisher, Harrison, A.W., Bundy G.L., K.F. Willinkson, Rush B.D., Ruwart M.J., J. Med. Chem(1991, 34, 3410 & Negre E., Driaf K.P., Veraneuil B, Tetrahedron Lett (1993), 34, 1027]. Combinatorial synthesis of small molecules has already gained momentum in drug discovery and research. However, the combinatorial syntheses of glycoconjugate have been very slow because of difficulty in their synthesis and structure elucidation. Further, glycoconjugates are known to be superior ligands with higher affinity than a natural carbohydrate ligand [P. M. St. Hilaire and M. Meldal Angew. Chem. Int. Ed (2000, 39, 1162)]. Hence, these carbohydrate based libraries can be essential tools in the development of carbohydrate based therapeutics particularly the anti-infectives.
The main objective of the present invention is to provide novel combinatorial library of 3-substituted amino-3-glycosylated propanoate derivatives useful as antifungal and antibacterial agents.

Another objective of the present invention is to provide a process for preparation of novel combinatorial library of 3-substituted amino-3-glycosylated propanoate derivatives of the general formula 7 (Figure-1).
Yet another objective of the present invention is to provide compounds useful in the treatment of fungal and mycobacterial infections.
Accordingly the present invention relates to novel combinatorial library of 3-substituted amino-3-glycosylated propanoates of the general formula 7 (figure-1)
H2NC(CH2)nCHN

wherein R is the alkyl chain consisting of 1-4 carbon alkyl, aralkyl groups or H, R| is alkyl group or H; R2 is 1-4 carbon branched or unbranched alkyl groups; R3 is branched or unbranched alkyl, heterocycloalkyl or cyclogroup or H; n ranges from 0 to 4, R4 is 1 to 4 carbon branched or unbranched alkyl or acyl group , cycloalkyl or heterocycloalkyl groups, aroyl group where aromatic ring is substituted or unsubstituted; which comprises (i) preparing activated sieber amide resin by treating the sieber amide resin with 20-90 % piperidine-dimethyl formamide or piperidine, (ii) coupling the activated amide resin with Fmoc-aminoacids of the general formula 1, wherein R3 is branched or unbranched alkyl, heterocycloalkyl or cycloalkyl group or H; n ranges from 0 to 4, in presence of conventional coupling agent at a temperature in the range of 0-100 °C for a period in the range of 1 to 12 hrs to form amide linkage between the resin and amino acid of the general formula 1 producing the resin bound compound of the general formula 2 wherin value of R3 and n is same as given for

formula 1, (iii) deprotecting the Fmoc group from the above substrate 2, by treating with 20-90 % piperidine in dimethyl formamide at 0 to 50 °C, followed by washing with dimethyl formamide and successively with dichloromethane to get the free amino group resin bound substrate, (iv)reacting alkyl- [3-(l,2-O-isopropylidene-3-O-substituted (alkyl or aralkyl)-l,4-pentofuranos-4-yl]-propenoates of the general formula 3 (figure-1) wherein R is the alkyl chain consisting of 1-4 carbon alkyl , aralkyl groups or H, Ri is alkyl group or H; R2 is 1-4 carbon branched or unbranched alkyl groups; to the aminoacyl resin in an organic solvent in presence of an organic base at the temperature in the range of 10-100 °C for a period 1-48 hrs, washing the resin bound compound of formula 4 wherein R, Rl and R2 are same as given for formula 2 and 3 as in step (iii); (v) reacting the resin bound compound of the general formula 4 either with a compound selected from alkyl or arylhalides/ acyl halides/ aliphatic / aromatic/ heteroaromatic acids or their acid chlorides of the general formula 5, wherein R4 is 1-12 carbon branched or unbranched alkyl chain , substituted or unsubstituted aromatic/heteroaromatic rings , X is CH2CI, CI-bBr , COC1 , COBr or COOH groups, n may vary from 0 to 12;, in dimethyl formamide/ dimethyl sulphoxide in presence of bases selected from diazabicycloundecene, pyridine, dimethyl amino pyridine at temperature ranging from 0 to 150 °C for a period 1-56 hrs and washing the reaction mixture as in case of step (iii) to give the resin bound compound of the general formula 6 wherein R, Ri, R2, R3 and R4 as stated above finally cleaving the resin bound glycoconjugates to give the compounds of the general formula 7 using 2-90% trifluoro acetic acid in dichloromethane in the temperature range of 0 to 40 °C; evaporating the solvents followed by lyophilization using tertiary butanol and water.

The present invention provide a novel combinatorial library of 3-substituted- amino-3-glycosylated propanoates useful as antifiingal and antibacterial agents which comprises the following steps; (a) swelling of solid phase (sieber amide resin) with solvents such as dicloromethane, dimethyl formamide in nitrogen atmosphere; (b) Deprotection of Fmoc-group of the sieber amide resin to corresponding free amino sieber amide using 30-90% piperidine-DMF (dimethyl formamide); purging the excess solvents and washing the excessive reagents; (c) loading of the Fmoc-NH-amino acids of the general formula 1, on to a solid phase such as sieber amide resin using HOBt (1-hydroxy benzotriazole), DIPEA (di-isopropyl ethyl amine), TBTU [2-(lH- benzotriazole-1-yl) 1,1,3,3-tetramethyl uroniumtetrafluoroborate], DIPC (diisopropyl carbodimide) in an organic solvent at a temperaure in the range of 0-100 °C for a period ranging from 1 to 12 hrs; purging the excess solvents and washing the excessive reagents to give the resin bound compound of formula 2; (d) deprotecting the Fmoc group in compound 2 with 30-90% piperidine-DMF followed by washing with isopropanol and dichloromethane; (d) reacting alkyl- [3-(l,2-O-isopropylidene-3-O-substituted (alkyl or aralkyl)-l,4-pentofuranos-4-yl]-propenoates (R.P.Tripathi et.al. Indian Patent application No 48/DEL/2001) of the general formula 3 (figure-1) to the Fmoc deprotected aminoacyl resin of the general formula 2 in an organic solvent in presence of DBU (diazabicycloundecene) or triethyl amine at the temperature in the range of 10-100 °C for a period 1-48 hrs, purging the excess solvents and washing the excessive reagents to give the resin bound compound of the general formula 4; and (e) Functionalising secondary amino group with alkanoic acids, alkyl halides, acyl chlorides, hetero aromatic acids; of general formula 5 and 6 in an organic solvent at a temperature in the range of 0° C to 150° C for a period in the range of 1 to 56 hr, in presence of bases such as diazabicycloundecene, pyridine,

dimethyl amino pyridine; purging the excess solvents and washing the excessive reagents to get the resin bound compound of the general formula 7; and (f) by cleaving the resin bound glycoconjugates using 2-50 % trifluoro acetic acid in dichloromethane at the temperature in the range of 0-40 ° C; evaporating the solvents at temperature 0-100 °C followed by freeze drying of the products so obtained to give the combinatorial library of glycoconjugates of general formula 7 (figure-1).
In an embodiment of the present invention the reaction vessels used may be selected from syringes, automatecd robotic synthesiser, Merrifield's vessel, three way sintered vessel.
In an embodiment of the present invention the solid phase may be selected from super acid sensitive resins sieber amide or rink amide resins.
In another embodiment of the present invention the solvents used for swelling the resins may be selected from dicloromethane, dimethyl formamide and tetrahydrofuran.
In yet another embodiment of the present invention the inert atmosphere used may be selected from nitrogen or argon atmosphere.
In a further embodiment of the present invention the reagents used for deblocking the Fmoc group from the resins as well as resin bound compounds may be selected from 30-100 % piperidine in dimethyl formamide, triisopropyl silane, triethyl amine.
In yet another embodiment of the present invention the solvent used in the reaction of Fmoc amino glycosylated propanoic acids and activated sieber amide resin, amino glycosylated propanoyl resin and Fmoc amino acid, aromatic or heteroaromatic acids may be selected from dimethyl formamide, dichloromethane, tetrahydrofuran.
In yet another embodiment of the present invention the solvent used in the reaction of olefinic ester derived from sugars and resin bound amino acyl derivatives may be selected

from alcohols such as methanol, ethanol, propanol, dimethyl formamide, tetrhydrofiiran, dioxane, dichloromethane etc.
In a further embodiment of the present invention the catalyst used in the reaction between olefinic ester derived from sugars and resin bound amino acyl derivatives may be selected from triethyl amine, p-dimethyl amino pyridine, diazabicycloundecene or phase transefer catalyst such as tetra butyl ammonium bromide, Tetramethyl ammonium hydroxide, tetrabenzyl ammonium bromide.
In a further embodiment of the present invention the reagent used in cleaving the compounds loaded on to resins may be 2-50% trifluoroacetic acid in dichloromethane.
Thus the process used for this invention comprises the following steps (i) Sieber amide resin was placed in 24 wells in the reaction block of robotic synthesizer and treated with 20% piperidine-DMF (1 ml) twice for 5 min and 25 min. After this the resin in each well was washed with DMF (3x2 min), iPrOH (1x2 min) and DMF (3 x 2min). This was followed by transfer of one of the Fmoc amino acids such as P-alanine, leucine, isoleucine, a-amino butyric acid, y-amino butyric acid, valine or nor valine or their derivatives to the desired well and subsequently treated with, TBTU, DIC and HOBt in DMF and the reaction was carried out for 24 hrs. After completion the resins were again washed with DMF, dichloromethane, isopropanol separately. (iii)The Fmoc protecting group was deblocked using 20 % piperidine-DMF and washed with DMF, dichloromethane and solvent purged off and then treated with the desired alkyl- [3-(l,2-O-isopropylidene-3-O-substituted (alkyl or aralkyl)-l,4-pentofuranos-4-yl]-propionates in DMF and ethanol (1:1) in presence of triethyl amine or diazabicycloundecene (DBU) and the reaction wells were stirred for 24 hrs and the reaction vessels washed with DMF (3x2 min), iPrOH (1x2 min) and DMF (3 x 2min). The completion of reaction was monitored by negative ninhydrin test. (iv)This was followed by

removal of Fmoc group in each well with 20% piperidine-DMF and washing the reaction vessels with DMF (3x2 min), iPrOH (1x2 min) and DMF (3 x 2min), drying the reaction wells. (v)The resin bound compounds in each well was treated separately with different alkyl halides, araalkyl halides or acyl halides in pyridine, dimethyl sulphoxide, DMF, THF etc in presence of bases such a triethyl amine, diazabicycloundecene, pyridine at 0 to 100 ° C and the reaction wells stirred for 20 hrs. The solvent and unreacted material was purged off. The reaction wells were washed with DMF (3x2 min), iPrOH (1x2 min) and DMF (3 x 2min) Finally the glycoconjugates were cleaved from the resin using mixture of 2% TFA, 94% dichloromethane and 4% triisopropylsilane. Evaporating the solvents and dissolving the products, so obtained, in t-Butanol/ Water (4:1) and lyophilizing the final compounds after deprotection and cleavage.
In our copending patent application no 1272/del/01, claimed and disclosed the process for the preparation of novel combinatorial library of 3-substituted amino-3-glycosylated propanoates. From this application we have divided out this application The representative compounds of the formula 7 prepared by the process of the present invention are as follows:
Ethyl-3- (N- [N- (benzoyl), N- p-alanylamido-l-yl]-amino-3- [(1R, 2R, 3S) 1,2-0-
isopropylidene-3-O-methyl-1,4-pentofuranos-4-yl])-propanoate
Ethyl-3- [N- (4-Nitrobenzoyl), N- (3-alanylamido-l-yl] -amino-3- [(1R, 2R, 3S) 1,2-0-
isopropylidene-3-O-methyl-l, 4-pentofuranos-4-yl])-propanoate
Ethyl-3- [N- (tetradecanoyl), N- p-alanylamido-1-yl] -amino-3- [(1R, 2R, 3S) 1,2-0-
isopropylidene-3-O-benzyl-1,4-pentofuranos-4-yl])-propanoate

Ethyl-3- [N- (dodecanoyl), N- P-alanylamido-1-yl] -amino-3- [(1R, 2R, 3S) 1,2-0-Isopropylidene-3-O-benzyl-1,4-pentofuranos-4-yl])-propanoate Ethyl-3- [N- (hexadecanoyl), N- P-alanylamido-1-yl] -amino-3- [(1R, 2R, 3S) 1,2-0-isopropylidene-3-O-benzyl-1,4-pentofuranos-4-yl])-propanoate Ethyl-3- [N- (dodecanoyl), N- p-alanylamido-1-yl] -amino-3- [(1R, 2R, 3S) l,2-O-isopropylidene-3 -O-methyl-1,4-pentofuranos-4-yl])-propanoate Ethyl-3- [N- (hexadecanoyl), N- p-alanylamido-1-yl] -amino-3- [(1R, 2R, 3S) 1,2-0-isopropylidene-3-O-benzyl-l, 4-pentofiiranos-4-yl])-propanoate Ethyl-3- [N- (hexadecanoyl), N- P-alanylamido-1-yl] -amino-3- [(1R, 2R, 3S) 1,2-0-isopropylidene-3-O-methyl-1,4-pentofuranos-4-yl])-propanoate Ethyl-3- [N- (tetradecanoyl), N-y-butyralamido-1-yl] -amino-3- [(1R, 2R, 3S) 1,2-O- isopropylidene-3-O-methyl-1, 4-pentofiiranos-4-yl])-propanoate Ethyl-3- [N- (octanoyl), N- p-alanylamido-1-yl] -amino-3- [(1R, 2R, 3S) 1,2-0-sopropylidene-3-O-methyl-1,4-pentofuranos-4-yl])-propanoate Ethyl-3- [N- (dodecanoyl), N-(a-aminobutyralmido)-l-yl] -amino-3- [{1R, 2R, 3S) l,2-O-Isopropylidene-3-O-methyl-l,4-pentofiiranos-4-yl])-propanoate Ethyl-3- [N- (hexadecanoyl), N- p-alanylamido-1-yl] -amino-3- [(1R, 2R, 3S) 1,2-0-isopropylidene-3-O-benzyl-l, 4-pentofiiranos-4-yl])-propanoate Ethyl-3- [N- (dodecanoyl), N- P-alanylamido-1-yl] -amino-3- [(1R, 2R, 3S) 1,2-0-isopropylidene-3-O-benzyl-1,4-pentofuranos-4-yl])-propanoate Ethyl-3- [N- (hexadecanoyl), N- p-alanylamido-1-yl] -amino-3- [(1R, 2R, 3S) 1,2-0-isopropylidene-3-O-benzyl-1,4-pentofuranos-4-yl])-propanoate

Ethyl-3- [N- (hexadecanoyl), N- (3-alanylamido-l-yl] -amino-3- [(1R, 2R, 35) 1,2-0-
isopropylidene-3-O-methyl-1,4-pentofiiranos-4-yl])-propanoate
Ethyl-3- [N- (tetradecyl), N- P-alanylamido-1-yl] -amino-3- [(1R, 2R, 35) l,2-O-
isopropylidene-3-O-methyl-1,4-pentofuranos-4-yl])-propanoate
Ethyl-3- [N- (dodecanoyl), N- p-alanylamido-1-yl] -amino-3- [(1R, 2R, 35) 1,2-0-
isopropylidene-3-O-methyl-1,4-pentofuranos-4-yl])-propanoate
Ethyl-3- [N- (benzoyl), N- P-alanylamido-1-yl] -amino-3- [(1R, 2R, 35) 1,2-0-
isopropylidene-3-O-methyl-1,4-pentofuranos-4-yl])-propanoate
Ethyl-3- [N- (benzoyl), N- p-alanylamido-1-yl] -amino-3- [(1R, 2R, 3S) 1,2-0-
isopropylidene-3-O-benzyl-l,4-pentofuranos-4-yl])-propanoate,
Ethyl-3- [N- (benzyl), N- p-alanylamido-1-yl] -amino-3- [(1R, 2R, 3S) 1,2-0-
isopropylidene-3-O-methyl-1,4-pentofiiranos-4-yl])-propanoate
Ethyl-3- [N- (4-cyanobenzyl), N- P-alanylamido-1-yl] -amino-3- [(1R, 2R, 35) 1,2-0-
isopropylidene-3-O-methyl-l, 4-pentofuranos-4-yl])-propanoate
Ethyl-3- [N- (dodecanoyl), N-(hexamido-6-yl)] -amino-3- [(1R, 2R, 35) 1,2-0-
isopropylidene-3-O-methyl-l, 4-pentofuranos-4-yl])-propanoate
Ethyl-3- [N- (octanoyl), N-(hexamido-6-yl)] -amino-3- [(1R, 2R, 35) 1,2-0-
isopropylidene-3-O-methyl-1,4-pentofuranos-4-yl])-propanoate
Ethyl-3- [N- (benzoyl), N-(phenylalanylamido-2-yl)] -amino-3- [(1R, 2R, 35) 1,2-0-
isopropylidene-3-O-methyl-l, 4-pentofuranos-4-yl])-propanoate
Ethyl-3- [N- (benzyl), N-(phenylalanylamido-2-yl)] -amino-3- [(1R, 2R, 35) l,2-O-
isopropylidene-3-O-methyl-l, 4-pentofUranos-4-yl])-propanoate

Ethyl-3- [N- (benzyl), N-(phenylalanylamido-2-yl)] -amino-3- [(1R, 2R, 3S) 1,2-0-
isopropylidene-3-O-benzyl-l, 4-pentofiiranos-4-yl])-propanoate
Ethyl-3- [N- (4-cyanobenzyl), N-(phenylalanylamido-2-yl)] -amino-3- [(1R, 2R, 3S)
1,2-O-isopropylidene-3-O-methyl-1, 4-pentofuranos-4-yl])-propanoate
Ethyl-3- [N- (benzyl), N-(propanamido-2-yl)] -amino-3- [(1R, 2R, 3S) 1,2-0-
isopropylidene-3-O-methyl-1,4-pentofiiranos-4-yl])-propanoate
Ethyl-3- [N- (benzoyl), N-(propanamido-2-yl)] -amino-3- [(1R, 2R, 3S) 1,2-0-
isopropylidene-3-O-methyl-1,4-pentofuranos-4-yl])-propanoate
Ethyl-3- [N- (dodecanoyl), N-(propanamido-2-yl)] -amino-3- [(1R, 2R, 3S) 1,2-0-
isopropylidene-3-O-methyl-l, 4-pentofuranos-4-yl])-propanoate
Ethyl-3- [N- (tetrdecanoyl), N-(propanamido-2-yl)] -amino-3- [(1R, 2R, 3S) 1,2-0-
isopropylidene-3-O-methyl-1,4-pentofuranos-4-yl])-propanoate
Ethyl-3- [N- (pentaEthyl-3- [N- (tetrdecanoyl), N-(propanamido-2-yl)] -amino-3- [(1R, 2R, 3S) l,2-O-isopropylidene-3-O-methyl-l, 4-pentofuranos-4-yl])-propanoate decanoyl), N-(propanamido-2-yl)] -amino-3- [{1R, 2R, 3S) l,2-O-isopropylidene-3-O-methyl-1,4-pentofuranos-4-yl])-propanoate.
The following example broadly illustrates the nature of this invention, the manner in which it is to be performed without limiting the nature and scope of the invention.
Example-1
Ethyl-3- (N- [N- (benzoyl), N- p-alanylamido-l-yl]-amino-3- [(1R, 2R, 3S) 1,2-0-isopropyIidene-3-0-methyI-l,4-pentofuranos-4-yI])-propanoate(7a):
Sieber amide resin (1.1 gm, 0.68 mmol) was placed in solid phase reaction vessel and treated twice with 20% piperidine-DMF (10 mL each) for 5 min and 25 min at room temperature under N2 agitation. After this resin was successively washed with DMF (3x2 min), iPrOH (1x2 min) and DMF (3x2 min). Fmoc-(3- alanine was then added to the resin

followed by addition of DIPEA (0.67 mL, 4.09 mmol), TBTu (.66 gm, 2.05 mmol)) and HOBt (0.31 gm, 2.05 mmol) in DMF (10 mL) with N2 agitation for 3 hrs at room temperature. Resin was washed with DMF (3 x 2min) and again treated with Fmoc-P- alanine in the presence of DIC (0.32 mL, 2.04 mmol) and HOBT (0.31 gm, 2.05 mmol) for 3 hrs. The resin was washed with DMF (3x2 min). Resin was washed with DMF (3 x 2min) and again treated with Fmoc-p- alanine in the presence of DIC (0.32 mL, 2.04 mmol) and HOBT (0.31 gm, 2.05 mmol) for 3 hrs. The resin was washed with DMF (3x2 min), MeOH (3x2 min) and DCMF (3x2 min). The completion of reaction was monitored by Kaiser test. The combinatorial library of glycoconjugates was synthesized in parallel format in the 96 well reaction block using 496 MOS Q automated synthesizer. P-alanyl acyl-Sieber amide resin (0.05 gm, 0.031 mmol) was placed in 24wells in the reaction block and treated with 20% piperidine-DMF (1 mL) twice for 5 min and 25 min. After this the resin in each well was washed with DMF (3x2 min), iPrOH (1x2 min) and DMF (3 x 2min). This was followed by transfer of ethyl- [3-(l,2-O-isopropylidene-3-O- methyl-1, 4-pentofuranos-4-yl]-propenoates to the desired well and subsequently treated with DBU (0.10 ml, 0.093 mmol) in DMF/ethanol for 48 hrs. After completion of the reaction the resin bound compound was again washed with DMF (3x2 min), iPrOH (1x2 min) and DMF (3 x 2min). The resin in each well was then treated with benzoyl chloride (0.15 ml) in presence of DBU (0.10 ml, 0.093 mmol) for 24 hrs. The completion of reaction was monitored by negative ninhydrin test. Finally the glycoconjugate was cleaved from the resin using mixture of 2% TFA, 94% dichloromethane and 4% triisopropylsilane. The solvent evaporated and the residue so obtained was dried over anhydrous KOH pellets overnight under reduced pressure. The product was then dissolved in t-Butanol/ Water and lyophilized to get the final compound as colorless solid. FAB MS:454[M+H]+. The glycoconjugate thus obtained was of 90% purity, as diastereoisomeric mixtures, which was sufficient for in vitro evaluation.
Example-2
Ethyl-3- [N- (tetradecanoyl),N-( y-butyrylamido-1-yl)] -amino-3- [(1R, 2R, 3S) l,2-O-isopropylidene-3-0-methyI-l,4-pentofuranos-4-yl])-propionate(7b):
Sieber amide resin (1.1 gm, 0.68 mmol) was placed in solid phase reaction vessel and treated twice with 20% piperidine-DMF (10 mL each) for 5 min and 25 min at room temperature

under N2 agitation. After this resin was successively washed with DMF (3x2 min), iPrOH (1x2 min) and DMF (3x2 min). Fmoc-P- alanine was then added to the resin followed by addition of DIPEA (0.67 mL, 4.09 mmol), TBTu (.66 gm, 2.05 mmol)) and HOBt (0.31 gm, 2.05 mmol) in DMF (10 mL) with N2 agitation for 3 hrs at room temperature. Resin was washed with DMF (3 x 2min) and again treated with Fmoc-y- aminobutyric acid in the presence of DIC (0.32 mL, 2.04 mmol) and HOBT (0.31 gm, 2.05 mmol) for 3 hrs. The resin was washed with DMF (3x2 min), MeOH (3-x 2 min) and DCM (3x2 min). The completion of reaction was monitored by Kaiser test, y- aminobutanoyl-Sieber amide resin (0.05 gm, 0.031 mmol) was placed in the desired reaction block and treated with 20% piperidine-DMF (1 mL) twice for 5 min and 25 min. After this the resin in the well was washed with DMF (3x2 min), iPrOH (1x2 min) and DMF (3 x 2min). This was followed by transfer of the ethyl- [3-(l,2-0-isopropylidene-3-0-methyl)-l,4-pentofuranos-4-yl]-propenoates to the desired well and subsequently treated with DBU (0.10 mL, 0.093 mmol) in DMF/ethanol for 48 hrs. After completion the resins were again washed using the same wash chemfile as described earlier. The resin bound compound in well was then treated with the myristoyl chloride (0.25 ml) in pyridine (2ml) in presence of dimethyl aminopyridine and reaction continued for 24 hrs. The completion of reaction was monitored by negative ninhydrin test. Finally the glycoconjugate was cleaved from the resin using mixture of 2% TFA, 94% dichloromethane and 4%triisopropylsilane. The solvent evaporated and the residue dissolved in t-Butanol/ Water & lyophilized to get the desired compound. The compound wascharacterized using FAB MS: 586 [M+H]+.

Example-3
Ethyl-3- [N- (octanoyl)- N- P-alanylamido-l-yl] -amino-3- [(1R, 2R, 3S) l,2-O-isopropyIidene-3-O-methyl-l, 4-pentofuranos-4-yl])-propanoate (7c):
Sieber amide resin (1.1 gm, 0.68 mmol) was placed in solid phase reaction vessel and treated twice with 20% piperidine-DMF (10 mL each) for 5 min and 25 min at room temperature under N2 agitation. After this resin was successively washed with DMF (3 x 2 min), iPrOH (1x2 min) and DMF (3x2 min). Fmoc-P- alanine was then added to the resin followed by addition of DIPEA (0.67 mL, 4.09 mmol), TBTu (.66 gm, 2.05 mmol)) and HOBt (0.31 gm, 2.05 mmol) in DMF (10 mL) with N2 agitation for 3 hrs at room temperature. Resin was washed with DMF (3 x 2min) and again treated with Fmoc-p-alanine in the presence of DIC (0.32 mL, 2.04 mmol) and HOBT (0.31 gm, 2.05 mmol) for 3 hrs. The resin was washed with DMF (3x2 min). Resin was washed with DMF (3 x 2min) and again treated with Fmoc-P- alanine in the presence of DIC (0.32 mL, 2.04 mmol) and HOBT (0.31 gm, 2.05 mmol) for 3 hrs. The resin was washed with DMF (3 x 2 min), MeOH (3-x 2 min) and DCMF (3x2 min). The completion of reaction was monitored by Kaiser test, p-alanyl acyl-Sieber amide resin (0.05 gm, 0.031 mmol) was placed in the desired reaction block and treated with 20% piperidine-DMF (1 mL) twice for 5 min and 25 min. After this the resin in well was washed with DMF (3x2 min), iPrOH (1x2 min) and DMF (3 x 2min). This was followed by transfer of one of the ethyl- [3-(l,2-O-isopropylidene-3-O-methyl)-l,4-pentofuranos-4-yl]-propenoates to the desired well and subsequently treated with DBU (0.10 mL, 0.093 mmol) in DMF/ethanol for 48 hrs. After completion the resins were again washed using the same wash chemfile as described earlier. The resin in each well was then treated with the octanoyl chloride

(0.25 ml) in pyridine (2ml) in presence of dimethyl aminopyridine and reaction continued for 24 hrs. The completion of reaction was monitored by negative ninhydrin test. Finally the glycoconjugate was cleaved from the resin using mixture of 2% TFA, 94% dichloromethane and 4%triisopropylsilane. The solvent evaporated and the residue dissolved in t-Butanol/ Water & lyophilized to get the desired compound. The compound was characterized using and FAB MS: 488 [M+H]+. Example-4
Ethyl-3- [N- dodecanoyl), N- a-amino butyryl-l-yl] -amino-3- [(1R, 2R, 3S) 1,2-0-isopropylidene-3-O-methyl-l, 4-pentofuranos-4-yl])-propanoate (7d): Sieber amide resin (1.1 gm, 0.68 mmol) was placed in solid phase reaction vessel and treated twice with 20% piperidine-DMF (10 mL each) for 5 min and 25 min at room temperature under N2 agitation. After this resin was successively washed with DMF (3 x 2 min), iPrOH (1x2 min) and DMF (3x2 min). Fmoc-a-amino butyric acid( 6 mmol) was then added to the resin followed by addition of DIPEA (0.67 mL, 4.09 mmol), TBTu (.66 gm, 2.05 mmol)) and HOBt (0.31 gm, 2.05 mmol) in DMF (10 mL) with N2 agitation for 3 hrs at room temperature. Resin was washed with DMF (3 x 2min) and again treated with Fmoc-a-amino butyric acid in the presence of DIC (0.32 mL, 2.04 mmol) and HOBT (0.31 gm, 2.05 mmol) for 3 hrs. The resin was washed with DMF (3x2 min). Resin was washed with DMF (3 x 2min) The resin was washed with DMF (3x2 min), MeOH (3-x 2 min) and DCM (3x2 min). The completion of reaction was monitored by Kaiser test. -a-amino butanoyl acyl-Sieber amide resin (0.05 gm, 0.031 mmol) was placed in the desired reaction block and treated with 20% piperidine-DMF (1 mL) twice for 5 min and 25 min. After this the resin in each well was washed with DMF (3x2 min), iPrOH (1x2

min) and DMF (3 x 2min). This was followed by transfer of the ethyl- [3-( 1,2-0-isopropylidene-3-O-methyl)-l,4-pentofuranos-4-yl]-propenoate to the desired well and subsequently treated with DBU (0.10 mL, 0.093 mmol) in DMF/ethanol for 48 hrs. After completion of the reaction the resins were again washed as above. The resin bound compound in reaction well was then treated with dodecanoyl chloride (0.15 ml) in DMSO (2ml) in presence of DBU (0.1 ml) and reaction continued for 24 hrs. The completion of reaction was monitored by negative ninhydrin test. Finally the glycoconjugate was cleaved from the resin using mixture of 2% TFA, 94% dichloromethane and 4% triisopropylsilane. The solvent evaporated and the residue dissolved in t-Butanol/ Water & lyophilized to get the desired compound. FAB MS: 530 [M+H]+.
E-xample-5
Ethyl-3- [N- (hexadecanoyl), N- P-alanylamido-l-yl] -amino-3- [(1R, 2R, 3S) l,2-O-isopropylidene-3-O-methyl-l, 4-pentofuranos-4-yI])-propionate (7h): Sieber amide resin (1.1 gm, 0.68 mmol) was placed in solid phase reaction vessel and treated twice with 20% piperidine-DMF (10 mL each) for 5 min and 25 min at room temperature under N2 agitation. After this resin was successively washed with DMF (3x2 min), iPrOH (1x2 min) and DMF (3x2 min). Fmoc-P- alanine was then added to the resin followed by addition of DIPEA (0.67 mL, 4.09 mmol), TBTu (.66 gm, 2.05 mmol)) and HOBt (0.31 gm, 2.05 mmol) in DMF (10 mL) with N2 agitation for 3 hrs at room temperature. Resin was washed with DMF (3 x 2min) and again treated with Fmoc-P- alanine in the presence of DIC (0.32 mL, 2.04 mmol) and HOBT (0.31 gm, 2.05 mmol) for 3 hrs. The resin was washed with DMF (3x2 min). Resin was washed with DMF (3 x 2min) and again treated with

Fmoc-p- alanine in the presence of DIC (0.32 mL, 2.04 mmol) and HOBT (0.31 gm, 2.05 mmol) for 3 hrs. The resin was washed with DMF (3x2 min), MeOH (3-x 2 min) and DCM (3x2 min). The completion of reaction was monitored by Kaiser test. P-Alanyl acyl-Sieber amide resin (0.05 gm, 0.031 mmol) was placed in s in the desired reaction block and treated with 20% piperidine-DMF (1 mL) twice for 5 min and 25 min. After this the resin in each well was washed with DMF (3x2 min), iPrOH (1x2 min) and DMF (3 x 2min). This was followed by transfer of the ethyl- [3-(l,2-O-isopropylidene-3-O-methyl)-l,4-pentofuranos-4-yl]-propenoate to the desired well and subsequently treated with DBU (0.10 mL, 0.093 mmol) in DMF/ethanol for 48 hrs. After completion the resins were again washed with DMF (3x2 min), MeOH (3-x 2 min) and DCM (3x2 min). The resin in each well was then treated with the palmitoyl chloride (0.15 ml) in pyridine (2ml) in presence of dimethyl aminopyridine (0.1 ml) and reaction continued for 24 hrs. The completion of reaction was monitored by negative ninhydrin test. Finally the glycoconjugate was cleaved from the resin using mixture of 2% TFA, 94% dichloromethane and 4%triisopropylsilane. The solvent evaporated and the residue dissolved in t-Butanol/ Water & lyophilized to get the desired compound. FAB MS : 572[M+H]+.
ExampIe-6
Ethyl-3- [N- (dodecanoyl), N- (3-aIanylamido-l-yl] -amino-3- [(1R, 2R, 3S) l,2-O-isopropylidene-3-O-benzyI-l, 4-pentofuranos-4-yl])-propanoate (If): Sieber amide resin (1.1 gm, 0.68 mmol) was placed in solid phase reaction vessel and treated twice with 20% piperidine-DMF (10 mL each) for 5 min and 25 min at room temperature under N2 agitation. After this resin was successively washed with DMF (3x2 min), iPrOH (1x2 min) and DMF (3x2 min). Fmoc-P- alanine was then added to the resin followed by

addition of DIPEA (0.67 mL, 4.09 mmol), TBTu (.66 gm, 2.05 mmol)) and HOBt (0.31 gm,
2.05 mmol) in DMF (10 mL) with N2 agitation for 3 hrs at room temperature. Resin was
washed with DMF (3 x 2min) and again treated with Fmoc-P- alanine in the presence of DIC
(0.32 mL, 2.04 mmol) and HOBT (0.31 gm, 2.05 mmol) for 3 hrs. The resin was washed
with DMF (3 x 2 min). Resin was washed with DMF (3 x 2min) and again treated with
Fmoc-P- alanine in the presence of DIC (0.32 mL, 2.04 mmol) and HOBT (0.31 gm, 2.05
mmol) for 3 hrs. The resin was washed with DMF (3x2 min), MeOH (3-x 2 min) and
DCMF (3x2 min). The completion of reaction was monitored by Kaiser test. (3-alanyl acyl-
Sieber amide resin (0.05 gm, 0.031 mmol) was placed in s in the desired reaction block and
treated with 20% piperidine-DMF (1 mL) twice for 5 min and 25 min. After this the resin in
each well was washed with DMF (3x2 min), iPrOH (1x2 min) and DMF (3 x 2min). This
was followed by transfer of one of the ethyl- [3-(l,2-O-isopropylidene-3-O-methyl)-l,4-
pentofuranos-4-yl]-propenoate to the desired well and subsequently treated with DBU (0.10
mL, 0.093 mmol) in DMF/ethanol for 48 hrs. After completion of the reaction the resin
loaded compound were again washed as described earlier. The resin in each well was then
treated with the dodecanoyl chloride (0.15 ml) in pyridine (2ml) in presence of dimethyl
aminopyridine (0.1 ml) and reaction continued for 24 hrs. The completion of reaction was
monitored by negative ninhydrin test. The resin was washed with DMF (3x2 min), iPrOH (1
x 2 min) and DMF (3 x 2mi.n). Finally the glycoconjugate was cleaved from the resin using
mixture of 2% TFA, 94% dichloromethane and 4% triisopropylsilane. The solvent evaporated
and the residue dissolved in t-butanol/ water & lyophilized to get the desired compound. The
compound was characterized using FAB MS: 690 [M+H]+.

Example-7
Ethyl-3- [N- (hexadecanoyl), N- P-alanylamido-l-yl] -amino-3- [(1R, 2R, 3S) l,2-O-isopropylidene-3-O-benzyl-l, 4-pentofuranos-4-yl])-propanoate (7e): Sieber amide resin (1.1 gm, 0.68 mmol) was placed in solid phase reaction vessel and treated twice with 20% piperidine-DMF (10 mL each) for 5 min and 25 min at room temperature under N2 agitation. After this resin was successively washed with DMF (3 x 2 min), iPrOH (1x2 min) and DMF (3x2 min). Fmoc-(3- alanine was then added to the resin followed by addition of DIPEA (0.67 mL, 4.09 mmol), TBTu (.66 gm, 2.05 mmol)) and HOBt (0.31 gm, 2.05 mmol) in DMF (10 mL) with N2 agitation for 3 hrs at room temperature. Resin was washed with DMF (3 x 2min) and again treated with Fmoc-P-alanine in the presence of DIC (0.32 mL, 2.04 mmol) and HOBT (0.31 gm, 2.05 mmol) for 3 hrs. The resin was washed with DMF (3x2 min), MeOH (3-x 2 min) and DCMF (3x2 min). The completion of reaction was monitored by Kaiser test. P-alanyl acyl-Sieber amide resin (0.05 gm, 0.031 mmol) was placed in s in the desired reaction block and treated with 20% piperidine-DMF (1 mL) twice for 5 min and 25 min. After this the resin in each well was washed with DMF (3x2 min), iPrOH (1x2 min) and DMF (3 x 2min). This was followed by transfer of the ethyl- [3-(l,2-O-isopropylidene-3-O-benzyl)-l,4-pentofuranos-4-yl]-propenoate to the desired well and subsequently treated with DBU (0.10 mL, 0.093 mmol) in DMF/ethanol for 48 hrs. After completion the resins were again washed as described earlier. The resin in reaction well was then treated with the hexadecanoyl chloride (0.15 ml) in pyridine (2ml) in presence of dimethyl aminopyridine (0.1 ml) and reaction continued for 24 hrs. The completion of reaction was monitored by negative ninhydrin test. Finally the glycoconjugate was cleaved from the resin using

mixture of 2% TFA, 94% dichloromethane and 4%triisopropylsilane. The solvent evaporated and the residue dissolved in t-butanol/ water & lyophilized to get the desired compound. The compound was characterized using FAB MS : 732[M+H]+.
Example-8
Ethyl-3- [N- (hexadecanoyl), N- p-alanylamido-l-yl] -amino-3- [(1R, 2R, 3S) l,2-O-isopropylidene-3-O-methyl-l, 4-pentofuranos-4-yl])-propanoate (7h): Sieber amide resin (1.1 gm, 0.68 mmol) was placed in solid phase reaction vessel and treated twice with 20% piperidine-DMF (10 mL each) for 5 min and 25 min at room temperature under N2 agitation. After this resin was successively washed with DMF (3x2 min), iPrOH (1x2 min) and DMF (3x2 min). Fmoc-p- alanine was then added to the resin followed by addition of DIPEA (0.67 mL, 4.09 mmol), TBTu (.66 gm, 2.05 mmol)) and HOBt (0.31 gm,
2.05 mmol) in DMF (10 mL) with N2 agitation for 3 hrs at room temperature. Resin was
washed with DMF (3 x 2min) and again treated with Fmoc-p- alanine in the presence of DIC
(0.32 mL, 2.04 mmol) and HOBT (0.31 gm, 2.05 mmol) for 3 hrs. The resin was washed
with DMF (3x2 min), MeOH (3-x 2 min) and DCMF (3x2 min). The completion of
reaction was monitored by Kaiser test. P-alanyl acyl-Sieber amide resin (0.05 gm, 0.031
mmol) was placed in s in the desired reaction block and treated with 20% piperidine-DMF (1
mL) twice for 5 min and 25 min. After this the resin in each well was washed with DMF (3 x
2 min), iPrOH (1x2 min) and DMF (3 x 2min). This was followed by transfer of the ethyl-
[3-(l,2-0-isopropylidene-3-0-benzyl)-l,4-pentofuranos-4-yl]-propenoate to the desired well
and subsequently treated with DBU (0.10 mL, 0.093 mmol) in DMF/ethanol for 48 hrs. After
completion the resins were again washed as described earlier. The resin in reaction well was
then treated with the hexadecanoyl chloride (0.15 ml) in pyridine (2ml) in presence of

dimethyl aminopyridine (0.1 ml) and reaction continued for 24 hrs. The completion of
reaction was monitored by negative ninhydrin test. Finally the glycoconjugate was cleaved
from the resin using mixture of 2% TFA. 94% dichloromethane and 4%triisopropylsilane.
The solvent evaporated and the residue dissolved in t-butanol/ water & lyophilized to get the
desired compound. The compound was characterized using FAB MS : 657[M+H]+.
Example- 9
The library of synthetic compounds was tested against commonest fungal pathogens Candid albicans and Cryptococcus neoformans by microbroth dilution technique (Iwata, K: Bossche, H.V.; In vitro and in vivo Evaluation of Antifungal Agents, Elesevier Science: Amsterdam, 1986, pp 31-34), in Sabouraud's dextrose broth at 28 °C. Stock solutions of the compounds were made in 10% DMSO (dimethyl sulfoxide) and the maximum concentration tested was 100 u.g/ml. The colony forming units (CFU) of both the organisms was maintained at lxlO4/ml in the test broth. The minimal inhibitory concentrations (MICs) were noted based upon absorbance value on an ELISA( Enzyme linked immunosorbent assay) reader at 492 nm after 24 hour (C.albicans) and 48 hour (C neoformans). The antifungal activity of some of the compounds from the library has been summarised in Table 1. Table 1: In vitro antifungal activity of compounds Table-1

Compound No. MICC.albocans Hg/ml C. neoformans
7a 100 100
7b >100 100
7c 25 50
7d 25 50
7f 25 100
7g 25 25
7h 100 100

Example 10
1. For MABA assay M. tuberculosis H37Ra was used as a suitable surrogate for the virulent H37 Rv strain. 7-8 days old cultures were diluted in liquid medium to provide an optimal density of 0.02 at 550 nm spectrophotometrically which gave colour change of Alamar blue 'oxidation reduction' dye (blue to pink).
A compound with MIC Table-2: In vitro antitubercular activity

Compounds MIC(ng /ml)
7b 12.5
7d 12.5
7g 12.5
Rifamycin 0.005
Isoniazid 0.75

Example- 11
The activity of compounds was tested against bioluminiscent Mycobacterium aurum expressing_firefly luciferase (D.K.Deb; K.K.Srivastava; Ranjana Srivastava; and B.S. Srivastava Biochem. Biophys. Res. Comm. 279, 457-61, 2000). The cells were grown to an optical density of 0.03 at 600 nm. Two fold dilutions of compounds were prepared and added to IOOJJI culture (Agoo = 0.03) in microtitre plate. The plate was incubated at 37 °C for 6 hr and bioluminiscence was measured for each well. Two controls (with no drug) and two standard drugs (rifampicin and sparfloxacin) were also included. For measurement of bioluminescence IOOJJ.1 of culture was mixed with 250 \xl of sodium citrate buffer (0.1M, pH 5.0) in the tube and was placed in the luminometer. (Lumat LB 9507- EG & G Berthhold) lOOu.1 of 1 mM luciferase substrate was infected and luminescence was measured as relative light units (RLU) for 10s. The antitubercular activities of the compounds are given in Table-3 and Table-4. Table-3: In vitro antitubercular activity
Treatment Concentration ((^g /ml) RLU
_____ . 1.3 x io6
Rifampicin 2.0 4xlO3
Sparfloxacin 0.5 1.1 x 103
7e 12.5 2xlO4
7f 12.5 1.5 xlO4
7g 12.5 3.3 x 104
7h 12.5 3.1 x 104

TabIe-4:1 n vitro antitubercular activity
Compound/drug MIC (jig/ml)
7b 12l
7c 12.5
7j 12.5
Rifamycin 0.005
Isoniazid 0.75
Table-S: In vitro antitubercular activity

Compound no MIC (ng/ml)
7a 25
7d 12.5
7f 25
7h 50
7i 12.5
Example-12
In macrophage method of screening the combinatorial library the macrophage cell line J744A/ has been cultured in DMEM (Dulbecos modified Eagles medium) medium with 10% FCS at 37 ° C in the presence of 5% CO2. The cultures were infected with 10 m.o.i. of M aurum (grown in sautan medium). After 5hr extracellular bacilli were washed out and the adherent macrophage cells containing mycobacterium were replenished with fresh medium. The drugs were added in different concentration and the infected cultures were reincubated
-25

for a period ranging from 24 to 48 h. After this period the infected macrophages were lysed and plated on nutrient agar tween plates for colony forming units. The data were compared with the untreated samples. In vitro antitubercular activity is given in Table-5

We claim
1. A combinatorial library of 3-substituted amino-3-glycosylated propanoates useful as
antifungal and antibacterial agents of the general formula 7 (figure-1)
(Formula Removed)
wherein R is the alkyl chain consisting of 1-4 carbon alkyl, aralkyl groups or H, R1 is alkyl group or H; R2 is 1-4 carbon branched or unbranched alkyl groups; R3 is branched or unbranched alkyl, heterocycloalkyl or cyclogroup or H; n ranges from 0 to 4, R" is 1 to 4 carbon branched or unbranched alkyl or acyl group, cycloalkyl or heterocycloalkyl groups, aroyl group where aromatic ring is substituted or unsubstituted.
2. A combinatorial library of 3-substituted amino-3-glycosylated propanoates
propanoates useful as antifungal and antibacterial agents as claimed in claim 1, wherein the
representative compounds are as follows
Ethyl-3- [N- (tetradecanoyl), N- -butyralamido- 1 -yl] -amino-3- 1(1R, 2R, 3S) 1,2- 0-isopropylidene-3-0-methyl- 1, 4-pentofuranos-4-ylp-propanoate,
Ethyl-3- [N- (octanoyl), N- p-alanylamido-1 -yll -amino-3- RIR 2R, 38) 1, 2-0- sopropylidene-3-0-methy 1-1, 4-pentofuranos-4-y lD-propanoate,
Ethyl-3- [N- (dodecanoyl), N-(a-aminobutyralmido)-l-yl] -amino-3- RJR, 2R, 35) 1,2-0-lsopropylidene-3-0-methyl-l, 4-pentofuranos-4-ylp-propanoate,
Ethyl-3-[N- (hexadecanoyl), N- P-alanylamido-1-yl] -amino-3- RJR, 2R, 38) 1, 2-0- isopropylidene-3-0-benzy 1 -1,4-pentofuranos-4-y 1D-propanoate,
Ethyl-3-[N- (dodecanoyl), N-p-alanylamido-1-yl] -amino-3- [(IR, 2R, 35) 1,2-0- isopropylidene-3-0-benzy 1 -1, 4-pentofuranos-4-ylp-propanoate,
Ethyl-3-[N- (hexadecanoyl),N-p-alanylamido-l-yl] -amino-3- [(IR, 2R, 35) 1,2-0-isopropyl idene-3-0-benzy 1-1, 4-pentofuranos-4-yl])-propanoate,
Ethyl-3-[N-(hexadecanoyl),N-p-alanylamido-l-yl] -amino-3- [(IR, 2R, 3S) 1,2-0-isopropylidene-3-0-methyl-l, 4-pentofuranos-4-yl])-propanoate,
Ethyl-3-[N-(tetradecyl),N-p-alanylamido-l-yl]-amino-3-[(IR,2R,3S)1,2-0-isopropylidene-3-0-methyl-l,4-pentofuranos-4-yl])-propanoate,
Ethyl-3-[N-(dodecanoyl),N-p-alanylamido-l-yl]-amino-3-[(1R,2R,3S)l,2-0-isopropylidene-3-0-methy 1-1, 4-pento furanos-4-ylp-propanoate,
Ethyl-3-[N-(benzoyl),N-p-alanylamido-l-yl]-amino-3-[(IR,2R,3S)l,2-0-isopropylidene-3-0-methy 1-1, 4-pentofuranos-4-yl])-propanoate,
Ethyl-3-[N-(benzoyl),N-p-alanylamido-l-yl]-amino-3-[(IR,2R,3S)1,2-0-isopropylidene-3-0-benzyl-l, 4-pentofuranos-4-ylp-propanoate,
Ethyl-3-[N- (benzyl),N-p-alanylamido-l-yl] -amino-3- [(IR, 2R, 3S) 1,2-0- isopropyl idene-3 -0-methyl-1, 4-pentofuranos-4-yll)-propanoate,
Ethyl-3-[N-(4-cyanobenzyl),N-p-alanylamido-1 -yl]-amino-3-[(IR,2R,3S) 1,2-0-isopropylidene-3-0-methy 1-1, 4-pentofuranos-4-yl])-propanoat,
Ethyl-3-[N-(dodecanoyl),N-(hexamido-6-yl)]-amino-3-[(IR,2R,3S) 1,2-0-isopropylidene-3-0-methy 1-1, 4-pentofuranos-4-y 1 l)-propanoate,
Ethyl-3-[N-(octanoyl),N-(hexamido-6-yl)-amino-3- [(IR,2R, 3S) 1,2-0- isopropylidene-
3-0-methy 1 -1, 4-pentofuranos-4-y 1])-propanoate,
Ethyl-3-[N-(benzoyl),N-(phenylalanylamido-2-yl)]-amino-3-[(IR,2R,3S) 1,2-0- isopropylidene-
3-0-methyl- I, 4-pentofuranos-4-yl])-propanoate,
Ethyl-3- [N- (benzyl), N-(phenylalanylamido-2-yl)] -amino-3- [(IR,2R,3S) 1, 2-0- isopropylidene-
3-0-methyl- I, 4-pentofuranos-4-yl])-propanoate,
Ethyl-3-[N- (benzyl), N-(phenylalanylamido-2-yl)]-amino-3-[(IR,2R,3S)l,2-0- isopropy lidene-
3-0-benzyl-l, 4-pentofuranos-4-yl])-propanoate,
Ethyl-3- [N- (4-cyanobenzyl), N-(phenylalanylamido-2-yl)l -amino-3- [(1R, 2R. 3S) 1,2-0-
isopropylidene-3-0-methy 1-1, 4-pentofuranos-4-yl])-propanoate,
Ethyl-3- [N- (benzyl), N-(propanamido-2-yl)] -amino-3- [{IR, 2R, 3S) 1,2-0- isopropylidene-
3-0-methyl-l, 4-pentofuranos-4-yl])-propanoate,
Ethyl-3- [N- (benzoyl), N-(propanamido-2-yl)] -amino-3-[(IR,2R,3S) 1,2-0- isopropylidene-
3-0-methyl-l, 4-pentofuranos-4-yl])-propanoate,
Ethyl-3- [N-(dodecanoyl),N-(propanamido-2-yl)] -amino-3- [(IR, 2R 3S) 1,2-0- isopropylidene-
3-0-methyl-l, 4-pentofuranos-4-yl])-propanoate,
Ethyl-3-[N-(tetrdecanoyl),N-(propanamido-2-yl)]-amino-3- [(IR, 2R, 3S)l,2-0- isopropylidene-
3-0-methyl- I, 4-pentofuranos-4-yl])-propanoate,
Ethyl-3 - [N- (pentaEthyl-3- [N- (tetrdecanoyl), N-(propanamido-2-yl)] -am i no-3 - [(1R,
2R, 3S) l,2-0-isopropylidene-3-0-methyl-l, 4-pentofuranos-4-ylD-propanoate decanoyl), N-
(propanamido-2-y1)] -amino-3-[(7R, 2R,35') l,2-0-isopropylidene-3-0-methyl- I, 4-
pentofuranos-4-yl])-propanoate.
3. A combinatorial library of 3-substituted amino-3-glycosylated propanoates useful as antifungal and antibacterial agents substantially as herein described with reference to the examples and drawing accompanying the specifications.

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Patent Number

242121

Indian Patent Application Number

716/DEL/2004

PG Journal Number

34/2010

Publication Date

20-Aug-2010

Grant Date

12-Aug-2010

Date of Filing

15-Apr-2004

Name of Patentee

COUNCIL OF SCIENTIFIC AND INDUSTRIAL RESEARCH

Applicant Address

Rafi Marg, New Delhi.

Inventors:

#	Inventor's Name	Inventor's Address
1	RAMA PATI TRIPATHI	CENTRAL DRUG RESEARCH INSTITUTE, CHATTAR MANZIL PALACE, LUCKNOW-226 001, (U.P.), INDIA.
2	BIJOY KUNDU	CENTRAL DRUG RESEARCH INSTITUTE, CHATTAR MANZIL PALACE, LUCKNOW-226 001, (U.P.), INDIA.
3	PRAVEEN KUMAR SHUKLA	CENTRAL DRUG RESEARCH INSTITUTE, CHATTAR MANZIL PALACE, LUCKNOW-226 001, (U.P.), INDIA.
4	SUDHIR SINHA	CENTRAL DRUG RESEARCH INSTITUTE, CHATTAR MANZIL PALACE, LUCKNOW-226 001, (U.P.), INDIA.
5	RANJANA SRIVASTAVA	CENTRAL DRUG RESEARCH INSTITUTE, CHATTAR MANZIL PALACE, LUCKNOW-226 001, (U.P.), INDIA.
6	KISHORE KUMAR SRIVASTAVA	CENTRAL DRUG RESEARCH INSTITUTE, CHATTAR MANZIL PALACE, LUCKNOW-226 001, (U.P.), INDIA.
7	VINITA CHATURVEDI	CENTRAL DRUG RESEARCH INSTITUTE, CHATTAR MANZIL PALACE, LUCKNOW-226 001, (U.P.), INDIA.
8	ANIL SRIVASTAVA	CENTRAL DRUG RESEARCH INSTITUTE, CHATTAR MANZIL PALACE, LUCKNOW-226 001, (U.P.), INDIA.
9	BRAHM SHANKAR SRIVASTAVA	CENTRAL DRUG RESEARCH INSTITUTE, CHATTAR MANZIL PALACE, LUCKNOW-226 001, (U.P.), INDIA.

PCT International Classification Number

A61K 38/00

PCT International Application Number

N/A

PCT International Filing date

PCT Conventions:

#	PCT Application Number	Date of Convention	Priority Country
1			NA