INTRODUCTION:

Momordica Charantia, Linn belonging to the family Cucurbitaceae. It is called as bitter guard in English , it has been recognized in different systems of traditional medication for the treatment of different diseases and ailments of human beings such as Hypoglycemic activity anti-cancer etc.

METHODOLOGY:

Collection of plant material:

Roots were collected during the month of April 2009 from village Nalwar of Gulbarga District (Karnataka) and was identified and authenticated by Department of Botany, Gulbarga University Gulbarga, a voucher (#172) of specimen was submitted to NGSMIPS, Derlakatte.

Methods extraction

The dried stem bark was powdered (1 1/2 kg) and subjected to soxhlet extraction with increasing polarity of the solvent i.e pet. ether, chloroform, methanol and water. The residues obtained from different solvents were kept in desiccators and the phytochemical investigation was carried out with the extracts obtained from all the solvent.

Quantity of residues obtained from each solvent

1. Pet. Ether 6g

2. Chloroform 12 g ( alkaloids +ve)

3. Methanol 20g ( Flavanoids +ve)

Isolation of Momordica charantia Linn roots

The methanolic extract (25 g) was triturated in mortar with 10ml methanol adsorbed onto silica gel (20 g). After evaporation of the solvent it was loaded onto a silica gel column (150 g) prepared in petroleum ether (60-80˚C). The column was eluted first with petroleum ether (60-80˚C), petroleum ether (60-80˚C): benzene graded mixtures (95:5, 90:10, 80:20, 70:30, 60:40 and 50:50, 60:40,70:30, 80:20, 90:10,100% v/v), then with benzene followed by graded mixtures of benzene: chloroform (95:5, 90:10, 80:20 70:30, 60:40 and 50:50, 60:40,70:30, 80:20, 90:10, 100% v/v), chloroform and finally chloroform: methanol (95:5, 90:10, 80:20, 70:30, 60:40 and (50:50) 60:40,70:30, 80:20, 90:10,100% v/v. The elutions were monitored by TLC (Silica gel-G; visualization by Vanillin - Sulphuric acid reagent heated at 110ºC). The results were given in the

SPECTRAL CHARACTERIZATION

The limitation of the project not reviling the structure coz this are all new molecules and isolated first time, moreover here this are having anti-HIV anti hyperglycemic and highly potent anticancer activity, the another glimpse of the research finding is that we developed the Retro-synthetic methodology on the McReX-1 moiety, which is our spectacular evidence of proof is that the plant is having anti-cancer anti-HIV activity.

Results of elution

} Elution carried out with Pet. Ether 100%, resulting in getting one constituent which gave positive test for Alkaloides. Code [McReX-1]

} Elution carried out with Hexane: Chloroform (80:20) resulting in getting one constituent which gave positive test for Phenanthrene. [McReX-2]

} Elution carried out with Hexane: Chloroform (50:50) resulting in getting one constituent which gave positive test for Phenanthrene. [McReX-3]

} Elution carried out with Chloroform: Methanol (70:30) resulting in getting one constituent which gave positive test for flavonoids and alkaloids [McReX-4]

} Elution carried out with Chloroform: Methanol (80:10) resulting in getting one constituent which gave positive test for alkaloids. [McReX-5]

} Elution carried out with Methanol (100%) resulting in getting one constituent which gave positive test for alkaloids. [McReX-6]

Table of Spectroscopic characterization of constituents.

S.No	S. Code Rf-Value	CHEMICAL NAME
1.	McReX-1 (0.48 )	4-(2-amino-2-(2-(2-hydroxy-3 methylbutyl) octahydropyrrolo [1,2-a]pyrazin-7-yl) ethyl) -2- ethylphenol (retro-synthesized moiety)
2.	McReX-2 (0.53)	9-((2-hydroxy-5-m-tolylpentan-2-yloxy)methyl)-2,10-dimethoxyicosahydro-1H-phenanthro[2,1-f]chromene-1,9-diol
3.	McReX-3 (0.62)	(E)-1-(3-aminophenyl)-7-hydroxy-6-methoxy-3-methyl-7-(1,3,11-trimethoxy-2,4,4,13,14-pentamethyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl)oct-4-en-3-yl acetate.
4.	McReX-4 (0.50)	6-(6-acetyl-2-(11-acetyl-8a-(1-aminoethyl)-4,4,6a,6b,11-pentamethyl-14-oxo1,2,3,4,4a, 5,6,6a,6b,7,8,8a,9,10,11,12,12a,14,14a,14b-icosahydropicen-3-yloxy)-4,5-dihydroxy tetrahydro-2H-pyran-3-yloxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid
5.	McReX-5 (0.55)	14-hydroxy-11-methoxy-10-(2-methoxypropoxy)-4,6a,6b,12,14b-pentamethyl-8a-(methylamino)-4-(1-(2,4,8-trimethyl-2,5,6,7,8,8a-hexahydro-1H-pyrido[3,4-d][1,3]oxazin-6-yloxy)ethyl)octadecahydro-1H-phenanthro[1,2-h]isochromen-3(4H)-one (Activity guided moiety)
6.	McReX-6 (0.16)	6-(17-(4,6-dihydroxy-5-methoxy-2-(methylperoxy)tetrahydro-2H-pyran-3-yloxy)-1,12-dimethoxy-4,5,8,10,12,13,14,17-octahydro-1H-cyclopenta[a]phenanthren-3-yloxy)-2-(methylperoxy)-3,4-dihydro-2H-pyran-3,4,5-triol.

Results and discussion of interpreted moiety.

Summery and discussion of McReX-1

The first molecule of Momordica charantia Linn roots which is isolated from methanolic extract the IR (KBr) spectrum of the compound exhibited the characteristic absorption band in the range of 3441 cm^-1indicates the broad band of hydrogen bonded -OH -CNH groups stretching functionalities of the bioactive molecule, the presence of characteristic stretching peak at 2955 cm^-1 to 2917 cm^-12849 cm^-1 endowments the evidence of –CH₂ and –CH₃, the spectrum further proves that the expected peak at 1635 cm^-1 indicates the stretching of strong R-NH₂group in plane bending of secondary amines. Hence the moieties are attached with the aromatic amide structure at 1472 cm^-1, 1462cm^-1 1378cm^-1. There is a strong indication of RNH2 in plane bending at 1611 cm^-1. The stretching at 1498 cm^-1, 1463 cm^-1 1367cm^-1, and 1329 cm^-1indicates the presence of Ar C-C stretching and Ar-N, this data are supportive to the presence of derivatives of amino group. These indications represent that a molecule is aromatic amide. The H¹NMR of this The H¹NMR spectrum of the compound exhibits characteristics protons signals with different connection in parts per melions (ppm) at 400 MHz in DMSO (δ 2.50) as follows the doublets of peak δ 0.82 to δ 0.88 indicates the presence of -C,-C-R groups of 1- β-CH₃ proton(s), further more the signals at δ 2.56 δ2.69 indicts the presence of -C,-N,-N (C)C,-O-C groups of 1-α, β-CH₂pyrrolidine CH, CH₂ proton(s), the signal at δ 3.34 indicates the presence of C- OH aromatic C-OH proton(s) ,the indication of Sharpe signals at δ 7.63 is due to the presence of -CC groups of 1-benzene of aromatic proton(s), the evidence of presence of –NH₂proton(s) of amide group. HRESI- High-resolution negative ion electrospray mass spectrometry measured and calculated for C₂₂H₃₇N₃O₂ [M _ H]^- m/z 375 (12 %) molecular ion peak and fragmentation peak at m/z 324 (72%) due to release of –C₄H₆O^-+, m/z 309 (100%) intensity gives base peak due to flavonoides structure (C₁₈H₃₅N₃O^-).further release and ionization of C₁₈H₃₅N₃O^- gives on reduction of – C₄H₈O gives fragmented peak on m/z 255 (25%). The evidence of presence of C peaks supportive with C¹³ NMR which are interpreted by comparing the results of IR,NMR, and Mass, it was further proved with the TLC analysis which gave a single spot in solvent system of C₆H₆: Chloroform 5:2 the Rf-0.48 , Quantity of the molecule after analysis is 1g, mp is 250⁰C. The earmark excellencies of above scientific evidence laurels that it is chemically designated [4-(2-amino-2-(2-(2-hydroxy-3-methylbutyl) octahydropyrrolo [1,2-a]pyrazin-7-yl) ethyl)-2- ethylphenol] as respective code is McReX-1, commonly it is a derivatives of protiens, it gave positive test for alkaloids . The interpretation, of next bioactive constituent excels further preceding in our next pearl set tiny molecule, laurels the finding, as follows

Summery and discussion of McReX-2

The third molecule of Momordica charantia Linn roots which is isolated from methanolic extract the IR (KBr) spectrum of the compound exhibited the characteristic absorption band in the range of 3386 cm^-1indicates the broad band of hydrogen bonded -OH groups stretching functionalities of the bioactive molecule, the presence of characteristic stretching peak at 2954 cm^-1 to 2924 cm^-12853 cm^-1 endowments the evidence of –CH₂ and –CH₃, the spectrum further proves that the expected peak at 1635 cm^-1indicates the stretching of-C=C- ,-OCH₃ group in alkene. Hence the moieties are attached with the aromatic amide structure as expected. There is a strong indication at 1401 cm^-1,1405 cm^-1 1378cm^-1Ar C-C bending and rocking. Stretching represent that presence of steroid molecule with aromatic alkenes . The H¹NMR of this compound indicate the proton signals at δ 0.74 to δ 0.96 indicates the presence of -C,-C-R groups of 1- β-CH₃ proton(s), further more the signals at δ 1.07 to δ1.93 indicts the presence of -C,-C(=O)N,-NC(=O)-C groups of 1-α, β-CH₂ cyclohexane proton(s), the signal at δ 2.00 to δ 2.77 indicates the presence of -O-C=C,,-O,-C=O,-C(=O)N,1,2-α β-CH from cyclohexene and methane, OH proton(s), the characteristic signals at δ 3.01 to δ 3.96 indicates the presence of -O,-C=O, -O-C, CH₃ OH proton(s), the characteristic signals at δ 4.02 to δ 4.38 indicates the presence of -O groups of 1-α β-cyclohexadiene CH proton(s), the indication of Sharpe signals at δ 7.63 is due to the presence of -CC groups of 1-benzene of aromatic proton(s), the evidence of presence of –NH₂proton(s) of amide group siglat is observed at δ 8.05. Spectrophotometric analysis of molecule by HRESI- High-resolution negative ion electrospray mass spectrometry measured and calculated for C₃₆H₅₆O₇ [M _ H]^- m/z 600 (14 %) molecular ion peak and fragmentation peak at m/z (72%) due to release of –C₃H₅O^-+, m/z 402 (12%) due to release of –C₁₀H₁₃^-+, m/z 402 (12%) due to release of –C₃H₆O₂^-+, m/z 325(100%) intensity gives base peak due to steroid structure (C₂₁H₂₅O₃^-).further release and ionization of C₂₁H₂₅O₃due to reduction of – H₂O gives fragmented peak on m/z 311 (75%). further release of –C₂H₈ plus H₂O gives fragmented peak on m/z 278. The evidence of presence of C peaks supportive with C¹³ NMR which are interpreted by comparing the results of IR,NMR, and Mass, it was further proved with the TLC analysis which gave a single spot in solvent system of Chloroform: EtOAc 6:4 the Rf-0.53 , Quantity of the molecule after analysis is 3g, mp is 391⁰C. The earmark excellencies of above scientific evidence laurels that it is chemically designated [9-((2-hydroxy-5-m-tolylpentan-2-yloxy)methyl)-2,10-dimethoxyicosahydro-1H-phenanthro[2,1-f]chromene-1,9-diol] as respective code is McReX-2 , commonly it is a derivatives of flavones, it gave positive test for steroids. The interpretation, of next bioactive constituent excels further preceding in our next pearl set tiny molecule, laurels the finding, as follows.

Summery and discussion of McReX-3

The third molecule of Momordica charantia Linn roots which is isolated from methanolic extract the IR (KBr) spectrum of the compound exhibited the characteristic absorption band in the range of 3405 cm^-1indicates the broad band of hydrogen bonded -OH -CNH groups stretching functionalities of the bioactive molecule, the presence of characteristic stretching peak at 2923 cm^-1 to 2852 cm^-1 endowments the evidence of –CH₂ and –CH₃, the spectrum further proves that the expected peak at 1735 cm^-11708 cm^-1 indicates the stretching a strong C=O stretching of ester groups of polycyclic aromatic hydrocarbons (PAH). Hence the moieties are attached with the aromatic amide structure as expected. There is a strong indication of RNH2 in plane bending at 1611 cm^-1. The stretching at 1498 cm^-1, 1463 cm^-1 1367cm^-1, and 1329 cm^-1indicates the presence of Ar C-C stretching and Ar-N flavones with indication at 1269 cm^-11239 cm^-1 shows Ar-C-O stretching represent that presence of flavon molecule with aromatic amide. The H¹NMR of this compound indicate the proton signals at δ 0.74 to δ 0.96 indicates the presence of -C,-C-R groups of 1- β-CH₃ proton(s), further more the signals at δ 1.07 to δ1.93 indicts the presence of -C,-C(=O)N,-NC(=O)-C groups of 1-α, β-CH₂ cyclohexane proton(s), the signal at δ 2.00 to δ 2.77 indicates the presence of -O-C=C,,-O,-C=O,-C(=O)N,1,2-α β-CH from cyclohexene and methane, OH proton(s), the characteristic signals at δ 3.01 to δ 3.96 indicates the presence of -O,-C=O, -O-C, CH₃ OH proton(s), the characteristic signals at δ 4.02 to δ 4.38 indicates the presence of -O groups of 1-α β-cyclohexadiene CH proton(s), the indication of Sharpe signals at δ 7.63 is due to the presence of -CC groups of 1-benzene of aromatic proton(s), the evidence of presence of –NH₂proton(s) of amide group siglat is observed at δ 8.05. Spectrophotometric analysis of molecule by HRESI- High-resolution negative ion electrospray mass spectrometry measured and calculated for C₄₃H₆₄NO₇ [M _ H]^- m/z 710 (19 %) molecular ion peak and fragmentation peak due to release of –C₄H₅N^-+, m/z 642 (30%), due to the release of –H₂O^-+, m/z 625 (20%) due to release of –C₈H₁₁O₂^-+, m/z 485 (25%) further release of –C₅H₉OH indicates base peak of (100%) at m/z 366 due to parent phenanthreen structure .further release and ionization of C₂₃H₄₂O₃due to reduction of – C₅H₁₁ gives fragmented peak on m/z 295 (39%). further release of –C₃H₆ gives fragmented peak on m/z 256 The evidence of .presence of C peaks supportive with C¹³ NMR which are interpreted by comparing the results of IR,NMR, and Mass, it was further proved with the TLC analysis which gave a single spot in solvent system of Chloroform: methanol 6:4 the Rf-0.62 , Quantity of the molecule after analysis is 3g, mp is 405⁰C. The earmark excellencies of above scientific evidence laurels that it is chemically designated [(E)-1-(3-aminophenyl)-7-hydroxy-6-methoxy-3-methyl-7-(1,3,11-trimethoxy-2,4 ,4,13,14-pentamethyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta [a] phenanthren-17-yl)oct-4-en-3-yl acetate] as respective code is McReX-3, commonly it is a derivatives of flavones, it gave positive test for phenols, alkaloid . The interpretation, of next bioactive constituent excels further preceding in our next pearl set tiny molecule, laurels the finding, as follows

Summery and discussion of McReX-4

The third molecule of Momordica charantia Linn roots which is isolated from methanolic extract the IR (KBr) spectrum of the compound exhibited the characteristic absorption band in the range of 3405 cm^-1indicates the broad band of hydrogen bonded -OH -CNH groups stretching functionalities of the bioactive molecule, the presence of characteristic stretching peak at 2923 cm^-1 to 2852 cm^-1 endowments the evidence of –CH₂ and –CH₃, the spectrum further proves that the expected peak at 1735 cm^-11708 cm^-1 indicates the stretching a strong C=O stretching of ester groups of polycyclic aromatic hydrocarbons (PAH). Hence the moieties are attached with the aromatic amide structure as expected. There is a strong indication of RNH2 in plane bending at 1611 cm^-1. The stretching at 1498 cm^-1, 1463 cm^-1 1367cm^-1, and 1329 cm^-1indicates the presence of Ar C-C stretching and Ar-N flavones with indication at 1269 cm^-11239 cm^-1 shows Ar-C-O stretching represent that presence of flavon molecule with aromatic amide. The H¹NMR of this compound indicate the proton signals at δ 0.74 to δ 0.96 indicates the presence of -C,-C-R groups of 1- β-CH₃ proton(s), further more the signals at δ 1.07 to δ1.93 indicts the presence of -C,-C(=O)N,-NC(=O)-C groups of 1-α, β-CH₂ cyclohexane proton(s), the signal at δ 2.00 to δ 2.77 indicates the presence of -O-C=C,,-O,-C=O,-C(=O)N,1,2-α β-CH from cyclohexene and methane, OH proton(s), the characteristic signals at δ 3.01 to δ 3.96 indicates the presence of -O,-C=O, -O-C, CH₃ OH proton(s), the characteristic signals at δ 4.02 to δ 4.38 indicates the presence of -O groups of 1-α β-cyclohexadiene CH proton(s), the indication of Sharpe signals at δ 7.63 is due to the presence of -CC groups of 1-benzene of aromatic proton(s), the evidence of presence of –NH₂proton(s) of amide group siglat is observed at δ 8.05. Spectrophotometric analysis of molecule by HRESI- High-resolution negative ion electro spray mass spectrometry measured and calculated for C₄₄H₆₇NO₁₄ [M _ H]^- m/z 833 (10 %) molecular ion peak and fragmentation peak due to release of –C₂H₃O^-+, m/z 795 (13%), due to the release of –C ₂H₆N^-+, m/z 748 (20%) due to release of –C₂H₆, m/z 716 (19%) further release of –CHO₂ gives m/z 669 (25%) further reduction of –C₅H₈O^2-,m/z 508 (14%) due to release of –C₅H₉O₃gives peak at m/z 399 (15%), the release of -C₃H₆O indicates base peak of (100%) intencity at m/z 325 which indicates due to parent phenanthreen structure .further release and ionization of C₂₃H₃₃Odue to reduction of – CH₃ gives fragmented peak on m/z 311 (73%). further release of –C₄H₆ in the molecule it gives fragmented peak on m/z 264 (13%). The evidence of presence of C peaks supportive with C¹³ NMR which are interpreted by comparing the results of IR,NMR, and Mass, it was further proved with the TLC analysis which gave a single spot in solvent system of EtOAc:methanol:water 4:5:1 the Rf-0.62, Quantity of the molecule after analysis is 3g, mp is 225⁰C. The earmark excellencies of above scientific evidence laurels that it is chemically designated [6-(6-acetyl-2-(11-acetyl-8a-(1-aminoethyl)-4,4,6a,6b,11-pentamethyl-14-oxo 1 ,2, 3,4,4a, 5,6,6a,6b,7,8,8a,9,10,11, 12,12a,14,14a,14b-icosahydropicen-3-yloxy)-4,5-dihydroxytetrahydro-2H-pyran-3-yloxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid (Anti- diabetic activity)] as respective code is McReX-4, commonly it is a derivatives of flavones, it gave positive test for aldehydes and flavonides. The interpretation, of next bioactive constituent excels further preceding in our next pearl set tiny molecule, laurels the finding, as follows

Summery and discussion of McReX-5

The third molecule of Momordica charantia Linn roots which is isolated from methanolic extract the IR (KBr) spectrum of the compound exhibited the characteristic absorption band in the range of 3405 cm^-1indicates the broad band of hydrogen bonded -OH -CNH groups stretching functionalities of the bioactive molecule, the presence of characteristic stretching peak at 2923 cm^-1 to 2852 cm^-1 endowments the evidence of –CH₂ and –CH₃, the spectrum further proves that the expected peak at 1735 cm^-11708 cm^-1 indicates the stretching a strong C=O stretching of ester groups of polycyclic aromatic hydrocarbons (PAH). Hence the moieties are attached with the aromatic amide structure as expected. There is a strong indication of RNH2 in plane bending at 1611 cm^-1. The stretching at 1498 cm^-1, 1463 cm^-1 1367cm^-1, and 1329 cm^-1indicates the presence of Ar C-C stretching and Ar-N flavones with indication at 1269 cm^-11239 cm^-1 shows Ar-C-O stretching represent that presence of flavon molecule with aromatic amide. The H¹NMR of this compound indicate the proton signals at δ 0.74 to δ 0.96 indicates the presence of -C,-C-R groups of 1- β-CH₃ proton(s), further more the signals at δ 1.07 to δ1.93 indicts the presence of -C,-C(=O)N,-NC(=O)-C groups of 1-α, β-CH₂ cyclohexane proton(s), the signal at δ 2.00 to δ 2.77 indicates the presence of -O-C=C,,-O,-C=O,-C(=O)N,1,2-α β-CH from cyclohexene and methane, OH proton(s), the characteristic signals at δ 3.01 to δ 3.96 indicates the presence of -O,-C=O, -O-C, CH₃ OH proton(s), the characteristic signals at δ 4.02 to δ 4.38 indicates the presence of -O groups of 1-α β-cyclohexadiene CH proton(s), the indication of Sharpe signals at δ 7.63 is due to the presence of -CC groups of 1-benzene of aromatic proton(s), the evidence of presence of –NH₂proton(s) of amide group siglat is observed at δ 8.05. Spectrophotometric analysis of molecule by HRESI- High-resolution negative ion electrospray mass spectrometry measured and calculated for C₄₄H₇₅N₃O₈ [M _ H]^- m/z 775 (23 %) molecular ion peak and fragmentation peak due to release of –C₄H₉O^-+, m/z 702 (16%), due to the release of –C H₅N^-+, m/z 671 (40%) due to release of –CH₃O₂H, m/z 624 (12%) further release of –C₅H₉ gives m/z 554 (12%) further reduction of –C₁₀H₁₇N₂O^-,m/z 376 (70%) due to release of –C₃H₈Ogives peak at m/z 323 indicates base peak of (100%) intensity phenanthreen structure, the release of –C₂H₆O₂ at m/z 255 which indicates fragmented peak (39%). The evidence of presence of C peaks supportive with C¹³ NMR which are interpreted by comparing the results of IR,NMR, and Mass, it was further proved with the TLC analysis which gave a single spot in solvent system of EtOAc:methanol:water 2:7:1 the Rf-0.55 , Quantity of the molecule after analysis is 3g, mp is 329⁰C. The earmark excellencies of above scientific evidence laurels that it is chemically designated [14-hydroxy-11-methoxy-10-(2-methoxypropoxy)-4,6a,6b,12,14b-pentamethyl-8a-(methylamino)-4-(1-(2,4,8-trimethyl-2,5,6,7,8,8a-hexahydro-1H-pyrido[3,4-d][1,3]oxazin-6-yloxy)ethyl)octadecahydro-1H-phenanthro[1,2-h]isochromen-3(4H)-one] as resp ective code is McReX-5, commonly it is a derivatives of flavones, it gave positive test for aldehydes and flavonides. The interpretation, of next bioactive constituent excels further preceding in our next pearl set tiny molecule, laurels the finding, as follows

Summery and discussion of McReX-6

The third molecule of Momordica charantia Linn roots which is isolated from methanolic extract the IR (KBr) spectrum of the compound exhibited the characteristic absorption band in the range of 3405 cm^-1indicates the broad band of hydrogen bonded -OH -CNH groups stretching functionalities of the bioactive molecule, the presence of characteristic stretching peak at 2923 cm^-1 to 2852 cm^-1 endowments the evidence of –CH₂ and –CH₃, the spectrum further proves that the expected peak at 1735 cm^-11708 cm^-1 indicates the stretching a strong C=O stretching of ester groups of polycyclic aromatic hydrocarbons (PAH). Hence the moieties are attached with the aromatic amide structure as expected. There is a strong indication of RNH2 in plane bending at 1611 cm^-1. The stretching at 1498 cm^-1, 1463 cm^-1 1367cm^-1, and 1329 cm^-1indicates the presence of Ar C-C stretching and Ar-N flavones with indication at 1269 cm^-11239 cm^-1 shows Ar-C-O stretching represent that presence of flavon molecule with aromatic amide. The H¹NMR of this compound indicate the proton signals at δ 0.74 to δ 0.96 indicates the presence of -C,-C-R groups of 1- β-CH₃ proton(s), further more the signals at δ 1.07 to δ1.93 indicts the presence of -C,-C(=O)N,-NC(=O)-C groups of 1-α, β-CH₂ cyclohexane proton(s), the signal at δ 2.00 to δ 2.77 indicates the presence of -O-C=C,,-O,-C=O,-C(=O)N,1,2-α β-CH from cyclohexene and methane, OH proton(s), the characteristic signals at δ 3.01 to δ 3.96 indicates the presence of -O,-C=O, -O-C, CH₃ OH proton(s), the characteristic signals at δ 4.02 to δ 4.38 indicates the presence of -O groups of 1-α β-cyclohexadiene CH proton(s), the indication of Sharpe signals at δ 7.63 is due to the presence of -CC groups of 1-benzene of aromatic proton(s), the evidence of presence of –NH₂proton(s) of amide group siglat is observed at δ 8.05. Spectrophotometric analysis of molecule by HRESI- High-resolution negative ion electrospray mass spectrometry measured and calculated for C₃₈H₅₃NO₉ [M _ H]^- m/z 667 (5 %) molecular ion peak and fragmentation peak at m/z 634 (3%) due to release of –C₂H₆^-+, m/z 559 (2%) due to release of –C₆H₅O^-+,m/z 517 (30%) due to release of –C₃H₄,m/z 402 (18%)due to release of –C₈H₁₈O⁺,m/z 326 (100%) intensity gives base peak due to flavonoides structure (C₁₅H₁₈O₈^-) .further release and ionization of C₁₅H₁₈O₈^- gives on reduction of -H₂O gives fragmented peak on m/z 311 (36%) and due to release of –C₂H₄O_2, m/z 255 (10%). The evidence of presence of C peaks supportive with C¹³ NMR which are interpreted by comparing the results of IR,NMR, and Mass, it was further proved with the TLC analysis which gave a single spot in solvent system of EtOAc:methanol 7:3 the Rf-0.25 , Quantity of the molecule after analysis is 4g, mp is 329⁰C. The earmark excellencies of above scientific evidence laurels that it is chemically designated [6-(17-(4,6-dihydroxy-5-methoxy-2-(methylperoxy)tetrahydro-2H-pyran-3-ylox y)-1,12-dimethoxy-4,5,8,10,12,13,14,17-octahydro-1H-cyclopenta[a]phenanthren-3-yloxy)-2-(methylperoxy)-3,4-dihydro-2H-pyran-3,4,5-triol] as respective code is CaReX-6, commonly it is a derivatives of triterpenoids, it gave positive test for phenanthren alcohol.

PHARMACOLOGICAL SCREENING:

Induction of diabetes

The animals were starved overnight and then diabetes was induced by a single intravenous injection of a freshly prepared streptozotocin (STZ) solution (50 mg/kg body wt). Streptozotocin was dissolved in 0.1 m freshly prepared citrate buffer solution (pH 4.5). The animals were allowed to drink 5% glucose solution overnight. After 5 days, streptozotocin administration, rats showed diabetes. Control rats were injected with citrate buffer alone. The McReX-5 in aqueous solution was administered orally through gastric catheter at a concentration of 3 mg/kg body weight/rat, once a day for 7 days.

Experimental design

The animals were divided into four groups for the analysis of biochemical parameter.

Each group has 6 animals.

Group I : Normal control rats.

Group II : Diabetic control rats.

Group III : Diabetic rats treated with McReX-4, 3 mg/kg body wt orally.

Group IV : Diabetic treated with standard drug Glibencalimide.

Table No-2 Percentage reduction of blood glucose level in STZ induced diabetic albino rats

Groups	Blood glucose level (Before treatment)	Blood glucose level (After treatment)	Percentage reduction in blood glucose level
Control	268.67 ± 5.13	267.67 ± 6.89	0.37 %
McReX-5 3mg/kg	264.17 ± 9.60	155.33 ± 2.33	80.18 %
Glibencalimide	263.34 ± 4.33	199.33 ± 1.67	70.91 %

As shown in Table 2, the moiety McReX-5 significantly reduced the blood glucose level in STZ induced diabetic rats. The anti-hyperglycemic activity of the molecule observed was better compared to the dose of standard. In diabetic albino rats, maximum percentage reduction was found to be 80.18% and 70.91%, respectively for the standard drug. it have been observed that maintenance of blood glucose level in normal and streptozotocin induced diabetic rats very better as compared with standard.

CONCLUSION:

For centuries, indigenous drugs either alone or in combination have been advocated in the traditional system of medicine for the treatment of various ailments. India is rich in plant wealth and has excellent base of utility of plants with ayurvedic science. The Object of the present research was merely depends on the isolation and establishment of structure of the phytoconstituents which are responsible for the antidiabetic activity and retrosynthatic development of the moites the further work on this plant is gating progress in ouer research laboratory of NGSMIPES mangalore, VIP sciences Rajamundry AIKTC School Of Pharmacy New Parnvel Mumbai.

ACKNOWLEDGEMENT:

The author thanks to the management of AIKTC School of pharmacy and especially I feel immense pleasure to thanks Mr.Abdul Razzaqh Hanutgi, Mrs. Rajni Shattigaru, and Mr. Abusufian Shaikh for providing the platform to carry out the entitled work.

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Received on 20.02.2014 Modified on 11.03.2014

Asian J. Research Chem. 7(3): March 2014; Page 256-261