INTRODUCTION:

The plant Momordica charantia Linn (Family: Cucurbitaceae) is a well known plant and widely distributed and cultivated in many parts of India. It is known as bitter gourds in English, Pavakay in Tamil, Karela in Hindi and Bengali, Karke in Marathi, and Kaippa or Kaippa-valli in Malayalam [1]. The fruit of the plant is widely used as vegetable as well as it is used in Ayurvedic and Unani system of medicines for the treatment of many diseases. The fruits and leaves of M. charantia Linn are useful in piles, leprosy, jaundice, diabetes, snake-bite and it is found to have vermifuge and antioxidant properties [2]. The earlier reports showed that the plant also has anti- malarial, anti-plasmodial properties [3, 4] and insecticidal activity against mustered sawfly [5]. The present study was carried out to investigate the anxiolytic, antidepressant and anti-inflammatory potential of methanol extract of dried leaves of Momordica charantia Linn.

GEOGRAPHICAL SOURCE:

Momordica charantia (Bitter melon or Bitter guard) is a flowering vine in the family Cucurbitaceae:

Leaves:

Simple, usually palmately 5-7 lobed, tendrils unbranched or 2 branched. The herbaceous, tendril bearing vine grows to 5 m. It bears simple, alternate leaves 4–12 cm across, with 3–7 deeply separated lobes.

Fruit:

Ovoid, ellipsoid, or spindle shaped, usually ridged or warty, dehiscent irregularly as a 3 valved fleshy capsule or indehiscent. The fruit has a distinct warty looking exterior and an oblong shape. It is hollow in cross-section, with a relatively thin layer of flesh surrounding a central seed cavity filled with large flat seeds and pith. Seeds and pith appear white in unripe fruits, ripening to red; the flesh is crunchy and watery in texture, similar to cucumber, chayote or green bell pepper. The skin is tender and edible. The fully ripe fruit turns orange and mushy. Bitter melon comes in a variety of shapes and sizes. The typical Chinese phenotype is 20–30 cm long, oblong with bluntly tapering ends and pale green in color, with a gently undulating, warty surface. The bitter melon more typical of India has a narrower shape with pointed ends, and a surface covered with jagged, triangular "teeth" and ridges. Coloration is green or white. Between these two extremes is any number of intermediate forms. Some bear miniature fruit of only 6–10 cm in length, which may be served individually as stuffed vegetables. These miniature fruit are popular in Southeast Asia as well as India. In Panama bitter melon is known as Balsamino. The pods are smaller and bright orange when ripe with very sweet red seeds.

Flowers:

Staminate flowers usually solitary on a bracteates scape, hypanthium shallow, calyx 5 lobed, petals 5, usually yellow, distinct, 1-3 with incurved scales at base, stamens usually 3, inserted toward base of hypanthium, filaments distinct, broad, anthers distinct or coherent, 2 of them dithecal, the other monothecal, cells curved or flexuous; pistillate flowers usually solitary on a bracteates scape, hypanthium ovoid to spindle shaped, perianth usually smaller than in staminate flowers, staminodes absent or 3, ovules numerous, horizontal, stigmas 3, 2 lobed. Seeds few to numerous, ovate, usually sculptured. Each plant bears separate yellow male and female flowers. Though it has been claimed that bitter melon’s bitterness comes from quinine, no evidence could be located supporting this claim.

DISTRIBUTION:

The original home of the species is not known, other than that it is a native of the tropics. Bitter melon grows in tropical areas, including parts of the Amazon, east Africa, Asia, and the Caribbean. It is widely grown in India and other parts of the Indian subcontinent, Southeast Asia, China, Africa, and the Caribbean.

CULTIVATION:

It is a genus of annual or perennial climbers found throughout India and is also cultivated upto an altitude of 1500m. It is cultivated during warm season i.e. during April to July by using 2-3 seeds in a pit. The pits are prepared at a distance of half a meter and provided with manures. Only one plant is retained and seedlings are watered once or twice a week. Plants begin to flower 30-35 days after sowing and the fruits are ready for harvesting 15-20 days after flowering. Bitter gourd, also known as balsam pear, is a tropical vegetable widely cultivated in Asia, Africa and South America [5, 6]. M. charantia fruits consists glycosides, saponins, alkaloids, reducing sugars, resins, phenolic constituents, fixed oil and free acids. M. Charantia consists the following chemical constituents those are Alkaloids, charantin, charine, cryptoxanthin, cucurbitins, cucurbitacins, cucurbitanes, cycloartenols, diosgenin, elaeostearic acids, erythrodiol, galacturonic acids, gentisic acid, goyaglycosides, goyasaponins, guanylate cyclase inhibitors, gypsogenin, hydroxytryptamines, karounidiols, lanosterol, lauric acid, linoleic acid, linolenic acid, momorcharasides, momorcharins, momordenol, momordicilin, momordicins, momordicinin, momordicosides, momordin, momordolo, multiflorenol, myristic acid, nerolidol, oleanolic acid, oleic acid, oxalic acid, pentadecans, peptides, petroselinic acid, polypeptides, proteins, ribosome-inactivating proteins, rosmarinic acid, rubixanthin, spinasterol, steroidal glycosides, stigmasta-diols, stigmasterol, taraxerol, trehalose, trypsin inhibitors, uracil, vacine, v-insulin, verbascoside, vicine, zeatin, zeatin riboside, zeaxanthin, zeinoxanthin Amino acids-aspartic acid, serine, glutamic acid, thscinne, alanine, g-amino butyric acid and pipecolic acid, ascorbigen, b-sitosterol-d-glucoside, citrulline, elasterol, flavochrome, lutein, lycopene, pipecolic acid. The fruit pulp has soluble pectin but no free pectic acid. Research has found that the leaves are nutritious sources of calcium, magnesium, potassium, phosphorus and iron; both the edible fruit and the leaves are great sources of the B vitamins[7,8].

Extraction and isolation:

Packing method: - Wet Packing.

Preparation of extract for column : - 25g of ME in 10ml methanol with 20 g silica gel was triturated in mortar, evaporate the solvent from the triturated mass and loaded onto a column contains silica gel 150 g.

Elution by column:

The packed 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, 50:50, 40:60,30:70, 20:80, 10:90,100% v/v), then with benzene followed by graded mixtures of benzene: chloroform (95:5, 90:10, 80:20, 70:30, 60:40, 50:50, 40:60,30:70, 20:80, 10:90,100% v/v), chloroform and finally chloroform: methanol (95:5, 90:10, 80:20, 70:30, 60:40, 50:50, 40:60,30:70, 20:80, 10:90,100% v/v). The elations were monitored by TLC (Silica gel-G; visualization by Vanillin - Sulphuric acid reagent heated at 110ºC). Elution carried out with benzen: Chloroform (50:50) resulting in getting one constituent which gave positive test for flavonoids. [CaReX-1]

Spectral analysis of molecule:

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- ethyl phenol] 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:

McReX-1

¹HNMR (400 MHz DMSO) δ 0.88, (dd, J= 3 Hz, 7 H), δ 2.69 (t, J= 8 Hz, 10 H), δ 3.34 (dd, J= 6 Hz, 10 H), δ 7.63, (d, J= 1.50 Hz, 9 H), ¹³C NMR (400 MHz DMSO) δ 129.07 (C1), 151.26 (C 2) ,134.65 (C 3), 118.62 (C 4), 135.46 (C 5 ) , 121.09 (C 6 ), 23.09 (C 7), 12.29 (C 8) , 41.01 (C 9), 52.53 (C 10), 43.22 (C 11 ), 64.74 (C 12 ), 54.56 (C 13), 34.83 (C 14), 53.83 (C 15 ), 52.49 (C 16), 56.35 (C 17), 57.06 (C 18 ), 70.09 (C 19), 32.55 (C 20), 18.78 (C 21), 18.78 (C 22 ); IR (KBr), 3441 cm^-1 (sh, OH,CNH ), 2955 cm^-1 , 2917 cm^-1 , 2849 cm^-1 (C-C str), 1635 cm^-1 (R-NH₂ str ), 1472 cm^-1 1462 cm^-1 1378 cm^-1 (Ar-C-C , Ar-N bending), 800 to 720 cm^-1(ortho substituted group),

Development of retrosynthetic methodology of molecule:

Physical state: White colure solid

Solubility: Freely soluble in Methanol and DMSO

Fraction collected: 100% pet. ether

Bioactive molecule picked for retro-synthetic development:

Chemical name:

4-(2-amino-2-(2-(2-hydroxy-3methylbutyl) octahydropyrrolo [1,2-a]pyrazin-7-yl) ethyl)-2- ethylphenol

Chemical Formula: C₂₂H₃₇N₃ O₂

Molecular Weight: m/z 375

Melting point:

302ºC It gave positive test for alkaloids,

UV spectroscopy:

In methanol it gave wavelength maximum at 235 nm.

Toxicological:

Do not shows any toxicity coz drug is having food value

MATERIALS AND METHODS:

Methodology adopted for total synthesis by selecting isolated molecules of Momordica charantia roots called McReX-1:

· Alkenes are wonderful because they are readily functionalized

· They are large number of reliable way for generating them

· Bromide groups are selected for functional group conversion

Strategy planed for synthesis

ANALYSIS: Retro-synthesis; the strategy of Disconnection approach to the McReX-1

Synthetic schemes:

Octahydropyrrolo [1,2-a]pyrazine (1M). Take 0.01mol of piperzine (1 g) in a solution of 1, 3-dibromopropane and pinch of zinc dust (2 g, 0.01 g), the mixture was starred for 2 hr. transfer the whole content to RB-flask with solvent (25 ml), reflex for 7 hr at 40⁰C. Remove the content transfer it in to ice cold water, filter on Buckner funnel, and the dry residue is purified by recrystallized with EtOH to provide 3.5 g (85%) of 1M. IR (KBr) 3417 cm^-1 ( str, NH,), 2960 cm^-1 ( str C-C), 1451,1413,1380 cm^-1(ben, roc C-C),1328 cm^-1(str N-H); ¹HNMR (400 MHz DMSO) δ 1.67 (d, J= 4 Hz, 1 H), 2.47 (q, J= 8, 4 Hz, 4 H), 2.63 (d, J= 16 Hz 1 H), 3.09 (q, J= 8, Hz 2 H), 3.97 (s, 6H ); ¹³C NMR (100 MHz DMSO) δ 23.24 (C 7), 23.44(C 8), 40.04 (C 3), 43.06 (C 1),50.26 (C 4), 55.93 (C 8a); HRESIMS calculated for m/z C₇H₁₄N₂ [M+H]⁺ m/z 127.2 (100%), [M-H]^- m/z 121 (5 %) intencity. Colorless crystals (soluble in DMSO); mp 138⁰C, Rf-value 0.9 (hexane: acetone 1:1).

Synthetic reaction schem-2:

1-(hexahydropyrrolo [1,2-a]pyrazin-2(1H)-yl)-3-methylbutan-2-ol (2 M):

Dehydrohalogenations of 1-bromo-3-methylbutane (0.1 mol, 7 g) in KOH (0.1 mol ,2 g) dissolve in (10 ml) of THF gives 3-methyl-1-pentene stirred for 3hr after that hole content is transfer to RB-flax reflex for 1hr at 50⁰C. Remove the content transfer it in to ice cold water, filter on Buckner funnel, and the dry residue is purified by recrystallized with EtOH to provide 6. g (83%) 3-methylbut-1-ene. Whereupon brominating ofmethylbut-1-ene (0.1 mol 3.5g and 1ml Br₂) gives 1,2-dibromo-3-methylbutane 6.9g (86%). Purified the residue by recrystalization with EtOH to provide 6 g (81%) Condensation of 1,2-dibromo-3-methylbutane with 1 M (0.1mol , 7 g and 0.1mol 11 g) in presence of K₂CO₃(3 g) KOH (1 g) solution, starr the content for 4 hr, reflex it for 8 hr at 50⁰C. Remove the content transfer it in to ice cold water, filter on Buckner funnel, and the dry residue is purified by recrystallized with EtOH to provide 9 g (90%) of pure M 2 ¹HNMR (400 MHz DMSO) δ 0.89,0.87 (dd, J= 8 Hz, 9 H), 1.35 (q, J= 8 Hz, 1H), 1.63 (q, J= 4 Hz, 3 H), 1.79 (s, J= 6 Hz 1 H ) 2.32 ( dd, J= 8, 4 Hz, 2 H), 2.41 (s, 1 H ), 2.59 (t, J= 4 Hz, 3 H), 3.67 ( s, 1 H ); ¹³C NMR (100 MHz, DMSO) δ 22.41 ( C 4), 23.66 ( C 1 ), 24.74 ( C 7 ), 25.97 ( C 8 ), 35.76 ( C 3 ), 52.62 ( C 3 ), 52.72 ( C 4 ), 52. 72 ( C 6 ), 56.04 ( C 1), 56. 28 ( C 1 ), 77. 95 ( C 2 ), 78.61 ( C 8a) ; IR (KBr), 3850 cm^-1(s, OH), 3448 cm^-1(str N-H),2951,2838 cm^-1(str, C-C),1474 cm^-1(C-H ben),1271 cm^-1(C-N). HRESIMS calculated for m/z C₁₂H₂₄N₂O [M+H]⁺ m/z 210.3 (10%), [M-H]^- m/z 211 (5 %) intencity. Colorless crystals (soluble in DMSO); mp 180⁰C, Rf-value 0.5 (hexane:acetone 1:1).

Synthetic reaction schem-3 2-amino-2-(2-(2-hydroxy-3-methylbutyl)octahydropyrrolo[1,2-a]pyrazin-7-yl)ethan-1-ylium (M 3) . Dehydrohalogenation of 2-bromoethanamine HBr in KOH solution (0.1 mol 6 g, 0.1 mol 2.8 g ). Heat on a water bath for 4 hr, cool the content and filter it on Buckner funnel, purified by recrystalizatin with EtOH to provide 7 g (78 %) (Part-I ethenamine). Brominations of Part-I (0.1 mol 6.5 g, 1ml Br₂) stirred it for 1 hr and transfer this (Part-II 1-bromoethanamine) in two portions to RB-flask, with (0.1mol 8 g) M 2 in a solution of K₂CO₃,THF and HCl ( 1 g, 30 ml, 2 ml), reflux it for 12 hr at 45⁰C. Remove the content transfer it in to ice cold water, filter on Buckner funnel, and the dry residue is purified by recrystallized with EtOH to provide 9 g (90%) of pure M 3.¹

HNMR (400 MHz DMSO) δ 0.89,0.87 (dd, J= 8 Hz, 9 H), 1.34 (d, J= 8 Hz, 1 H), 1,37 (dd, J= 11 Hz, 2 H), 1.58(t, J= 12 Hz, 1 H), 1.63(dd, J= 4 Hz, 1 H), 1.83 (s, 1 H), 2.14(t, 1 H),2.33 (dd, J= Hz 8, 4 H),2.41 (d 1H), 2.59 (multip, J= 4 Hz, 6 H), 3. 59 (d, J= 4 Hz, 2 H);¹³C NMR (100 MHz DMSO) δ 22.39 (C 4), 23.67 (C 2), 24.05(C), 25.99(C 3), 35.26 (C 8), 39.85 (C 7), 40.05 (C 1), 52.63 (C 6), 52.76 ( C 4), 56.04 ( C 1), 57.04 ( C 1), 77.84 (C 8a), 78.17 ( C 2 ); IR (KBr), 3422 cm^-1( s NH2, sh OH), 2951 cm^-1( s C-C), 1650 cm^-1(ben N-H), 1474 cm^-1,1383 cm^-1(C-H ben in C-C). ). HRESIMS calculated for m/z C₁₄H₂₉N₃O [M+H]⁺ m/z 253.4 (5%), [M-H]^- m/z 254 (30 %) intensity. Colorless crystals (soluble in DMSO); mp 203 ⁰C, Rf-value 0.6 (hexane:acetone 1:1).

Fig-141:- Synthetic reaction schem-4 2-(2-amino-2-(2-(2-hydroxy-3-methylbutyl) octahydropyrrolo [1, 2-a] pyrazin-7-yl) ethyl)-6-ethylphenol (4 M), Condensation of 2-bromo-6-ethylphenol with M 3 (0.1 mol 10 g, 0.1 mol 10g) in a solution of K₂CO₃, 1, 2-dioxan, THF (1 g, 3 ml and 25 ml), stirred it for 6 hrs at rt, transfer the whole content in to RB-flask, reflex for 10 hr at 65⁰C. Remove the content transfer it in to ice cold water, filter on Buckner funnel, and the dry residue, purified by recrystallized with EtOH to provide 6 g (90%) of pure M 4. ¹HNMR (400 MHz DMSO) δ 0.89, (dd, J= 4 Hz, 9 H), 1.16 (t, J= 8 Hz, 3 H),1.33 (d, J= 8 Hz, 2 H), 1.61 (q, J= 8 Hz, 4 H), 1.85 ( s, 1 H), 2.28 ( q, J= 8 Hz, 2 H), 2.32( J= 8 Hz, 2 H), 2.52 (q, J= 8 Hz, 2 H), 2.32 (q J= 8 Hz, 2 H), 2.55(d J= 8 Hz, 2 H), 3.62 ( d, 1H ) 3.63 ( d, 1 H) 3.66 (d, 2 H), 4.13 ( s, 3 H), 6.69 ( dd, J= 4,8 Hz 4 H), 6.96 ( dd, J= 4,8 Hz 4 H );¹³C NMR (100 MHz DMSO) δ 15.87 (C 2), 22.46 (C 4) ,23.65 (C 1), 25.93(C 3), 27.39 (C 8 ) , 35.27 (C 2 ),38.97 (C 7), 39.39 (C 3) ,39.81 (C 4), 40.02 (C 1), 52.75(C 6 ), 56.22 (C 1 ), 67.07 (C 1), 78.15 (C 8a), 78.48 (C 2 ), 78.64 (C 3), 78.80 (C 5), 114.89 (C 4 ), 128. 19 (C 2), 133.75 (C 1), 155.72 (C 6 ); IR (KBr) 3544 cm^-1 (N-H s ), 3505 cm^-1 (sh, OH), 2957 cm^-1 (C-C str), 1585 cm^-1 (C-C str aromatic ring),1614 cm^-1 (-C=C-),1454 cm^-1( C-H bend), 1246 cm^-1(C-N), 943 cm^-1(N-H wag),725 cm^-1 (C-C rok); HRESIMS calculated for m/z C₂₂H₃₇N₃O₂ [M+H]⁺ m/z 376.4 (10%), [M-H]^- m/z 275 (30 %) intensity. White powder (soluble in DMSO); mp 221 ⁰C, Rf-value 0.7 (hexane:acetone 1:1). In the course of the investigation, the glimpse of research finding got its conclusion by reporting the thirteen (13), pure medicinally active secondary metabolites from selected medicinal plants [Syzygium cuminii (L) skeel (2 constituent), Momordica charantia Linn (6 constituent). And Cassia auriculata Linn (5 constituent) roots] in rigorous way. the pharmacological data of the crude as well as isolated constituent pioneer, the research investigation to the path of Retrosynthetic analysis (disconnection approach).

The molecule (Code McReX-1) is selected for the total synthesis. It is having highly anti diabetic, as well as immunomodulatory activity, in future will carry out anti-cancer and anti HIV activity of this synthetic product.

CONCLUSION:

The McReX-1, chemically it is named as 2-(2-amino-2-(2-(2-hydroxy-3-methylbutyl) octahydro pyrrolo [1,2-a] pyrazin-7-yl) ethyl)-6-ethylphenol, synthesis by adopting total synthetic strategy in four step. In the first step we synthesis, Octahydropyrrolo [1,2-a]pyrazine (1M) in a simple organic synthetic procedure, condensation and brominating the alkyl halide with piperzine as starting material. The product of the first step is utilized for the synthesis of step two 1-(hexa hydropyrrolo [1,2-a]pyrazin-2(1H)-yl)-3-methylbutan-2-ol (2 M). in the same strategic way the product of (2 M) to synthesis (3 M) 2-amino-2-(2-(2-hydroxy-3-methylbutyl) octahydropyrrolo [1,2-a]pyrazin-7-yl)ethan-1-ylium. Finally the product of (M 3) is used to synthesis 2-(2-amino-2-(2-(2-hydroxy-3-methylbutyl) octahydro pyrrolo [1,2-a] pyrazin-7-yl) ethyl)-6-ethylphenol successfully. In the protocol of the research we included all positive research finding. Moreover we faced various difficulty to approach the exact technique to transforming the structure of a synthetic target (TGT) molecule in a sequence of progressively simpler structures along the adopted pathway to reach the goal in the synthesis of McReX-1 from the plant Momordica charantia Linn. We succeeded to achieve the goal probably.

PHARMACOLOGICAL STUDY:

Assessment anti-hyperglycemic activity on synthesis synthetic McReX-1 in STZ induced model:

The bioactive molecule that is isolated from Momordica charentia linn is having similar physical, chemical and spectral property like glibencalimide hence we planned to developed the Synthetic procedure, by Retrosynthetic analysis, the synthetic molecule was (McReX-1) assed for anti-hyperglycemic activity in STZ induced diabetic rat model. to prove whether or not is having capacity to reduced the hyperglycemia by comparing with standard Glibencalimide

Antihyperglycemic Activity of Synthetic McReX-1 :

Animals:

Adult male Wistar albino rats weighing 170-250 g, obtained from Laboratory Animal Centre, Department of Pharmacology, NGSMIPS, Deralakatte, NITTE University Mangalore, India were used in the present study. They were housed in a clean polypropylene cage with not more than four animals per cage and maintained under standard laboratory conditions (temperature 25 ± 2 ºC with 12/12 h dark/light cycle). They were fed standard pellet diet (Hindustan Lever, Kolkata, India) and water ad libitum. The animals were acclimatized to laboratory conditions for one week prior to the experiment. All procedures described were reviewed and approved by the University Animal Ethics Committee, NITTE University Mangalore.

Acute Toxicity:

The acute oral toxicity of synthetic McReX-1 in male Swiss albino mice was studied as per OECD guideline 425 (OECD Guidelines 425 208). The median lethal dose (LD50) value was determined using the method of maximum likelihood.

Oral glucose tolerance test:

The oral glucose tolerance test was performed in overnight fasted normal rats. Rats were divided into three groups (n=6).

Group I:

Served as normal control and received distilled water (5 mL/kg b.w. p.o)

Groups II:

Received McReX-1 50 mg/kg b.w

Groups III:

Received McReX-1 100 mg/kg b.w.,

Respectively, after these treatments, all groups received glucose (2 g/kg b.w.) orally. Blood was withdrawn from the tail vein just prior to and 30, 60, 120 and 240 min after oral glucose administration (Soltani N, 2007). Blood glucose levels were measured using single touch glucometer (Accu-check, Roche Diagnostics, USA) refer table number 59 for results.

Induction of Diabetes:

Diabetes mellitus was induced in overnight fasted rats by a single intraperitoneal STZ injection (65 mg/kg b.w.) (Ravi K, 2004). After 3 days, fasting blood glucose levels were measured and the animals showing blood glucose level ≥225 mg/dL were used for the study (Ewart RBL,1975).

Treatment schedule and estimation of fasting blood glucose (FBG) level:

The rats were divided into five groups (n=6). Except for group I, which served as normal non-diabetic control, all other groups were comprised of diabetic rats.

Group I: Normal group

Group II:-served as diabetic (STZ) control.

Groups III: Received 50 mg/kg b.w., p.o.

Groups IV: - received 100 mg/kg b.w., p.o.,

Group V: received reference drug glibenclamide 0.5 mg/kg b.w., p.o. daily for 15 days. Fasting blood glucose was measured on days 0, 5, 10 and 15 by using a one-touch glucometer (Accucheck ®). At 24 h of the last dose, blood was collected from overnight fasted rats from each group by cardiac puncture for estimation of serum biochemical parameters.

Body Weight

Body weight of rats from each group was measured on days 1, 7 and 15 of McReX-1 treatment. Refer the table.

Effects of McReX-1 on fasting blood glucose levels in streptozotocin (STZ) induced diabetic rats.

Group	Treatment		Fasting blood glucose level (mg/dL)
Group	Treatment		Basal value	5^th day	10^th day	15^th day
A.	Normal Control		87.55 ± 2.9	80.91 ± 2.92	82.32 ± 1.69	80.19 ± 3.5
B.	STZ (Vehicle)		272.9 ± 3.12	285.92 ± 1.0	267.9 ±1.11	299.42 ± 5.0
C.	STZ + Glibenclamide (5 mg/kg)		260.86 ± 6.92 ( 63% )	140.25 ± 7.06*** (50%)	100.00 ± 4.5*** (41%)	80.44 ± 6.20*** (36%)
D.		STZ + McReX-1 (50 mg/kg)	210.23 ± 5.91 (62%)	130.12 ± 6.62*** (50%)	98.66± 9.09*** (39%)	78.11 ± 8.3*** (33%)
E.	STZ + McReX-1 (100 mg/kg)		200.23 ± 5.91 (62%)	120.23 ± 5.91 (62%)	82.03 ± 5.91 (62%)	78.33 ± 9.1 (62%)

Data are expressed as mean ± SEM (n=6); *P<0.001 compared with normal saline control and **P<0.001 and #P<0.01 compared with STZ control group. Gliben: glibenclamide

Effects of McReX-1 on body weight in streptozotocin (STZ) induced diabetic rats

Group	Treatment	Mean body weight (g)
Group	Treatment	Basal value	7^th day	15^th day
A.	Normal Control	180.55 ± 4.91	183.91 ± 2.92	188.32 ± 2.9
B.	STZ (Vehicle) 65mg/kg	182.91 ± 3.6	150.92 ± 5.04	136.9 ± 5.4
C.	STZ + Glibenclamide (5 mg/kg)	200.86 ± 6.92 ( 63% )	140.25 ± 7.06*** (50%)	122.18 ± 6.5*** (41%)
D.	STZ + McReX-1 (50 mg/kg)	183.11 ± 6.1 (62%)	140.12 ± 6.62*** (50%)	116.67± 9.09*** (39%)
E.	STZ + McReX-1 (100 mg/kg)	198.23 ± 6.9 (62%)	189.12 ± 6.62*** (50%)	191.67± 9.45*** (39%)

Data are expressed as mean ± SEM (n=6); *P<0.001 compared with normal saline control on corresponding day; **P<0.001 and #P<0.01 compared to STZ control group on corresponding day. Gliben: glibenclamide. Fasting blood glucose level in STZ induced rate model

Reduction of body weight in STZ induced diabetic rate modle in McReX-1

CONCLUSION:

We would be able to got successes to trace out three orally active principles called McReX-1 which are in-depth assessed for its safer and more effective compound with all the desired parameters of a drug, that could replace the synthetic medicines. This bioactive principle we have picked out the bioactive molecule McReX-1 assed it for anti-hyperglycemic activity in STZ induced diabetic rat showed the reduction glucose to normal similar to Glybincalimide which stimulated us to develop the Retrosynthetic model which have gave a meaning full conclusion to the entitled project the summery of overall conclusion of entire research project is given above.

ACKNOWLEDGEMENT:

The author sincerely thanks, the management of Anjuman-I-Islam’s New Panvel, Navi Mumbai, Director of KTC, Dr. Abdul Razak Honnutagi, and Dean Dr. Rajani Shettigai, for providing us the opportunity to write this review in a peaceful environment.

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Received on 31.01.2015 Modified on 25.02.2015

Asian J. Research Chem 8(5): May 2015; Page 299-306

DOI: 10.5958/0974-4150.2015.00050.4