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TITLE
CERTIFICATE 1
CERTIFICATE-2
DECLARATION
ACKNOWLEDGEMENT
CONTENTS
ABBREVIATIONS AND NOTATIONS
1 INTRODUCTION
2 REVIEW OF LITERATURE
2.1 Introduction
2.2 Microbial Transformation of Steroids
2.3 Microbial transformation of steroidal alkaloids
2.4 Microbial Transforalption of Aikaloids
2.4.1 Tropa alkaloids
2.4.2 Pyridine alkaloids
2.4.3 lsoquinoline alkaloids
2.4.4 Ergot alkaloids
2.4.5 Rauwolfia alkaloids
2.4.6 Colchicine
2.4.7 Strychnos alkaloids
2.4.8 Vinca alkaloids
3 BIOCONVERSION STUDIES
PART 1 VINBLASTINE
3.1 Materials and methods
3.1.1 Isolation of vinblastine and vincristioe from Vim rosea
3.1.2. Isolation of vinblastine from commercially available Velbrn
3.1.3. Isolation of vincristine from commercially available Oncovin
3.1.4 Microbial conversion
3.1.4.1 Source of Microorganism
3.1.4.2 NCIM Cultures
3.1.4.3 Maintenance and preservation of NCIM cultures
3.1.4.4. Screening of NCIM cultures
3.1.4.5 Screening of microorganisms from the dung of cow and goat
3.1.4.6. Saeening of microbes from the gut and lumen of cow collected frombutchers shop
3.1.4.7 Screening of microorganisms from soil sample
3.1 5. Identification of the microorganism
3.1.5.1 Morphological characters
3.1.5.2 Biochemical characters
3.1.6 Growth studies of the isolated microorganism
3.1.6.1 Medium
3.1.6.2 Preparation of Inoculum
3.1.6.3 Incubation methods
3.1.6.4 Measurement of Growth
3.1.6.5 Effect of vinblastine sulphate on growth
3.1.6.6 Effect of pH on growth
3.1.6.7 Effect of Temperature on growth
3.1.6.8 Effect of sodium chloride on growth
3.1.6.9 Transformation of vinblastine
Analytical Methods
a) Thin Layer Chromatography (TLC)
b) High Pressure Liquid Chromatography (HPLC)
c) Estimation of vinbiastine Sulphate by H P L C ~
d) Estimation of vinblastine monohydmzide by HPLC
3.1.7. Preparation of vinblastine monohydrazide synthetically
3.1.7.1 Rate of transformation of vinblastine to vinblastinemonohydrdde
3.1.7.2 Isolation of vinblastine monohydrazide
3.1.8 Results
3.1.8.1 Introduction
3.1.8.1a Confirmation f the product as vinblastine monohydrazide byderi~rtization~
3.1 8.2 Growth studies
3.1.8.2a Fermentation of vinblastine monohydrazide
3.1.8.2b Effect of vinblastine on growth
3.1.8.2c Effect of pH on growth
3.1.8.2d Effect of temperature on growth
3.1.8.2e Effect of sodium- chloride on growth
3.1.8.3 Rates of transformation of vinblastine to vinblasthemonohydrazide
3.1.9 Discussion
Fig. 3.1.6 Rate of transformation of vioblastine to vinblastine monohydnride
PART 2 CONESSINE
3.2 Materials and methods
3.2.1 Isolation of concssine (2) from Hohrrhena antidyseentrica (Kodakapala): the whole plant
3.2.1.2 Preparation of derivative, dihydrochloride monohydrate ofconessine. 46
3.2.2 Microbial conversion
3.2.2.1 Source of microorganism
3.2.2.2 NCIM cultures
3.2.2.3 Maintenance and preservation of NClM cultures
3.2.2.4 Screening of NCIM cultures
3.2.2.5 Isolation of microbes from the dung collected from cow and goat
3.2.2.6 Isolation of microbes from the gut and lumen of cow
3.2.2.7 Isolation of microbes from soil samples
3.2.3 Identification of the isohted microorganism
3.2.3.1 Morphological characters
3.2.3.2 Biochemical characters
3.2.4 Growth studies of the isolated organism
3.2.4.1 Medium
3.2.4.2 Preparationof inoculum
3.2.4.3 Incubation procedure
3.2.4.4 Measurement of growth
3.2.4.5 Effect of wnessine on growtb
3.2.4.6 Effect of pH on growth
3.2.4.7 Effect of temperature on growth
3.2.4.8 Effect of sodiam chloride on growth
3.2.5 Effect of physicochemid factors on the conversion of conessine
3.2.5.1 Medium
3.2.5.2 Inoculation and incubation procedure
3.2.5.3 Analytical procedure
a. Extraction procedure
b. Thin Layu Chroautognphy
c. Estinution of wnessine
3.2.5.4 Effect of Sodium chloride on the conversion of wnessine
3.2.5.5 Effect of gluwse on the degradation of wnessine
3.2.5.6 Effect of pH on the conversion of wnessine
3.2.6 Isolation of 7-α-hydroxy conessine
3.2.6.1 Rtpurtion of derivative (Ketone of 7-a-hydroxy concssh) 214
3.2.7 Results
3.2.7.1 Growtb studies
3.2.7.2 Effect of conessinc on growtb
3.2.7.3 Effect of pH on growth
3.2.7.4 Effect of temperature on growth
3.2.7.5 Effect of sodium chloride on growth
3.2.8 Effect of physicochemid factors on conversion of wnessine
3.2.8.1 Effect of sodium chloride on the conversion of conessine
3.2.8.2 Effect of glucose on the conversion rate of conessine
3.2.8.3 Effect of pH on the degradation of conessine
3.2.8.4 Confirmation of the product as 7-a-hydroxy conessine
3.2.8.5 Fermentation of 7-a-hydroxy wnessine
3.2.9 Discussion
4 PHARMACOLOGICAL STUDIES
PART I VINBLASTINE AND ITS BIODEGRADATION PRODUCT
4.1 Introduction and review of literature
4. 1a Solid tumor reduction studies
Materials and Methods
Experimental animals
Maintenance of tumour cell lines
Vinblastine Preparations
Antineoplastic activity of vinblastine and vinblastine monohydrazide
Drug therapy through intrapexitone-l route
Results and Discussion
4. 1b Biochemical studies
Materials and Methods
I. Carbohydrate Metabolism
Estimation of blood glucose
II. Lipid Metabolism
1. Extraction of suum and tissues for Lipid estimation
(a) Extraction of serum
(b) Extraction of tissues for lipid estimation
2. Estimation of cholesterol
3. Estimation of higlycerides
4. Estimation of phospholipids
III. Protein Metabolism
1. Estimation of protein
2. Estimation of blood urea
IV. Glycoprotein metabolism
1. Extraction of glycoproteins from the tissues
(a) Preparation of dry defatted tissue
(b) Papain digestion
2. Estimation of total hexose
3. Estimation of fuwse
4. Estimation of sialic acid
V. Assay of serum enzymes
1. Estimation of serum lactate dehydrogenase
2. Estimation of serum transaminases
3. Serum phosphatases
VI. Statistical analysis
Results and Discussion
1. Gain in weight
2. Carbohydrate metabolism
Blood glucose estimation
DISCUSSION
3. Lipid metabolism
1. Cholesterol
2. Triglycerides
3. Phospholipids
4. Protein metabolism
1. Total protein
2. Blood area
5. Glywprotein metabolism
1. Protein bound hexose
2. Protein bound fucose
3. Protein bound sialic acid
6. Serum enzymes as tumor markers
1. Serum lactate dehydrogenase
2. Serum transaminases
3. Serum phosphatases
PART 2 CONESSINE AND ITS BIODEGRADATION PRODUCT
Introduction and review of literature
Materials and Methods
Experimental animals
Conessine preparations
Induction of diarrhoea
Results and Discussion
5 SUMMARY OF IMPORTANT FINDINGS
REFERENCES
Fig. Caption