Study the effect of oxazepine derivatives of alkaline phosphatase in normal persons sera

: The inhibitory effect of new organic compound which is derivative of oxazepine on the activity of alkaline phosphatase in normal persons serum have been studied This compound was :

The percentage of inhibition caused by oxazepine derivative was to be arranged between 22.5 -69 % The result from line weaverburk plot indicated that the inhibition was competitive which maximum velocity vmaxwhile Micheal menten constant Km is decreased in presence of inhibitor 9 m mol/L while the value of Km without inhibitor is 10 m mol/L .synthesis new 1,3 oxazepine 4-7 dion derivatives which are expected to have biological activity which are most active anticonvulsant.

Introduction
The alkaline phosphatase are agroup of enzymes which hydrolyze organic phosphatase at high PH .They are present in most tissue but are in particularly high concentration in osteoblasts of bone , the cells of the hepatobilliary tract , the intestinal wall , the renal tubules and placenta 1 , the molecular weight varies with the tissue source of enzyme and ranges from 70.000 to 120.000 adlton 2 .
ALP is a family of dimeric metalloenzymes and requires Mg +2 and Zn +2 for stability and maximum activity .ALP is group of nonspecfic phosphates that catalyze the reaction as shown below :

Oxazepine compound :
Oxazepine is non-homologous seven membered ring that contain two hetro atoms oxygen and nitrogen 1,3 oxazepine Oxazepinedione prepared from shiff's bases with selected anhydride

Materials and chemicals
Phosphatase alkalinkit biomerieux company / ethanol 99% BDH company / England Sample : the samples were collected from the bank of blood of the normal persons Principle : The activity of ALP was measured by 7,8 employ a colorimetric method to the following reaction : The liberated phenol is measured in the presence of 4-amino and pyrine and potassium ferriccyanid

Determination of ALP activity before and after addition of inhibitor
The activity of ALP enzyme in sera of normal persons without inhibitor was determined according to (kind and Belfield etal )   The oxazepine compound moiety showed liquid crystal lyotropic type result from change in concentration of solution.When the concentration increases transition from non-ordered isotropic solution to an ordered anisotropic solution can take place 11.These factors were inhibition the activity of enzyme Table 3 and fig  Explain line weaver Burk plot of ALP in normal persons sera at constant concentration of organic compound (10¯4) mMol / L with changed of substrate concentration was competitive with Vmax 6.66 µl and Km value 10 m Mol/L for uninhibited reaction and 9 m Mol/L for inhibited reaction .This organic compound is new oxazepine derivative so that no available reference.

7 , 8 5 -
The activity of ALP enzyme was calculated from following equation : Sample ALP = OD sample -OD sample blank ‫ــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــ‬ OD standard OD = absorbance = A n= 142 U/L determination of ALP enzyme activity with inhibitor (I) while the concentration of substrate (S) and enzyme were fixed 1-Aliquate 50 µl of sample was pipetted in each four test tube 2-Aliquate from inhibitor concentration (10¯3 , 10¯5 , 10¯6, 10¯7 and 10¯8 ) were placed in separate tube .3-the Steps were repeated according to (kind and Belfield etal ) 7,8. 4-The effect of inhibitor was calculated according to the following equation Inhibitor = 100 -activity with inhibitor ‫ــــــــــــــــــــــــــــــــــــــــــــــــــ‬ Activity without inhibitor 5The highest percentage of inhibition was used to obtain the type of inhibition .the type of inhibitor was determined using different concentration of substrate (6×10¯3 ,7×10¯3 ,8×10¯3 ,9×10¯3, 10×10¯3) m mol / L concentration of ( I ) and (E) were fixed .lineweaver -Burk plot was used to find the type of inhibition .

1
Showed michel menten plot for ALP in normal persons the value of vmax 10 U/L while the Km value was 58 × 10¯4 m mol/LTable 4 A,B and fig 2 :