Synthesis of Novel α-Amino Acids Bearing 1 , 2 , 4-triazinone and Steroidal Moieties as Enzymetic Affect ( Cellobiase Activity ) Part I

An imperative class of α-amino acids bearing 1,2,4-triazinone and N,C-disubstituted glycine (5a,b) and/or the related systems (6 and 7) have been synthesized by the condensation of 6-(2'-aminophenyl-4-phenyl-3-thion -1,2,4-triazin-5-one (1) with the appropriate steroids, Epiandrosteron (2a) and Dehydrosterone (2b), followed by the addition of hydrocyanic acid. Nucleophilic substitution of mercapto group of (5) by 4-fluoroaniline and sulfanilamide give the corresponding systems 6 and 7. Compounds 6,7, 5, and 4 showed a high enzymatic effect as cellobiase agents against some tested fungi .

Selective installation of fluorine atom into a therapeutic or diagnostic molecule can enhance a number of pharmacokinetic and physicochemical properties (Delpon, 2008), such as improved metabolic stability and enhanced membrane permeation (Shah & Westwell, 2007;Hagmann, 2008).An increased binding affinity of fluorinated drug candidates to target protein has also been reported (Filler & Saha, 2009).

Biologicalevaluation
The effects of the synthesized α-amino acid derivatives on the cellobase activity were studied using the Reese and Mandel procedure (Abdel-Aziz et al., 1996 ), (PH 5, incubated at 40 o C for 1hour).The released reducing sugar was estimated calorimetrically at 540 nm (Ibrahim et al., 1997;Abdel-Rahman, Morsy, Allimon & Abd El-Monem, 1999;Ibrahim et al., 2009), Table 1.Compounds 6a and 6b both showed a higher amount of activity over the compounds when compared against the other tested fungi, thus, the introducing of fluorine atoms and / or sulfa-drug moiety resulted in high order of activity in comparison with the corresponding α-amino acids.Additionally, the presence of fluorine substituted α-amino acids bearing 1,2,4-trizine and steroidal moieties led to increases the net-electronegativity, which improve the dielectric constant and enhances the hydrophobic properties.These properties, in all, increase their efficiency as enzymetic parameters.
In conclusion, the relationship between structural parameters, electronic parameter, and antifungalenzymatic activity ensures that the replacement of SH group of 1,2,4-triazine by substituted fluorine enhance the overall enzymetic effects.

General Procedures
The melting points were determined using a Gallenkamp apparatus and were uncorrected, IR spectrum were recorded with FT-IR Bomem MB 104 using nujol mults and NaCl cells, NMR spectrums were obtained on Brukes Avance 400MHz.chemical shift expressed in  (ppm) using DMSO-d 6 , Mass spectrum were measured on GCMS Q1000-Ex at 70eV.Microbiological analyses were performed by the microanalytical center at Ain-Shams University, Egypt.

Conclusion
This study showed that the presence of fluorine atoms and /or sulfa-drug moiety, combined with α-aminoacids, increases the cellobiase activity, while the carbonitrile derivatives decrease the tested bioactivity over the synthesized amino acid.
In addition, the incorporation of 5-ox-1,2,4-triazin-3-thione and a type of steroids to amino acid (glycine) initiatesthe potency of the novel synthesis systems, leading to the inhibition.It also accelerated its enzymatic affects.

Table 1 .
effect on cellobiase activity produced by Aspergillus nidulans Aspergillns niger fungi