Behavior of Adrenal Ectomized Rats in Chloro-Hydroxy Derivatives of Phenothiazine

INTRODUCTION

Phenothiazines are mild cholinesterase inhibitors and may therefore enhance the action of depolarizing neuromuscular blocking drugs1 and ester- linked local anesthetics2. Derivatives of Phenothiazine are highly bioactive and have widespread use and rich history. Phenothiazine derivatives are associated with potent biological activities such as anti-inflammatory3, analgesic4, insecticidal5, fungicidal6, bactericidal7 and trypanocidal8, anticonvulsant9, diutatic10, antiallergic11 and neuroleptic12. Chloro- hydroxy derivatives of phenothiazine is 8-chloro, 3-hydroxy, 10-H phenothiazine 2-carboxyalic acid on treated with chloroacetyl chloride afforded N10-chloroacetyl, 8- chloro, 3-hydroxy phenoyhiazine 2 - carboxyalic acid I. Which on reaction with hydrazine hydrate resulted in the formation of N10 – Hydrazinoacetyl, 8-chloro, 3-hydroxy phenothizine 2-carboxyalic acid II. The compound II which on condensation with various substituted aromatic carbonyls yielded substituted N10 -[α-(Benzylidene hydrazino) acetyl ] 8-chloro,3- hydroxy phenothiazine 2-carboxyalic acid III. The compound III on reaction with thioglycolic acid underwent dehydrative annulations afforded 2- substituted aryl 3-N10 [ (acetylamine) 1- 3 thiazolidin 4- ones] 8- chloro 3-hydroxy phenothiazine 2- carboxyalic acid IV.

Carrageenan is a natural polysaccharide obtained from edible red seaweeds. Carrageenan induced inflammation test13 is widely used to investigate anti inflammatory activity of any compound and constitutes a simple and routine animal model for evaluation of pain at the site of inflammation, without any injury or damage to the inflamed tissue. Carrageenan induced rat paw oedema male wister rats (g) divided into 6 animals in each groups used for study induced by subcutaneous injection of a 0.1 ml of 1% freshly prepared carrageenan in saline in the right hind paw of rats and sub- plantar- injection of histamine at dose of 0.1ml of 1%. Carrageenan solution (0.1ml/paw) was injected subcutaneously into planter surface of the right hind paw of rats. The paw volume of rats in control, standard and test groups was measured with the help of the Plethysmometer during the interval of 30, 60, 120, 180, 240 min.

Synthesis of N10 - chloroacetyl, 8- chloro –hydroxy – phenothiazine - 2- carboxyalic acid (I) → A mixture of 8-chloro, 2-hydroxy, 10-H phenothiazine 2-carboxyalic acid (3.0ml) on electrophilic substitution by chloroacetyl chloride (0.5ml) was refluxed on a steam bath for about 10 hr. The solid thus obtained was dried , purified and crystalized from ethanol (yield 60%), M.P. 140- 1440C and Calcd for C15H9NSCI2O4 : C,61.32; H,5.46; N,4.25%; Found : C,1.38; H,5.40; N,4.23%; IR 1738(>C=0), 2175 and 2236 (>N-C=0), 3035, 1575, 1550, 1475, 870 (Phenothiazine nucleus), 764(C-CI) and 690 (C-S-

Synthesis of N10-Hydrazinoacetyl, 8- chloro, 3- hydroxy phenothiazine 2- Carboxyalic acid (II) → A mixture of (0.5ml) 1 and hydrazine hydrate (0.8ml) on methanol (40ml) was refluxed on a stream bath for about 10 hr. Methanol was then removed under reduced pressure and the solid thus obtained was dried, purified and crystallized from ethanol (yield 78%) , M.P. 160-1630C ; And Caled for C15H12N3SC104 : C,62.47; H,4.47 ; N, 9.83%; Found: C,62.87; H,4.62; N,9.54% . IR: 3033, 1558,860 (phenothiazine nuclear), 3336 and 2885 (-CH2NH), 3349(NH-NH2), 2237 and 1782 (>N-C=0) and 712(C-S-C). 1H-NMR : 4.36 (S,2H-CH2), 4.44( S,2H,-NH2), 7.72 (S,1H,-NH). Synthesis of N10 - [α-(Benzylidenohydrazino) acetyl 8-chloro, 3- hydroxy phenothiazine 2-carboxyalic acid (III). → A mixture of compound 2(2.0ml) and substituted aromatic (0.5ml) with 4.5 drops of glacial acetic acid was refluxed on a water bath for about 12 hrs. Cooled, evaporated to obtain a residue and was purified. The product was recrystallised from methanol (yield 80%), M.P. 142-1460C. Anal. Calcd. For C22H15N3SO4-R1 : C,68.74; H,4.37; N,9.65% Found: C,69.58; H,4.33; N,9.67, IR: 3032, 1545, 870, 735 ( Phenothiazine and benzene nuclei) , 3340 and 2887 (-CH2NH) , 2272 and 2235 (>N-C=0) and 1546 (N=CH). 1H-NMR : 4.38 (S,2H-CH2), 4.93 (S,1H, N=CH), 7.05-8.08 (m, 13H, Ar-H). Synthesis of 2- Substituted aryl –3-N10 [(acetylamino) 1,3- Thiazolidin, 4- ones] 8- chloro, 3-hydroxy phenothiazine 2- carboxyalic acid (IV). → A mixture of compound 3 (2gm) in methanol and thioglycolic acid (1.15gm) with a pinch of anhydrous ZnCl2 was refluxed for about 10 hours on a water bath. The separated solid was filtered purified and Crystallised form methanol (yield 80%), M.P. 143-1450C, Anal Caled for C24H17N3S2O5-R1 : C,62.39; H,4018; N,8.67% Found: C,63.37; H,4.15; N,8.64%; IR : 3037, 1569, 870, 707 (Phenothiazine and benzene nuclei), 3420, 3340 and 2887 (-CH2NH), 2240 and 1785 (>N-C=0) and 1710 (C=0 cyclic) 1H-NMR : 3.15 (S,1H,-N-CH), 3.52 (S,2H,-CH2-S), 4.38(S,2H,-CH2), 7.12 – 8.10 (m, 13H Ar-H).

This section includes the result and discussion of anti-inflammatory activity of the synthesized products derived from phenothiazine. Some of the products exhibited pronounced biological activity. Structure of the synthesized compounds has been elucidated on the basis of their elemental analyses and spectral data.

We are grateful to the Head R.S.T.C. CDRI, Lucknow for providing spectral and analytical data of the compounds. We are thankful to the RRL, Bhopal for providing the platform of the Screening test. I also thankful to department of Research and development, Madhyanchal Professional University Bhopal for synthesized compounds. Cohen and Staney B. Prusiner (2001). PNAS, 98 (17), Pg. 9836-9841. 2. Sinha Sweta, Pandeya S.N., Verma Anupam, Yadav Deepika (2011). IJRAP, 2(4), Pg. 1130-1137. 3. Mishra, M. Srivastava, S. K. and Srivastava, S. D. (1997). Indian J.Chem.,36B, Pg. 826. 4. Hirata, T. Driscall, J.S. (1976). J. Pharm Science, 65, Pg. 1996. 5. Shukla, Y. K. (1994). Indian J. Chem. 3B, Pg. 799. 6. Lodhi, R. S., Srivastava, S. D. (1997). Indian J. Chem., 36B, Pg. 947. 7. Vincent,J.G. and Vincent H.W. (1955). Proc. Soc. Expt. Bio. Med., 55, Pg,112. 8. Chaurasia,S. and Srivastava, S.D. (1991). J. Indian Chem. Soc., 68, Pg. 106. 9. Trivedi, P. B. Undavia, N. K. Dave, A. M., Bhall K.N. and Desai, N. K. (1993). Indian J. Chem. Pg. 760. 10. Selvendran S., Brindha J. Vasavi C.S. (2018). Wiley online library, Vol.3, Issue 46. 11. Pluta krystian, Jelen M., Morakmlodawska Beata (2017). European Journal of Medicinal Chemistry, 138, Pg. 774-806. 12. Strazadala, L., Fiedorowicz Anna, Wvsokinska Edyta Ziolo Ewa. (2018). Molecules.,23 (10), Pg. 2443. 13. Winter, C. A., Risley, E. A., and Nuss,G. W. (1962). Proc. Sco. Expt. Bio. Med., III, Pg. 544.

Department of Chemistry, Madhyanchal Professional University, Bhopal (M.P.) dr_vikrantjain@yahoo.co.in