Synthesis, Characterization and Biological Evaluation of Novel Triazolyl - Acridine Derivatives as Cytotoxic Agents

 

Mandeep Singh1*, Dr. D. N Prasad2, Dr. Supriya Agnihotri3

1Research Scholar, IKG Punjab Technical University, Kapurthala, Jalandhar, Punjab, India.

2Shivalik College of Pharmacy, Nangal, Punjab, India.

3Chandigarh College of Pharmacy, Landran, Punjab, India.

*Corresponding Author E-mail: mandeepchadha7@gmail.com

 

ABSTRACT:

Acridine and Triazols both are biologically active heterocyclic rings with cytotoxic potential. Triazolyl- acridine adduct attract the attention in the field of medicinal chemistry. Here we synthesized a series of triazolyl- acridine compounds by the appropriate procedures. Structure of all synthesized compounds was confirmed by the various spectroscopic methods e.g. FT-IR, proton NMR and Mass spectrograms. All synthesized triazolyl-acirdines compounds were assayed in-vitro for cytotoxic activity against MCF-7 (human breast adenocarcinoma cell line) and HT-29 (human colon adenocarcinoma cell line) cells by MTT- assay. All target compounds shows increasing activity in dose dependent manner. However MPSP-9 was sensitive against MCF-7 but compound MPSP-1 was most sensitive with minimum inhibitory concentration (IC50 value) against MCF-7 and HT-29 both cell lines.

KEYWORDS: Triazol, acridine, cytotoxic, cell line, heterocyclic.

 

 

INTRODUCTION:

Acridine is a heterocyclic organic compound containing a nitrogen hetero atom, with the formula C13H9N. Chemically, acridine is dibenzopyridine, 2,3,5,6- dibenzopyridine and 10-azaanthracene. Acridine has an irritating odor. It is also found in plants as alkaloid. The antimicrobial property of acridine discovered in 1917, by Ehrlich and Benda1. It is also characterized by irritation on skin and by the blue fluorescence showed by solutions of its salts2.

 

Fig. 1: Acridine ring structure and its numeration

Structurally, Acridines are characterized as the linear fusion of three planer aromatic 6-membered rings with a hetero nitrogen atom included in the center ring3. Acridine able to bind DNA has made it a target for anticancer and antimicrobial uses4,5. Acridine can also be named as 10-azaanthracene and dibenzo[b,e]pyridine. Structure of acridine and its numeration are shown below in Figure No. 1. Triazoles are heterocyclic compounds featuring five- membered ring of two carbon atoms and three nitrogen atoms as part of the aromatic five-membered ring6. Triazole refers to either one of a pair of isomeric chemical compounds with molecular formula C2H3N3.These may be two types7.

·           1,2,3-triazoles or v-triazoles

·           1,2,4-triazoles or s-triazoles

 

1,2,3–triazole and its derivatives enhanced considerable attention for the past few decades due to their chemotherapeutical value. 1,2,3-Triazole moieties are attractive connecting units, since they are stable to metabolic degradation and capable of hydrogen bonding which can be favorable in binding of biomolecular targets Many 1,2,3–triazoles are found to be more potent anti-microbial8, analgesic9, antiallergic, anti- convulsant10, antineoplastic11, anti-malarial, and anticancer activities12.

 

MATERIALS AND METHODS:

Synthetic grade chemicals were used for the experimental work. Thin layer chromatography (TLC) on precoated TLC plates were used to monitor the synthesis and finally prepared compounds. Chloroform: methanol ratio in 9:1 ratio was used as mobile phase for TLC and iodine chamber to visualized spots. Open glass capillary method was used for Melting points determination. Infrared (IR) spectra were recorded on Bruker’s FT-IR spectrophotometer as KBr pellet and values are expressed in cm-1. 1H nuclear magnetic resonance spectroscopy (NMR) spectral analysis of the synthesized compounds were recorded on a Bruker Avance II 400 MHz in deuterated chloroform (CDCl3) using tetramethylsilane as internal standard. The chemical shift values were recorded on δ scale. Syntheses of the triazolyl-acridine derivatives (1-12) were performed according to fig. No. 1, through following steps.

 

Step 1: Synthesis of 9-chloro acridine:

9-Chloro- acridine was synthesized by modified Ullman-

Goldberg reaction13. And

 

(a)       Synthesis of N-Phenylanthranilic Acid:

Equal mol (0.038mol) of o-chlorobenzoic acid and aniline with 0.12gm of copper powder in 40ml isoamyl alcohol then 6gm of dry potassium carbonate was added slowly, final mixture was refluxed for 6 to 8 hours. After reflux, isoamyl alcohol was removed by simple distillation then mixture poured into 500ml of hot water and filtered. The filtrate was acidified with concentrated hydrochloric acid. Yellow Precipitate formed, filtered, washed with hot water and collected.

 

(b)       Synthesis of 9-chloroacridine by cyclizetion of N- Phenylanthranilic Acid:

5gm (0.023mol) of N-Phenylanthranilic acid is mixed with 16ml (27gm, 0.176moles) of phosphorus oxychloride taken in the in a 500ml round bottom flask fitted with a water-cooled condenser and heated about 15 minutes at 85-900C, then temperature is raised to 135- 1400C, where it is maintained for 2 hours. Excess phosphorus oxychloride is removed either by simple distillation. After cooling, the residue was poured into a well-stirred mixture of 20ml of concentrated ammonia solution, 50gm of ice and 20ml of chloroform in a separating funnel and allowed to stand for 30-40 minute. Separated the chloroform layer and evaporated, greenish gray powder is obtained is 9-chloro acridine.

 

Step 2(a): Addition of alkanediols on 9- chloroacridine14:

Dissolved 0.2g (1.0mmol) of 9-chloroacridine in 2.0mL of ethylene glycol, under inert atmosphere a 1.0 M solution of potassium t-butoxide in t-butyl alcohol (1.5 mL, 1.5mmol, 1.5 equiv) was added, stirred at 80°C for 18 h, then quenched with saturated NaHCO3. The mixture was extracted with CH2Cl2, dried over anhydrous MgSO4, concentrated in vacuum, and purified by recrystallization from CHCl3 to yield 9-acridinyl alkanediols as product.

 

Step 2(a): Conversion of 9-chloroacridine to aminoalcohols:

9-chloroacridine (1.0g, 5.1mmol) and alkanolamine (5equiv) was heated at 110°C for 6 h, under inert atmosphere and then cool. Then 1N NaOH was added then extracted with CHCl3. The organic layers were washed with brine, dried with anhydrous Na2SO4, and evaporated under reduced pressure to yield 9-acridinyl alkanolamines as product.

 

Step 3: Conversion of alcohols into azides15:

Firstly, 3 mMol of 9-acridinyl alkanediols or 9-acridinyl alkanolamines, 3.6 mmol of triphenylphosphine and 3.6 mmol of iodine was triturate in pestle mortar for 10 min, when exothermic reaction took place, paste like consistency appeared. Separately, 12mmol of sodium azide dissolved in DMSO. Sodium azide dissolved DMSO solution mixed with acridine paste and stirred for 30 min on magnetic stirrer. Upon completion of the reaction, ice-cooled solution of 50mL of sodium thiosulphate was added and extracted with 30mL of diethyl ether 3 times. The combined organic layer was washed with brine (50mL). On evaporation the crude product obtained, which was purified by column chromatography using 5% ethyl acetate in hexane.

 

Step 4: Conversion of azides into triazoles16:

3mmol of Alkyne and, 3 mmol of azide were suspended in 12mL of a 1:1 water/tert-butanolmixture. Freshly prepared 1M Sodium ascorbate solution was added, followed by 0.03mmol of copper (II) sulfatepentahydrate in 100mL of water. The heterogeneous mixture was stirred vigorously overnight. When TLC analysis indicated complete consumption of the reactants, the reaction mixture was diluted with 50mL of water and cooled in ice, and the white precipitate was collected as filtrate. After being washed with cold water (225mL), the precipitate was dried under vacuum to obtained pure triazole compounds 1-12.

 

The   aim   is   to   develop   potent   triazolyl-   acridine

 

SCHEME:

 

 

Fig 2: Scheme for synthesis of target compounds

 

RESULTS:

MPSP-1: 2-methoxy-9-(2-(4-phenyl-1H-1,2,3-triazol- 1-yl) ethoxy) acridine:

Colour: Whitish yellow crystal, Yield: 81%, M.P.: 354oC, Rf: 0.55, Elemental analysis calcd. (Found)% for C24H20N4O2      C:    70.71(70.65); H:    5.08(5.13);    N:

14.13(14.02); O: 8.07(8.12). FT-IR (KBr):cm-1 3072-C- H str. (triazole), 1462-C-H str.(methylene), 1409-C-H str.(methyl), 1682-C=N str., 1576-N=N str., 1544- C=Cstr.(Ar), 1332-C-N str., 1227-C-O str.(ether).1H NMR (CDCl3, 400 MHz): δ 8.00(d, 1H, J =10.0, CH), 7.88(d, 1H, J =6.64, CH), 7.82(d, 2H, J =7.6, CH-Ar), 7.79(t, 2H, J =3.12, 4.92, CH), 7.67(s, 1H, Ar-CH), 7.62(t, 1H, J =4.2, 4.8, CH), 7.59(s, 1H, CH), 7.53(t, 2H, J =3.8, 2.04, CH-Ar), 7.45(t, 1H, J =3.04, 5.24, CH- Ar),7.35(d, 1H, J =4.88, CH), 4.51(t, 2H, J =1.92, 7.12, CH), 4.16(t, 2H, J =5.24, 5.08, CH), 3.84(s, 3H, O-CH3), ppm.MS: 397 (M+1)

 

MPSP-2: 9-(2-(4-(2-chlorophenyl)-1H-1,2,3-triazol-1- yl)ethoxy)-2-methoxyacridine:

Colour: White crystal, Yield: 85%, M.P.: 332oC, Rf: 0.52, Elemental analysis calcd. (Found)% for C24H19ClN4O2     C:   66.9(66.24);   H:   4.44(4.34);   N:

13.0(13.0); Cl:   8.23(8.12);   O:   7.43(7.31).   FT-IR (KBr):cm-1 3039-C-H str. (triazole), 1457-C-H str.(methylene), 1421-C-H str.(methyl), 1652-C=N str., 1596- N=N str.,1521- C=C str.(Ar), 1302-C-N str., 1259- C-O str.(ether), 827-C-Clstr. 1H NMR (CDCl3, 400 MHz): δ 7.99(d, 1H, J =7.72, CH), 7.88(d, 1H, J =6.8 CH), 7.78(t, 2H, J =0.88, 3.92, CH), 7.74(d, 1H, J =7.24,CH-Ar), 7.67(s, 1H, Ar-CH), 7.62(t, 1H, J =6.52, 3.36, CH), 7.59(s, 1H, CH), 7.56(d, 1H, J =7.36, CH-Ar),7.40(t, 1H, J =4.76, 4.84, CH-Ar),7.35(t, 2H, J =2.48,5.64, CH), 4.49(t, 2H, J =5.88, 3.08, CH), 4.16(t, 2H, J=4.76, 2.0, CH), 3.83(s, 3H, O-CH3), ppm..MS: 431 (M+1)

 

MPSP-3: 2-methoxy-9-(2-(4-(o-tolyl)-1H-1,2,3-triazol- 1-yl)ethoxy)acridine:

Colour: White amorphous powder, Yield: 83%, M.P.: 276oC, Rf: 0.71, Elemental analysis calcd. (Found)% for C25H22N4O2     -C:    73.15(73.03);    H:    5.40(5.14);    N:13.65(13.49); O: 7.80(7.41). FT-IR (KBr):cm-1 3072-C-H str. (triazole), 1462-C-H str.(methylene), 1400-C-H str.(methyl), 1682-C=N str., 1576-N=N str., 1544-C=C str.(Ar), 1332-C-N str., 1266-C-O str.(ether). 1H NMR (CDCl3, 400 MHz): δ 8.00(d, 1H, J =7.12, CH), 7.90(d,1H, J =7.92, CH), 7.79-7.77(m, 2H, CH), 7.67(s, 1H, Ar-CH), 7.62(t, 1H, J =2.52, 4.72, CH), 7.59(s, 1H, CH), 7.57(d, 1H, J =8.84, CH-Ar), 7.35(d, 1H, J =8.52, CH- Ar),7.32-7.27(m, 3H, CH),  4.49(t, 2H, J =3.28,  3.72,CH), 4.16(t, 2H, J =3.52, 3.24, CH), 3.83(s, 3H, O-CH3), 2.59(s, 3H, CH3)ppm.MS: 411 (M+1)

 

MPSP-4: 2-methoxy-9-(3-(4-phenyl-1H-1,2,3-triazol- 1-yl) propoxy) acridine:

Colour: yellowish powder, Yield: 89%, M.P.: 329oC, Rf: 0.62, Elemental analysis calcd. (Found)% for C25H22N4O2-   C:   73.15(73.19);   H:   5.40(5.21);   N:

13.65(13.69); O: 7.80(7.74). FT-IR (KBr):cm-1 3087-C-

H str. (triazole), 1460-C-H str.(methylene), 1400-C-H str.(methyl),1679- C=N str.,1575- N=N str., 1541-C=C str.(Ar), 1331-C-N str., 1264-C-Ostr.(ether). 1H NMR (CDCl3, 400 MHz): δ 7.99(d, 1H, J =9.24, CH), 7.88(d,1H, J =7.36, CH), 7.80-7.78(m, 4H, CH), 7.68(s, 1H, Ar-CH), 7.63(t, 1H, J =5.92, 4.56, CH), 7.59(s, 1H, CH),7.53(t, 2H, J =7.52, 3.24, CH-Ar), 7.43(t, 1H, J =6.0,3.64, CH-Ar), 7.37(d, 1H, J =7.72, CH-Ar),4.47(t, 2H, J=4.84, 5.44, CH), 4.08(t, 2H, J =4.12, 4.2, CH), 3.86(s, 3H, O-CH3), 2.19(p, 2H, CH2)ppm.MS: 411 (M+1)

 

MPSP-5: 9-(3-(4-(2-chlorophenyl)-1H-1,2,3-triazol-1- yl)propoxy)-2-methoxyacridine:

Colour: white amorphous powder, Yield: 83%, M.P.: 256oC, Rf: 0.67, Elemental analysis calcd. (Found)% for C25H21ClN4O2-   C:   67.49(67.41);   H:   4.76(4.72);   Cl:7.97(7.81);   N:   12.59(12.52);   O:   7.19(7.10).   FT-IR(KBr):cm-1 3047-C-H str. (triazole), 1449-C-H str.(methylene), 1416-C-H str.(methyl), 1642-C=N str., 1513-C=C str.(Ar), 1338-C-N str., 1232-C-O str.(ether), 794-C-Clstr. 1H NMR (CDCl3, 400 MHz): δ 7.99(d, 1H, J =10.48, CH), 7.88(d, 1H, J =7.44, CH), 7.79-7.76(m,3H, CH), 7.67(s, 1H, Ar-CH), 7.62(t, 1H, J =1.6, 4.68,CH), 7.59(s, 1H, CH), 7.57(d, 1H, J =10.96, CH-Ar),7.40-7.34(m, 3H, CH-Ar), 4.47(t, 2H, J =5.08, 4.96,CH), 4.06(t, 2H, J =5.24, 5.08, CH), 3.86(s, 3H, O-CH3), 2.19(p, 2H, CH2)ppm.MS: 445 (M+1)

 

MPSP-6: 2-methoxy-9-(3-(4-(o-tolyl)-1H-1,2,3-triazol- 1-yl)propoxy)acridine:

Colour: light yellow amorphous powder, Yield: 75%, M.P.: 287oC, Rf: 0.53, Elemental analysis calcd. (Found)% for C26H24N4O2 C: 73.56(73.51); H: 5.70(5.61);   N:   13.20(13.02);   O:   7.54(7.47).   FT-IR (KBr):cm-1 3005-C-H str. (triazole), 1461-C-H str.(methylene), 1416-C-H str.(methyl), 1616-C=N str., 1518-C=Cstr.(Ar), 1345-C-N str., 1265-C-O str.(ether). 1H NMR (CDCl3, 400 MHz): δ 8.00(d, 1H, J =7.12, CH), 7.90(d, 1H, J =7.92, CH), 7.79-7.77(m, 2H, CH), 7.67(s, 1H, Ar-CH), 7.62(t, 1H, J =4.8, 2.88, CH), 7.59(s, 1H, CH), 7.57(d, 1H, J =11.04, CH-Ar), 7.35- 7.27(m, 4H, CH-Ar), 4.49(t, 2H,  J =1.92, 7.24, CH), 4.06(t, 2H, J =3.2, 2.56, CH), 3.83(s, 3H, O-CH3), 2.59(s, 3H, CH3), 2.21(p, 2H, CH2)ppm.MS: 425 (M+1)

 

MPSP-7: 2-methoxy-N-(2-(4-phenyl-1H-1,2,3-triazol- 1-yl)ethyl)acridin-9-amine:

Colour: White crystal, Yield: 84%, M.P.: 354oC, Rf: 0.61, Elemental analysis calcd. (Found)% for C24H21N5OC: 72.89(72.76); H: 5.35(5.27); N: 17.71(17.63); O: 4.05(4.09). FT-IR (KBr): cm-1 3396-N- H str. (secondary amine), 3073-C-H str. (triazole), 1461- C-H str.(methylene), 1400-C-H str.(methyl), 1677-C=N str., 1576-N=N str., 1543-C=C str.(Ar), 1332-C-N str., 1265-C-O str.(ether).  1H NMR (CDCl3, 400 MHz): δ8.20(d, 1H, J =9.64, CH), 7.99(d, 1H, J =7.04, CH),7.82(d, 2H, J =6.76, CH), 7.79(t, 1H,J =3.32, 4.24,CH),7.74(d, 1H, J =7.24, CH), 7.63(t, 1H, J =0.72, 6.96, CH),7.58(s, 1H, CH), 7.54(t, 2H, J =2.52, 2.52, CH-Ar),7.45-7.42(m, 2H, CH), 7.29(d, 1H, J =7.2, CH), 5.61(t,2H, J =6.0, 1.68, CH), 4.03(s, 1H, NH), 3.83(s, 3H, O-CH3), 3.55(t, 2H, J = 2.2, 5.04, CH2)ppm.MS: 396 (M+1)

 

MPSP-8: N-(2-(4-(2-chlorophenyl)-1H-1,2,3-triazol-1- yl)ethyl)-2-methoxyacridin-9-amine:

Colour: yellowish Crystal, Yield: 82%, M.P.: 321oC, Rf: 0.62, Elemental analysis calcd. (Found)% for C24H20ClN5O-C:    67.05(66.98);    H:    4.69(4.61);    Cl:

8.25(8.14);   N:   16.29(16.12);   O:   3.72(3.67).   FT-IR(KBr):cm-1 3343-N-H str. (secondary amine), 3043-C-H str. (triazole), 1459-C-H str.(methylene), 1423-C-H str.(methyl), 1653-C=N str., 1513-C=C str.(Ar), 1330-C- N str., 1228-C-O str.(ether), 825-C-Clstr. 1H NMR (CDCl3, 400 MHz): δ 8.18(d, 1H, J =9.24, CH), 7.98(d,1H, J =6.8, CH), 7.79(t, 1H,J =6.04, 4.92,CH), 7.75(d,2H, J =5.36, CH), 7.62(t, 1H, J =6.4, 2.8, CH), 7.59(s,1H, CH), 7.52(d, 1H, J =2.52, CH-Ar), 7.44 (s, 1H, CH),7.40 (t, 1H,J =3.32, 2.6,CH),7.35(t, 1H, J = 4.04, 5.24,CH), 7.29(d, 1H, J =8.76, CH), 5.60(t, 2H, J =3.24, 3.2,CH), 4.00(s, 1H, NH), 3.83(s, 3H, O-CH3), 3.55(t, 2H, J=4.72, 4.76, CH2)ppm.MS: 430 (M+1)

 

MPSP-9:                  2-methoxy-N-(2-(4-(o-tolyl)-1H-1,2,3-

triazol-1-yl)ethyl)acridin-9-amine:

Colour: white crystal, Yield: 85%, M.P.: 342oC, Rf: 0.70, Elemental analysis calcd. (Found)% for C25H23N5OC:     73.33(73.23);     H:     5.66(5.41);     N:

17.10(17.01); O: 3.91(3.81). FT-IR (KBr): cm-1 3341-N-H str. (secondary amine), 3057-C-H str. (triazole), 1462- C-H str.(methylene), 1405-C-H str.(methyl), 1681-C=N str., 1522-C=C str.(Ar), 1333-C-N str., 1230-C-O str.(ether). 1H NMR (CDCl3, 400 MHz): δ 8.20(d, 1H, J =7.08, CH), 7.97(d, 1H, J =7.4, CH), 7.79(t, 1H, J =3.12,4.8,CH), 7.75(d, 1H, J =9.4, CH), 7.62(t, 1H, J =2.4,2,68, CH), 7.59(s, 1H, CH), 7.58(d, 1H, J =8.92, CH-Ar), 7.44 (s, 1H, CH), 7.33-7.27(m, 4H, CH), 5.59(t, 2H,J =3.8, 1.8, CH), 4.01(s, 1H, NH), 3.83(s, 3H, O-CH3),3.56(t, 2H, J =4.68, 4.64, CH2), 2.59(s, 3H, CH3)ppm.MS: 410 (M+1)

 

MPSP-10:                  2-methoxy-N-(3-(4-phenyl-1H-1,2,3- triazol-1-yl)propyl)acridin-9-amine:

Colour: White crystal, Yield: 81%, M.P.: 287oC, Rf: 0.71, Elemental analysis calcd. (Found)% for C25H23N5OC: 73.33(73.15); H: 5.66(5.69); N:
17.10(17.02); O: 3.91(3.79). FT-IR (KBr):cm-1 3344-N-H str. (secondary amine), 3067-C-H str. (triazole), 1458- C-H str.(methylene), 1416-C-H str.(methyl), 1683-C=N str., 1511-C=C str.(Ar), 1335-C-N str., 1262-C-O str.(ether). 1H NMR (CDCl3, 400 MHz): δ 8.22(d, 1H, J=7.08, CH), 7.98(d, 1H, J =10.44, CH), 7.82(d, 2H, J=7.24, CH), 7.79(t, 1H,J =5.24, 2.96, CH), 7.74(d, 1H, J=10.56, CH), 7.62(t, 1H, J =2.68, 7.04, CH), 7.59(s, 1H,CH), 7.52(t, 2H, J =6.48, 3.2, CH), 7.44 (s, 1H, CH),7.42(t, 1H,J =5.24, 5.08, CH), 7.30(d, 1H, J =7.36, CH),4.46(t, 2H, J =4.72, 5.2, CH), 4.01(s, 1H, NH), 3.83(s,3H, O-CH3), 3.37(t, 2H, J =3.32, 4.36, CH2), 2.57 (p,2H, CH2)ppm.MS: 410 (M+1)

 

MPSP-11: N-(3-(4-(2-chlorophenyl)-1H-1,2,3-triazol- 1-yl)propyl)-2-methoxyacridin-9amine:

Colour: Yellow crystal, Yield: 82%, M.P.: 325oC, Rf: 0.52, Elemental analysis calcd. (Found)% for C24H22ClN5O-C:    67.64(67.48);    H:    5.0(4.95);    Cl:

7.99(7.84);   N:   15.78(15.53);   O:   3.60(3.53).   FT-IR (KBr):cm-1 3391-N-H str. (secondary amine), 3066-C-H str. (triazole), 1464-C-H str.(methylene), 1413-C-H str.(methyl), 1655-C=N str., 1537-N=N str., 1505-C=C str.(Ar), 1317-C-N str., 1266-C-O str.(ether),764-C- Clstr. 1H NMR (CDCl3, 400 MHz): δ 8.20(d, 1H, J=8.24, CH), 7.98(d, 1H, J =5.84, CH), 7.79(t, 1H,J=1.16, 7.00, CH), 7.73(d, 2H, J =8.96, CH), 7.63(t, 1H, J=3.6, 5.32, CH), 7.58(s, 1H, CH), 7.55(d, 1H, J =6.88,CH), 7.44 (s, 1H, CH), 7.40(t, 1H,J =5.88, 4.2, CH),7.36(t, 1H, J =3.6, 4.04, CH),7.28(d, 1H, J =9.04, CH),4.47(t, 2H, J =0.72, 7.24, CH), 4.01(s, 1H, NH), 3.83(s,3H, O-CH3), 3.36(t, 2H, J =4.96, 4.48, CH2), 2.59 (p,2H, CH2)ppm.MS: 444 (M+1)

 

MPSP-12:               2-methoxy-N-(3-(4-(o-tolyl)-1H-1,2,3-

triazol-1-yl)propyl)acridin-9-amine:

Colour: white amorphous powder, Yield: 86%, M.P.: 289oC, Rf: 0.63, Elemental analysis calcd. (Found)% for C26H25N5O-C:     73.74(73.61);     H:     5.95(5.83);     N:16.54(16.47); O: 3.78(3.78). FT-IR (KBr): cm-1 3343-N-H str. (secondary amine), 3067-C-H str. (triazole), 1460- C-H str. (methylene), 1401-C-H str.(methyl),1687-C=N str., 1541-N=N str., 1511-C=C str.(Ar), 1317-C-N str., 1263-C-O str.(ether). 1H NMR (CDCl3, 400 MHz): δ8.22(d, 1H, J =8.08, CH), 7.98(d, 1H, J =8.72, CH),7.79(t, 1H,J =4.04, 3.84, CH), 7.74(d, 1H, J =5.4, CH),7.63(t, 1H, J =6.12, 0.96, CH), 7.59(s, 1H, CH), 7.58(d,1H, J =9.84, CH), 7.44 (s, 1H, CH), 7.34-7.28(m, 4H,CH),4.46(t, 2H, J =1.16, 4.68, CH), 4.01(s, 1H, NH),3.83(s, 3H, O-CH3), 3.37(t, 2H, J =4.96, 1.16, CH2), 2.59 (p, 2H, CH2)ppm.MS: 424 (M+1)

 

Biological Evaluation:

All the newly synthesized triazolyl- acirdines were assayed in-vitro for cytotoxic activity against MCF-7 (human breast adenocarcinoma cell line) and HT-29

(human colon adenocarcinoma cell line) cells were maintained at 37 °C in a humidified atmosphere (90%) containing 5% CO217.

 

MTT Assay:

All the newly synthesized triazolyl-acirdines compounds were dissolved in DMSO and serially diluted with complete medium to get the concentrations a range of test concentration. DMSO concentration was kept <0.1% in all the samples. HT-29 colon carcinoma and MCF7 cells Breast adenocarcinoma maintained in appropriate conditions were seeded in 96 well plates and treated with different concentrations of the test samples and incubated at 37°C, 5%CO2 for 96 hours. MTT [(3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide)] reagent was added to the wells and incubated for 4 hours; the dark blue formazan product formed by the cells was dissolved in DMSO under a safety cabinet and read at 550nm. Percentage inhibitions were calculated and plotted with the concentrations used to calculate the IC50 values. The results of MTT Assay summarized in table no. 1 and table no. 2.

 

Table 1: Cytotoxic Activity of Synthesized Compounds (1-12) as Percentage Inhibition Against MCF7 Cell Line

S.

No.

Compound code

Concentrations

IC50

value

(µM)

0.001 (µM)

0.01 (µM)

0.1 (µM)

1 (µM)

10 (µM)

1

MPSP-1

1.8

12.48

22.19

35.65

68.95

1

2

MPSP-2

2.8

11.8

12.69

25.84

32.65

>10

3

MPSP-3

1.22

1.25

2.32

5.62

33.84

>10

4

MPSP-4

2.14

3.65

10.26

21.55

28.65

>10

5

MPSP-5

1.14

1.22

3.28

10.65

42.15

>10

6

MPSP-6

1.25

3.36

12.56

18.56

24.15

>10

7

MPSP-7

2.58

3.32

11.36

12.54

35.64

>10

8

MPSP-8

1.74

3.26

10.28

25.64

32.62

>10

9

MPSP-9

1.95

3.33

10.85

51.30

65.32

1

10

MPSP-10

1.54

3.54

12.22

18.56

32.85

>10

11

MPSP-11

1.35

4.32

12.4

21.43

30.32

>10

12

MPSP-12

1.65

2.54

10.23

19.22

25.54

>10

represented in % inhibition, IC50 value also calculated by MTT- Assay.

 

Table 2: Cytotoxic Activity of Synthesized Compounds (1-12) as Percentage Inhibition Against HT-29cell Line.

S.

No.

Compound code

Concentrations

IC50

value

(µM)

0.001 (µM)

0.01 (µM)

0.1 (µM)

1 (µM)

10 (µM)

1

MPSP-1

2.25

12.25

31.59

42.65

66.35

1

2

MPSP-2

3.25

12.54

13.65

41.25

45.26

>10

3

MPSP-3

1.55

1.23

10.23

12.54

32.22

>10

4

MPSP-4

3.35

10.31

11.56

21.36

32.26

>10

5

MPSP-5

2.26

6.54

10.26

12.65

44.26

>10

6

MPSP-6

1.25

3.36

12.56

18.56

24.15

>10

7

MPSP-7

3.67

12.26

18.65

38.56

41.26

>10

8

MPSP-8

5.62

7.46

15.63

25.69

37.26

>10

9

MPSP-9

3.41

10.23

15.33

21.62

37.22

>10

10

MPSP-10

1.25

6.26

18.5

22.16

43.62

>10

11

MPSP-11

2.34

5.32

10.43

21.43

32.43

>10

12

MPSP-12

1.65

2.54

9.24

19.44

27.54

>10

represented in % inhibition, IC50 value also calculated by MTT- Assay.

 

DISCUSSION:

Structures of novel synthesized triazolyl-acirdines compounds 1-12 were confirmed by FT-IR, 1H NMR & Mass spectroscopy data as well as their distinct Rf values in TLC analysis. FT-IR spectroscopic data clearly confirms the formation of target compounds, by showing stretching of triazole’s C-H bond at 3000cm-1, methylene’s C- H bond about 1465 cm-1, aryl alkyl ether’s C-O between 1200-1275cm-1 (1-6 compounds), secondary amine’s N-H bond between 3310-3350cm-1 (7-12 compounds) and disappearance of azide’s peaks found between 2120-2160 cm-1. Similarly proton NMR signal characterized the target compounds by δ-value of, CH at triazole ring at 7.59ppm, CH2 ranges 2.1- 5.5ppm and NH at 4.01ppm (compound 7-12).

 

All novel synthesized triazolyl-acirdines compounds (1- 12) were evaluated in-vitro for cytotoxicity against MCF-7 (human breast adenocarcinoma cell line) and HT-29 (human colon adenocarcinoma cell line). IC50 values were calculated by MTT assay, using five different concentrations of all triazolyl-acirdines compounds. MCF7 cell line was found sensitive against MPSP-1and MPSP-9, with IC50 values at 1µM. HT-29 cell line was found sensitive against MPSP-1with IC50 values at 1µM. The table no.1 and table no.2 clearly shows the inhibition of growth of MCF7 and HT-29 cells which decreases with increasing of dose administered. At the final dose, there was only minimal amount of cells survived.

 

CONCLUSION:

This study described that novel synthesized triazolyl- acirdines compounds (1-12) were inhibited the growth of all cell tested in a dose dependent manner. MPSP-1, MPSP-2 & MPSP-9 efficiently and MPSP-5 moderately inhibited the growth of both cell line tested with low IC50 value. They can still be considered good candidates for the development of anticancer drugs since they were shown to have activities against cancer cells. These compounds also can be used as drug leads for the development of other drugs or they can be used in combination with other antineoplastic drugs to compliment the therapeutic effects. However, more in- vitro and in-vivo mechanistic studies are required to understand the full potential of these compounds.

 

ACKNOWLEDGEMENTS:

The authors thank the IKG Punjab Technical University, Kapurthala, Jalandhar where the author is registered as a research scholar. The author also thanks Deshpande Laboratories Pvt. Ltd. Bhopal for evaluating the synthesized compounds for cytotoxic studies and sophisticated analytical instruments facility, Panjab University, Chandigarh for 1H-NMR, IR and Mass spectroscopic data.

 

CONFLICTS OF INTEREST:

There are no conflicts of interest.

 

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Received on 05.08.2020              Modified on 15.11.2020

Accepted on 01.02.2021             © RJPT All right reserved

Research J. Pharm. and Tech. 2021; 14(10):5101-5107.

DOI: 10.52711/0974-360X.2021.00889