INTRODUCTION:

Cancer is one of the most important and widespread health condition in the world, regardless of its type¹. It negatively impacts a patient’s quality of life and is one of the most common causes of death. Many drugs, treatments, and procedures have been discovered that can be used to treat cancer^2,3,4, such as radiotherapy⁵ which destroys the DNA of malignant cells, immunotherapy⁶andother therapeutic methods.

With the advancement of science and reaserch, there are now many targets that can play a significant role in the initation, progress, and even spread and metastasis of cancer.

One of these targets is carbonic anhydrase enzymes, as the tumor depends greatly on the PH intracellular and extracellular⁷, and the role of these enzymes in cancerous state is to provide the appropriate acidic environment for the oncogenic growth, and spread of malignant.

The role of carbonic anhydrase enzymes can be summarized by raising the PH intracellular to suit metabolic processes, and protecting cancer cells from death and also acidification of the extracellular milieu by decreasing PH value, which gives cancer cells an advantage and survival over normal cells that cannot live and adapt^8,9.

CA XII belongs to the carbonic anhydrase family (CAs), it catalyzes the hydration of carbon dioxide to Hco₃^- and H⁺^10,11. This process has a vital role in regulation of pH and CO₂ levels in multiple organs and tissues. It is found in plasma membrane of cells, especially in kidney tissue, epithelial cells, and CNS. CA XII is a membrane-bound enzyme, and it has a Zn⁺² in the active region, which is required for its activity. The expression and activity of this enzyme, in addition to the IX isozyme, increases in many cases, including hypoxia (low oxygen), which is the characteristic condition in tumors, so it’s an interesting target for therapeutic strategies in cancer ¹^2,¹^3,¹^4,15.

Sulfonylurea derivatives are commonly used to treat type 2 diabetes, but many other therapeutic uses have been discovered, such as diuretics, anti-inflammatory agent, antitubercular activity, antimalarial activity, anticancer, and others¹⁶.

Sulfonylurea can be used as CA XII inhibitors which target the active region of CA XII by binding to the amino acids in the active site and ionic bond forming with Zn⁺², then blockage of the bicarbonate obtaining process, and suppression of the catalyzing activity of CA XII^17,18.

Finally, there are several approved methods to synthesis sulfonylureas, and a new method has been developed in our lab, at the University of Aleppo, department of pharmaceutical chemistry with good yield and economical cost, in addition to being a single step, so it is easy to apply.

Many research in our laboratory at the University of Aleppo, Faculty of Pharmacy, has focused on the synthesis of various new sulfonamide derivatives with different pharmaceutical uses, including anti-bacterial, anti-cancer, and EGFR-inhibitory^{19,20,21,22,23}.

In this research, we have directed our efforts to design, develop and synthesize a novel sulfonylurea derivatives as potential anticancer and analyze them using many technical tools such as TLC, Ir spectro, mass spectro, and NMR to determine the structure of synthesized compounds.

Moreover, molecular modeling studies were done to determine the interaction of designed compounds with the target enzyme (carbonic anhydrase XII).

MATERIALS AND METHODS:

All of solvents and chemical materials were obtained from trusted source (Pro lab. For pharmaceutical industry). The enzyme code (PDB ID: 4WW8) has been downloaded from bank of protein data. The compounds were drawn and designed using the Marvin sketch program version 21.2 where the lowest energy conformer was selected, stored in molecular 2 format, and then applied to the Molegro Virtual Docker program (MVD) version 2022.5.5 to test their activity on the enzyme and to calculate the binding energy. The following devices were used to identify the compounds: Thin-layer separation method (tlc) with silica gel 60 F254 aluminum sheets (Macherey-Nagel Germany) to determine the Retardation Factor (RF) of materials, melting point of substance determined using BÜCHI Melting Point B-540 apparatus (BÜCHI Labortechnik, Switzerland), ATR-FTIR Bruker spectrophotometer (Bruker, Billerica, Massachusetts) to find out IR spectra, mass spectroscopy by using a mass spectrometer (Sciex, Framingham, USA), ¹H-NMR, ¹³C-NMR spectroscopy by using an NMR spectrometer (Joel, Tokyo, Japan), operating at 500 MHz for 1H and 13C.

Procedure of synthesis:

Synthesis of p-methoxybenzenesulfonyl chloride:

0.2mol of chlorosulfonic acid (13.3ml) was added to 0.1 mol anisole (10.9ml). Then, the content of reaction was stirred for 1hour in ice bath, then the crushed ice was added to the reaction to give p-methoxybenzenesulfonyl chloride precipitate. The resulting material was collected using vacuum filtration and it was stored in the refrigerator to prevent melting.

Synthesis of p-acetamidobenzenesulfonyl chloride:

0.5mol of chlorosulfonic acid (33.3ml) was added to 0.1 mol acetanilide (13.5g) in cold-bath. Then, the content of reaction was stirred for 2hour at 60C°. The precipitate of the substance is obtained by pouring ice to the reaction mixture.

Synthesis of compound A1:

0.01mol of 1-(4-flourophenyl) urea (1.54g) was dissolved in 10ml of acetone, 0.01mol of p-methoxybenzenesulfonyl chloride (2.06g) was added to the solution, 0.2ml of tri-ethyl-amine was added to the mixture of reaction with stirring at 60C⁰ under reflux condition for 3hours. Reaction mixture was poured onto ice to produce a precipitate.

Synthesis of compound B4:

0.01mol of urea (0.6g) was dissolved in 10ml of acetone, 0.01mol of p-acetamidobenzenesulfonyl chloride (2.34g) was added to the solution, 0.2ml of tri-ethyl-amine was added to the mixture of reaction with stirring at 60C⁰ under reflux condition for 3hours. Reaction mixture was poured onto ice to produce a precipitate.

Synthesis of compound D1:

0.01mol of 1-(4-flourophenyl) urea (1.54g) was dissolved in 10ml of acetone, 0.01mol of p-acetamidobenzenesulfonyl chloride (2.34g) was added to the solution, 0.2ml of tri-ethyl-amine was added to the mixture of reaction with stirring at 60C⁰ under reflux condition for 3hours, then the acetyl in the resulting substance is hydrolyzed using hydrochloric acid (4M) and heated in a water bath at a 80-90C⁰ for two hours. After that, the reaction mixture is cooled and the PH is adjusted using sodium bicarbonate. The crystals are waited for and then the recrystallized using water and ethanol (50:50).

A1 B4 D1

Carbonic anhydrase XII structure:

Carbonic anhydrase XII enzyme plays a critical role in regulating bicarbonate levels in tissues and the balance of PH but in tumors CAXII is overexpressed and it modulates PH levels in a way that helps facilitate invasion and spread of malignant by promoting the breakdown of the extracellular matrix and evading immune responses. Therefore, inhibiting it can greatly help prevent the development of cancer and even its spread to other tissues and organs²^4,²⁵.

The Protein data bank provided the Three-dimensional crystallized structure of carbonic anhydrase XII (pdb id:4ww8), which has a resolution of 1.42A⁰. It consists of four peptide chains A, B, C, D and 354 amino acids residues.

The crystallized structure includes 3 bound ligands: 4*ZN, 13*EDO, and 4*VD9²⁶.

Figure 1: Co-crystallized structure of the carbonic anhydrase XII (pdb:4ww8)

Docking studies:

Protein preparation:

The carbonic anhydrase XII protein was prepared by inserting it into the MVD program. A list of its amino acid residues is displayed, then any errors that may be present in the amino acid residues in the list are identified by colouring them in red or yellow. There are two types of possible errors that can occur in residues: deletion of atoms or errors in the bonds present in them. These faults are corrected to obtain the optimal structure of protein. In addition, Water molecules is removed from the protein’s crystal structure, the B chain was chosen to docking study, and the Zn⁺² ion in the active region was retained as cofactor, and everything else was removed.

Figure 2: Carbonic anhydrase XII cavity

Parameters used:

As shown in the table 1: the following parameters were used in docking studies:

Table 1: the used parameters for docking studies

Parameters	Value
Cavity volume	Cavity 1: 71.17 surface 212.48
Scoring function	Moldoc Score {GRID}
Grid resolution )Å(	0.30
Binding side radius	15
Searching-algorithm	Moldock optimizer
Number of runs	10
Max iteration	2000
Max population size	50
Energy threshold	100
Simplex evaluation max steps	300
Neighbour distance factor	1
Max number of poses returned	5

Validation docking method:

Molecular modeling was carried out to find out the possibility of compounds A1-E4 to inhibition of the carbonic anhydrase XII enzyme and to calculate the binding energies. But before that, the appropriate protocol must be determined and its validity verified. A method was developed in which the ligand(VD9:4-propylthiobenzenesulfonamide) bonded to the crystallized structure (PDb id 4ww8) was re-docked and then calculating of the standard deviation (Rmsd). The value for the used method was 1.41, which is considered an acceptable value, as in computer-aided drug design, standard deviation of less than 2 A⁰ is considered good.

This validated protocol was then used to perform molecular docking for the compounds A1-E4 where they were all compound in the same active pocket as shown in the Figure 3.

Figure 3: All compounds binding with CAXII binding pocket

RESULTS AND DISCUSSIONS:

Chemistry:

The synthesis of the compounds A1, B4, D1 was carried out in an alkaline medium of acetone and TEA, as the acidic medium leads to protonation of the terminal amino group in the phenyl urea derivatives, and the weakening of its nucleophiles,TEA abstracts a proton from the terminal NH2 group of the urea/phenyl urea derivatives to form a nucleophilic NH^-, then this nucleophilic NH^- attacks the electrophilic sulfur in SO₂Cl, displacing Cl^-, the Cl^- reacts with Et₃NH⁺ to form salt (Et₃NH⁺Cl^-), and acetone acts as a polar, non-protic solvent

Figure 4: synthesis of sulfonylurea derivatives

Docking:

The designed compounds and the reference compound(AZM) were drawn using the Marvin sketch program, converted to molecular 2 format, and then docking with Molegro Virtual Docker (MVD).

All compounds were bound to the CA XII enzyme and given a good binding energies. Also, all applied compounds formed hydrogen bounds with enzyme’s residues and ionic bounds with the Zn⁺² cofactor, with varied binding energies and bound distances. The binding-energies of all the compounds were higher than that of the native ligand and most of them had even higher binding-energies than the reference compound. Forming Hydrogen bound with Thr199 or Thr198 is essential for good binding energy with CA XII enzyme (pdb:4ww8) Forming more hydrogen bound with other enzyme’s residues gives an increase in binding energy, especially His93 and His91.

The compounds B1, B2, B3 have the highest binding energy with enzyme (158.20-162.85-153.74)Kcal/mol and this can be attributed to their formation of essential hydrogen-bonds with both Thr198, Thr199 and one of the amino acids which increases the energy His91 or His 93, and we note that the compounds A4, B4, C4, D4, E4 have the lowest binding energy among the studied compounds, and this may be due to the presence of a free urea group not attached to an aromatic ring, as the ring is formed steric interaction with many amino acids, which directly affects the energy. Compared to the standard compound, we note that the sulfonylurea group contributed significantly to increasing the binding energy by forming additional bonds with amino acids through S=O, NH, C=O groups and, of course, through the steric interactions it possesses.

Figure 5.6: Predicted binding positions of compound A1 and the interactions with amino acids residues in the binding poket of CAXII.

Table 2. Binding energies (Kcal/mol) of the studied compounds compared to the standard ligand (-113.46) Kcal/mol and the reference compound acetazolamide (-120.97) Kcal/mol.

compound	Binding energies (Kcal/mol)	compound	Binding energies (Kcal/mol)	compound	Binding energies (Kcal/mol)
A1	-146.75	B4	-132.67	D3	-125.38
A2	-131.97	C1	-143.93	D4	-118.55
A3	-148.29	C2	-140.95	E1	-149.79
A4	-119.12	C3	-138.63	E2	-149.48
B1	-158.20	C4	-118.32	E3	-146.97
B2	-162.85	D1	-144.36	E4	-115.60
B3	-153.74	D2	-126.14

Table 3. The interaction of studied compounds with amino acids of CA XII enzyme:

Ligand	Residue	Hydrogen bond	Distance (A⁰)	Energy	Steric interaction
Reference AZM	Thr199	OH…SO(Ligand) NH…SO(Ligand)	2.70 3.22	-1.24 -1.88	-
	Thr198	OH…NH(Ligand)	3.35	-1.25
	His93	N(imidazole)…NH(Ligand)	3.19	-2.05
	His91	N(imidazole)…NH(Ligand)	2.85	-2.50
Standard	Thr199	OH…SO(Ligand) NH…SO(Ligand)	3.11 2.72	-2.47 -0.97	Thr198
	Thr198	OH…NH(Ligand)	2.94	-2.50
	His93	N(imidazole)…NH(Ligand)	2.96	-2.50
	His91	N(imidazole)…NH(Ligand)	3.05	-2.50
A1	Thr199	NH…CO(Ligand) OH…NH(Ligand)	2.95 3.13 2.67 2.74	-0.69 -1.06 -2.50 -1.53	Gln89, Ser67, Asn64,Val141, His117, Leu197, Val119, His91
	Thr198	OH…CO(Ligand) NH…CO(Ligand)
A2	Thr198	OH…CO(Ligand)	2.56 2.75 2.98	-2.13 -2.50 -2.50	Leu197, His91, Ser67, His93,Thr199
	Tyr6	OH…SO(Ligand)
	Ser67	OH…SO(Ligand)
A3	Thr198	OH…CO(Ligand) NH…CO(Ligand) NH…SO(Ligand)	2.83 3.28 3.15 3.26	-2.50 -0.88 -2.17 -1.68	His66, His91, Val141, Trp208, His117, Leu197, Val119,Thr199, His93
	Thr199	OH…CO(Ligand)
A4	Thr198	NH…CO(Ligand) OH…CO(Ligand) OH…SO(Ligand) NH…SO(Ligand)	2.97 3.25 3.14 3.00 2.97 2.76	-2.29 -1.74 -2.30 -1.20 -2.50 -0.73	Trp208, His91, Thr199
	Thr199	OH…SO(Ligand) NH…SO(Ligand)
B1	Thr198	OH…CO(Ligand) NH…SO(Ligand)	2.69 2.71 3.25 2.99	-2.50 -2.50 -1.76 -1.82	Asn64, His66, His93, His117, Val119, Ser133, Thr199
	Thr199	OH…CO(Ligand)
	His91	N(imidazole)…NH(Ligand)
B2	Thr198	NH…SO(Ligand) OH…SO(Ligand) OH…CO(Ligand)	2.75 3.19 2.64 3.36 3.06	-2.50 -2.03 -2.50 -1.22 -1.38	Asn64, His117, His93, Thr199
	Thr199	OH…CO(Ligand)
	His91	N(imidazole)…NH(Ligand)
B3	Thr198	NH…SO(Ligand) NH…CO(Ligand) OH…CO(Ligand)	3.01 3.33 2.54 3.40 2.97 2.74	-2.50 -0.65 -1.65 -1.02 -2.39 -2.50	Asn64, His91, His93, His117, Thr198, Thr199
	Thr199	OH…CO(Ligand)
	His93	N(imidazole)…NH(Ligand)
	Gln89	NH…CO(Ligand)
B4	Thr198	OH…CO(Ligand) NH…CO(Ligand) OH…SO(Ligand) NH…SO(Ligand)	3.25 2.96 3.14 2.97 2.97 2.75	-1.73 -2.30 -2.28 -1.20 -2.50 -0.76	His91, Trp208, Thr199
	Thr199	OH…SO(Ligand) NH…SO(Ligand)
C1	Thr198	OH…CO(Ligand) NH…CO(Ligand) OH…SO(Ligand)	2.95 3.08 2.47 2.78	-2.50 -2.50 -1.41 -2.50	Leu197, Val119, Thr199, Asn64, His91
	Thr199	OH…SO(Ligand)
C2	Thr198	OH…CO(Ligand) NH…CO(Ligand) NH…SO(Ligand)	2.95 3.35 2.96 3.15 2.97	-1.52 -0.66 -2.44 -2.26 -2.43	His66, His117, Trp208, Leu197, Val119, His93, Thr199
	Thr199	OH…CO(Ligand)
	His91	N(imidazole)…NH(Ligand)
C3	Thr198	OH…CO(Ligand) NH…CO(Ligand) OH…SO(Ligand)	3.02 3.02 2.39 2.83	-2.50 -2.48 -0.79 -2.50	Leu197, Val119, Thr199, Asn64, His91
	Thr199	OH…SO(Ligand)
C4	Thr198	OH…CO(Ligand) NH…CO(Ligand)	3.05 3.08 2.98	-2.50 -2.47 -2.27	His91, Thr199, Ser67
	Thr199	OH…SO(Ligand)
D1	Thr198	OH…SO(Ligand) NH…SO(Ligand)	3.26 2.76 3.16	-1.71 -2.50 -1.66	Ser67, Asn64, Gln89, Thr199, His117, Trp208, Thr198, Leu197, Val119, His91
	Thr199	OH…NH(Ligand)
D2	Thr198	OH…CO(Ligand)	2.52 2.83 3.01	-1.19 -1.60 -1.06	Thr198,Thr199, His91, His93, Ser67
	Tyr6	OH…SO(Ligand)
	Ser67	OH…SO(Ligand)
D3	Thr198	OH…CO(Ligand)	2.80 3.24 2.72 2.97	-2.00 -1.43 -1.40 -1.09	Val141, Trp208, His91, Ser67, Asn64, His93, Thr199
	Thr199	OH…CO(Ligand)
	Tyr6	OH…SO(Ligand)
	Ser67	OH…SO(Ligand)
D4	Thr198	NH…CO(Ligand) OH…SO(Ligand) NH…SO(Ligand)	3.10 3.12 2.95 3.01 2.76	-2.26 -2.42 -1.22 -2.50 -0.76	Trp208, Thr199, His93, Asn64
	Thr199	OH…SO(Ligand) NH…SO(Ligand)
E1	Thr198	OH…SO(Ligand) NH…SO(Ligand) OH…CO(Ligand)	3.28 2.68 2.58 3.46 3.07	-1.60 -2.50 -2.37 -0.68 -1.45	Thr199, His66, His93
	Thr199	OH…CO(Ligand)
	His91	N(imidazole)…NH(Ligand)
E2	Thr198	OH…CO(Ligand) NH…CO(Ligand) NH…SO(Ligand)	2.88 3.32 3.00 3.21 2.98	-2.50 -0.77 -2.45 -1.92 -2.19	His66, His117, Val141, Trp208, Leu197, Val119, Thr199
	Thr199	OH…CO(Ligand)
	His91	N(imidazole)…NH(Ligand)
E3	Thr198	OH…CO(Ligand) NH…CO(Ligand) NH…SO(Ligand)	3.00 3.34 3.02 3.13	-2.50 -0.70 -2.39 -2.35	Thr199, His93, His66, Val141, Trp208, Val119, Leu197, His91, His117
	Thr199	OH…CO(Ligand)
E4	Thr198	NH…CO(Ligand) OH…SO(Ligand) NH…SO(Ligand)	2.97 3.16 3.00 2.95 2.75	-2.29 -2.21 -1.19 -2.50 -0.75	Trp208, Thr199, His91
	Thr199	OH…SO(Ligand) NH…SO(Ligand)

Analytical Data:

Compound A1:

1‐(4‐fluorophenyl) ‐3‐(4‐methoxybenzenesulfonyl) urea

White powder, yield 87.5%, melting point 182-186C⁰, tlc: the solvent system was ethyl acetate: petrolatum ether 2.5:7.5, and the retention factor for compound A1 is 0.32. Ir spectro (vmax, cm^-1): 3417 (N-H), 3052 (C-H aroma), 1654(C=O), 1410(C-H), 1357(S=O), 1230(C-O).1H NMR spectro (DMSO-d6, δ, ppm) δ 9.80 (s, 1H, N-H), 7.85 (d, 2H, Ar-H), 7.47 (dd, 2H Ar-H), 7.18 (d, 2H, Ar-H), 7.10 (t, 2H, Ar-H), 6.40 (s, 1H, N-H), 3.73 (s, 1H, C-H). 13C-NMRspectro (DMSO-d6, δ, ppm): 162.11, 157.97, 156.29, 137.29, 127.47, 126.61, 132.66 – 106.55, 119.82, 118.37, 115.53, 56.91. Mass spectrum (m/z, ESI): showed molecular ion (M+H) ⁺ peak at 325.15 representing C₁₄H₁₄FN₂O₄S. and (M+Na) peak at 347.22, and its fragments are 154.27, 172.19, 186.85.

Compound B4:

N‐{4‐[(carbamoylamino)sulfonyl] phenyl} acetamide

White powder, yield 84.5%, melting point 212-216 C⁰, tlc: the solvent system was ethyl acetate: petrolatum ether 8.0:2.0, and the retention factor for compound B4 is 0.32. Ir spectro (vmax, cm^-1): 3416 (N–H), 3050 (C–H aroma), 1662 (C=O), 1402 (C-H), 1322 (S=O). 1H NMR spectro (DMSO-d6, ppm) 10.31 (s, 1H, N-H), 7.77 (d, 2H, Ar-H), 7.62 (d, 2H, Ar-H), 6.60 (s, 2H, NH2), 5.97 (s, 1H, N-H), 2.06 (s, 3H, C-H). 13C-NMR spectro (DMSO-d6, δ, ppm): 169.56, 155.65, 143.82, 129.14, 118.96, 39.77, 25.20. Mass spectrum (m/z, ESI): showed molecular ion (M+H) ⁺ peak at 258.18 representing C₉H₁₁N₃O₄S. and (M+Na) peak at 280.09, and its fragments are 198.21, 122.05.

Compound D1:

3‐(4‐aminobenzenesulfonyl) ‐1‐(4‐fluorophenyl) urea Light violet crystals, yield 82.0%, melting point 193-197 C⁰, tlc: the solvent system was ethyl acetate: petrolatum ether 8.0:2.0, and the retention factor for compound D1 is 0.43. Ir spectro (vmax, cm^-1): 3413(N–H), 1635(C=O urea), 1335 (S=O).1H NMR spectro (DMSO-d6, δ, ppm) δ 8.58 (s, 1H, N-H), 8.01 (d, 2H, Ar-H), 7.57 – 7.43 (m, 2H, Ar-H), 7.36 (dd, 2H, Ar-H), 7.07 – 6.93 (m, 2H, Ar-H), 6.19 (s, 2H, NH2), 5.79 (s, 1H, N-H).13C-NMR spectro (DMSO-d6, δ, ppm): 158.35, 156.57, 137.29, 127.47, 126.61, 119.77, 115.60, 115.43. Mass spectrum (m/z, ESI) showed molecular ion (M+H) ⁺ peak at 310.25 representing C₁₃H₁₂FN₃O₃S. and (M+Na) peak at 332.18, and its fragments 155.10, 177.10.

CONCLUSION:

The above article describes the characterization and synthesis of new sulfonylurea derivatives as anticancer drugs by potentially targeting the carbonic anhydrase XII enzyme. New sulfonylurea derivatives were synthesized in good yields and by a simple and easy method by reacting (sulfonyl chloride) derivatives with phenyl urea derivatives in a medium of acetone and TEA.

The structures of these compounds were identified using 13C and 1H NMR, IR, and Mass spectro. The efficiency of the designed and synthesized compounds was determined by docking them into the CA XII (pdb:4ww8) binding cavity using MVD program (Molegro Virtual Docker). The binding energies of these compounds were calculated and compared with the energy of the reference ligand.

Based on these results, we found that all of the designed and synthesized compounds have higher binding-energies than the native ligand and some of them have higher binding-energies than acetazolamide due to the formation of hydrogen-bonds and additional steric interactions with the amino acids within the enzyme.

Finally, there is of course a need for more research to evaluate the effectiveness of these compounds as anti-cancer drugs by inhibiting the CA XII enzyme, and we hope that this study will give the desired effectiveness and promising results.

ACKNOWLEDGMENTS:

The authors of this article thank the Department of Pharmaceutical Chemistry, Faculty of pharmacy, university of Philipps, Marburg, Germany, for the analysis of the synthesized compounds using 13C and 1H NMR, and Mass spectroscopy.

CONFLICTS OF INTEREST:

The authors of the article declare no conflicts of interest regarding the publication of this paper.

REFERENCES:

1. Kumbhar Swapnil, Salunkhe Vijay , Magdum Chandrakant.Targeted Drug Delivery: A Backbone for Cancer Therapy. Asian J. Pharm. Res. 2013; 3(1): 40-46.

2. Baldev Krishan Sharma. Synthetic and Natural Compounds as Anti-Cancer Agents – A Review . Asian J. Research Chem. 2017; 10(5): 699-707.

3. Ganesh G. Dhakad, Gayatri D. Patil, Ashwini C. Nikum, Sangita P. Shirsat. Review on Radiation Therapy on Cancer. Research Journal of Pharmacology and Pharmacodynamics. 2022; 14(1): 4-2

4. T. Angel Priya, N. Ganapathy, P. Padmavathi. Guided Imagery on Bio Physiological Parameters among Patients receiving Chemotherapy. Asian J. Nursing Education and Research. 2018; 8(3): 437-439

5. Sonja Jose, B. R. Balakrishnan. Cervical Cancer Prevention and Treatment: An Overview. Research Journal of Pharmacy and Technology. 2021; 14(4): 2353-9

6. GK Sudhakar, Vasudev Pai, Arvind Pai. An overview on current Strategies in Breast Cancer Therapy. Research J. Pharmacology and Pharmacodynamics. 2013; 5(6): 353-355 .

7. Gerweck LE. Tumor pH: Implications for treatment and novel drug design. Semin Radiat Oncol. 1998; 8(3): 176-182. doi:10.1016/S1053-4296(98)80020-5

8. Mboge MY, Mahon BP, McKenna R, Frost SC. Carbonic anhydrases: role in pH control and cancer. Metabolites. 2018; 8(1): 19. doi:10.3390/metabo8010019

9. Supuran CT. How many carbonic anhydrase inhibition mechanisms exist? J Enzyme Inhib Med Chem. 2016; 31(3): 345-360. doi:10.3109/14756366.2015.1122001

10. Gupta A, Lather V, Vennapu DR, Kumar S, Dhiman N, Sharma A. 3D-QSAR, E-pharmacophore and molecular docking to explore substituted sulfonamides as carbonic anhydrase inhibitors in epilepsy. Res J Pharm Technol. 2022; 15(9): 4157-4165. doi:10.52711/0974-360X.2022.00932

11. Das P, Nayak A, Bhavani K, Deepika, Ranjitha, Md Afsar. Molecular modelling technique on interactivity between human carbonic anhydrase 1 and mangiferin for antiulcer activity. Res J Pharm Technol. 2023; 16(5): 2219-2224. doi:10.52711/0974-360X.2023.00406

12. Pastorekova S, Gillies RJ. The role of carbonic anhydrase IX in cancer development: links to hypoxia, acidosis, and beyond. Cancer Metastasis Rev. 2019; 38(1-2): 65-77. doi:10.1007/s10555-019-09799-0

13. Hassan MI, Shajee B, Waheed A, Ahmad F, Sly WS. Structure, function and applications of carbonic anhydrase isozymes. Bioorg Med Chem. 2013; 21(6): 1570-1582. doi:10.1016/j.bmc.2012.11.051

14. Kopecka J, Campia I, Jacobs A, et al. Carbonic anhydrase XII is a new therapeutic target to overcome chemoresistance in cancer cells. Oncotarget. 2015; 6: 6776-6793. doi:10.18632/oncotarget.2882

15. Rollando Rollando, Warsito Warsito, Masruri Masruri, Nashi Widodo. Potential matrix metalloproteinase-9 inhibitor of aurone compound isolated from Sterculia quadrifida leaves: In-vitro and in-silico studies. Research Journal of Pharmacy and Technology. 2022; 15(11): 5250-4.

16. Sen S, Kumar DR, Ruchika, Easwari TS, Gohri S. Therapeutic aspects of sulfonylureas: a brief review. J Chem Pharm Res. 2016; 8(12): 121-130.

17. Idrees D, Hadianawala M, Mahapatra AD, et al. Implication of sulfonylurea derivatives as prospective inhibitors of human carbonic anhydrase II. Int J Biol Macromol. 2018; 115: 961-969. doi:10.1016/j.ijbiomac.2018.04.038

18. Hadianawala M, Datta B. Design and development of sulfonylurea derivatives as zinc metalloenzyme modulators. RSC Adv. 2016; 6: 8923-8929. doi:10.1039/C5RA27341B

19. Mohamad M, Djamila BH, Amir B, Mustapha FC. Molecular Modeling Study of Bis-sulfonamide Derivatives Synthesis Targeting Aromatase Enzyme as Anticancer. Research Journal of Pharmacy and Technology. 2024; 17(1): 43-0. doi: 10.52711/0974-360X.2024.00008.

20. Mohammad Hasbi Ghazal, Djamila Ben Hadda, Saleh Trefi, Amir Balash, Mustapha Fawaz Chehna. New Synthetic Methods of Ureido-Substituted Benzenesulfonamides as Carbonic Anhydrase IX Inhibitors. Research Journal of Pharmacy and Technology. 2023; 16(8): 3884-0. doi: 10.52711/0974-360X.2023.00640.

21. Akili, S. , Ben Hadda, D. , Bitar, Y. , Najjar, A. and Fawaz Chehna, M. (2021) Computer-Aid Design of Novel Sulfonamide Derivatives as EGFR Kinase Inhibitors for Cancer Treatment. International Journal of Organic Chemistry, 11, 171-186. doi: 10.4236/ijoc.2021.114012.

22. Hussein MH, Ibrahim A, Anan MT, Chehna MF. Synthesis of aryl sulfonamides by eco-friendly method and evaluation of their anti-bacterial activity. Int J Chem Biochem Sci. 2023; 23(1): 363-371.

23. Kattan RMR, Ben Hadda D, Balash A, Chehna MF. Design, facile synthesis, and characterization of sulfonamide derivatives combined with amino acids and other drugs targeting HER2. Res J Pharm Technol. 2024; 17(2): 264. doi:10.52711/0974-360X.2024.00264

24. Waheed A, Sly WS. Carbonic anhydrase XII functions in health and disease. Gene. 2017; 623: 33-40. doi:10.1016/j.gene.2017.04.066

25. Daunys S, Petrikaitė V. The roles of carbonic anhydrases IX and XII in cancer cell adhesion, migration, invasion and metastasis. Biol Cell. 2020; 112(12): 383-397. doi:10.1111/boc.201900099

26. Smirnov A, Manakova E, Grazulis S. Crystal structure of human carbonic anhydrase isozyme XII with 4-propylthiobenzenesulfonamide. RCSB Protein Data Bank. Published 2015. Accessed April 14, 2025. doi:10.2210/pdb4WW8/pdb

Received on 28.04.2025 Revised on 18.08.2025

Accepted on 24.11.2025 Published on 20.05.2026

Available online from May 25, 2026

Research J. Pharmacy and Technology. 2026;19(5):2075-2081.

DOI: 10.52711/0974-360X.2026.00298

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. Creative Commons License.