|Year : 2004 | Volume
| Issue : 4 | Page : 234-237
Cardio-respiratory effects of DRDE-07, a new prophylactic agent for sulphur mustard toxicity, in anesthetized rats
V Malviya , R Singh , D Kumar , U Pathak , P Kumar , DK Jaiswal , R Mathur , R Vijayaraghavan
Defence Research and Development Establishment. Jhansi Road, Gwalior - 474 002, India
|Date of Submission||12-Jul-2003|
|Date of Decision||08-Apr-2004|
|Date of Acceptance||09-Apr-2004|
Defence Research and Development Establishment. Jhansi Road, Gwalior - 474 002, India
OBJECTIVE: To study the cardio-respiratory effects of DRDE-07, a new prophylactic agent for sulphur mustard.
MATERIAL AND METHODS: The effect of DRDE-07 was studied in anesthetized male rats (Wistar), by administering it either by intraperitoneal or oral routes. Different doses (0.25, 0.5, 1.0 and 2.0 LD50) were used and various cardio-respiratory parameters viz., mean arterial blood pressure, heart rate, respiratory rate and tidal volume were monitored on an oscillograph.
RESULTS: Intraperitoneal administration of 0.5 and 1.0 LD50 and oral administration of 0.25 LD50 of DRDE-07 did not produce any effect on the cardio-respiratory variables. Oral administration of 0.5 and 1.0 LD50 showed a significant decrease in mean arterial blood pressure after 60 min. None of the animals died in the two-hour monitoring period. Two (2.0) LD50 administration of DRDE-07 either by intraperitoneal or oral routes induced a sudden decrease in the mean arterial blood pressure and the animals died within one hour.
CONCLUSION: DRDE-07 did not show any significant effect on the cardio-respiratory variables at a dose of 0.25 LD50 by oral route and 1.0 LD50 by intraperitoneal route. The death after intraperitoneal or oral administration (2.0 LD50) of DRDE-07 is due to the sudden fall in the mean arterial blood pressure.
|How to cite this article:|
Malviya V, Singh R, Kumar D, Pathak U, Kumar P, Jaiswal D K, Mathur R, Vijayaraghavan R. Cardio-respiratory effects of DRDE-07, a new prophylactic agent for sulphur mustard toxicity, in anesthetized rats. Indian J Pharmacol 2004;36:234-7
|How to cite this URL:|
Malviya V, Singh R, Kumar D, Pathak U, Kumar P, Jaiswal D K, Mathur R, Vijayaraghavan R. Cardio-respiratory effects of DRDE-07, a new prophylactic agent for sulphur mustard toxicity, in anesthetized rats. Indian J Pharmacol [serial online] 2004 [cited 2020 Feb 29];36:234-7. Available from: http://www.ijp-online.com/text.asp?2004/36/4/234/11149
| » Introduction|| |
The Chemical Weapons Convention (CWC) came into force from 29 April 1997, prohibiting the production, storage, transport and use of chemicals on enemy forces. Countries which had already stockpiled the chemical weapons were given a time of 10 years to destroy them. One such chemical is sulphur mustard (SM), commonly known as mustard gas. Reports are available of its use in several instances.,, Huge quantities of the stockpiled SM are being destroyed by several countries. In spite of the CWC, SM can be used clandestinely during war or by terrorists due to its simple method of preparation.
SM, chemically bis [2-chloroethyl] sulphide, is an alkylating agent that causes serious blisters upon contact with human skin. In animal models it is extremely lethal. Just one microliter applied on the skin of a mouse or a rat may be lethal in a weeks time. SM forms sulphonium ion in the body and alkylates DNA leading to DNA strand breaks and cell death., Due to the high electrophilic property of the sulphonium ion, SM binds to a variety of cellular macromolecules and fatality may occur due to multi-organ failure., The eyes, skin and the respiratory tract are the principal target organs of SM toxicity., , 
SM is highly lipophilic and is absorbed very quickly after contact with the skin. Though several antidotes have been reported for the systemic toxicity of SM in experimental animals,,,,, none of them have been approved so far and decontamination of SM immediately after contact is recommended as the best protection. Hence, the decontamination has to be achieved quickly to limit the absorption. The most commonly used decontaminant is Fuller's earth (a native form of aluminum silicate) that removes SM by adsorption, thereby reducing the toxicity. A few chemical decontaminants for human use have also shown very good efficacy.,
An effective prophylactic agent against SM toxicity is the requirement of the day, especially for personnel engaged in the destruction of SM and during inspection by the Organization for Prohibition of Chemical Weapons. From a series of aminothiols, two compounds amifostine [organophos-phorothioate] and DRDE-07 [S-(w-aminoalkylamino) alkylaryl sulphide] gave very good protection as prophylactic agents against SM.,, When given orally DRDE-07 was more efficacious than amifostine as a prophylactic agent against SM induced percutaneous toxicity., The acute toxicity studies (LD50) of DRDE-07 have also been reported.
As DRDE-07 is an investigational drug, to introduce it as a prophylactic agent to the armed forces, various pharmacological and toxicological studies are needed. We initiated several of them and here we report its effect on cardio-respiratory variables.
| » Material and methods|| |
Chemicals: DRDE-07 was synthesized in the chemistry laboratory. The compound was characterized by elemental analysis, IR, 1H NMR and mass spectral analysis. The purity was checked by thin-layer chromatography. All other chemicals used were of analytical grade.
DRDE-07: NH2 - CH2 - CH2 - NH - CH2 - CH2 - S - C6H5
Animals : Wistar male rats (225-300 g, body weight), randomly bred and maintained in the animal house were used for the study. Thirty-six rats were used in the present study and they were divided into nine groups of four animals each. The animals were housed in polypropylene cages on dust-free rice husk as the bedding material. The animals were provided with pellet diet (Amrut Ltd., India) and water ad libitum. The care and maintenance of the animals were as per the approved guidelines of the Committee for the Purpose of Control and Supervision of Experiments on Animals, India. This study has the approval of the Establishment's Ethics Committee.
Cardio-respiratory parameters in anesthetized rats: The mean arterial blood pressure, heart rate, respiratory rate and tidal volume were recorded using a six-channel polygraph (Graphtec Linerecorder, Germany). The rats were anesthetized with urethane (1.6 g.kg-1, i.p.) and the neck region was dissected and exposed. The trachea was cannulated and connected to a pneumotachograph. The right jugular vein was cannulated and a syringe was connected for intravenous administrations. The left carotid artery was cannulated with a thin polypropylene tube connected to a pressure transducer (Statham P23Dc), filled with heparinised normal saline. The pressure transducer was connected to a preamplifier (low-level DC, Grass Instruments, USA) and arterial blood pressure was recorded on the polygraph. Mean arterial pressure was calculated from the recorded blood pressure. The signals from the DC Driver amplifier recording the blood pressure were taken from J6 output and fed into the external triggering (High) input of EKG Tachograph preamplifier (Grass Instruments, USA) for recording the heart rate. The pneumotachograph was connected to a differential pressure transducer (SWEMA, Germany) and connected to a pre-amplifier (Low level DC, Grass Instruments, USA), and inspiration was recorded as an upward deflection and expiration as a downward deflection. A polythene tube was inserted through the esophagus to the stomach for feeding DRDE-07. After the surgical procedures, the rat was allowed to stabilize for about 1 h. Three doses of DRDE-07 viz., 0.5, 1.0 and 2.0 LD50 (in distilled water) were administered intraperitoneally (LD50 = 141 mg/kg). Four doses of DRDE-07 viz., 0.25, 0.5, 1.0 and 2.0 LD50 (in distilled water) were administered orally (LD50 = 1599 mg/kg). The control group was administered distilled water.
All values were converted as percent of control and represented as mean+SEM. The data were analyzed by one-way ANOVA followed by Student-Newmann-Keuls multiple comparison procedure. P values <0.05 were considered statistically significant. SigmaStat (Jandel Sci., USA) was used for the statistical calculations.
| » Results|| |
The control values (mean+SE, n=4) of the mean arterial blood pressure, heart rate, respiratory rate and tidal volume in the anesthetized rats were 110+4 mmHg, 338+11 per minute, 37.0+3.3 per minute and 0.66+0.03 ml/min respectively. During the two-hour monitoring period the control animals showed a steady response.
Intraperitoneal administration of DRDE-07 did not show any effect on the blood pressure, heart rate, respiratory rate and tidal volume at 0.5 and 1.0 LD50 doses. At 2.0 LD50 dose the animals died within 60 minutes and a significant decrease in blood pressure and tidal volume was observed [Table - 1]. Oral administration of 0.25 LD50 of DRDE-07 did not show any significant change on the cardio-respiratory variables. 0.5 and 1.0 LD50 of DRDE-07 administered orally showed a significant decrease in the mean arterial blood pressure after 60 min. There was no change in the heart rate, respiratory rate and tidal volume. None of the animals died in the two-hour monitoring period following 1.0 LD50 dose. Oral administration of 2.0 LD50 of DRDE-07 induced a sudden decrease in the mean arterial blood pressure and the animals died within 60 min [Table - 2].
| » Discussion|| |
Antidotes to SM can act by four different mechanisms: (a) prevention of SM from entering the system (personal decontamination at the site of contact), (b) prevention of SM from alkylating critical target molecules mainly DNA, (c) retrieval of SM alkylated DNA and (d) prevention and reversal of the cascade of secondary biochemical reactions of alkylation. SM is known to react with a variety of macromolecules viz., DNA, proteins and several thiol-containing molecules including GSH. A decrease in the GSH content was reported following percutaneous administration of SM. If thiol-containing compounds are administered as prophylactic agents they can spare GSH by neutralizing SM. Amifostine is one such thiol-containing compound recommended as an adjunct to anticancer agents viz., cisplatin and cyclophosphamide which selectively protects normal tissues without reducing the effect of the anticancer agents on the cancer cells.,,,, Analogues of amifostine with better lipophilicity were synthesized that were expected to have better efficacy. Among the several compounds synthesized, DRDE-07 gave better protection when it was administered through the oral route, against percutaneously administered SM.,
Amifostine and DRDE-07 did not protect when SM was administered by oral, subcutaneous and inhalation routes. When SM was administered through percutaneous route amifostine and DRDE-07 were effective, showing that the mechanism of the toxic effect of SM varies with different routes. Interestingly, percutaneously administered SM was more toxic in the rodent species viz., mice and rats.
The cardio-respiratory effects of DRDE-07 were studied at various doses for two reasons: (i) to evaluate the safety of DRDE-07 and (ii) to ascertain whether DRDE-07 causes a profound fall in blood pressure that reduces the absorption of SM when the latter is administered through the percutaneous route. The LD50 of DRDE-07 by intraperitoneal and oral routes in male rats were 141 and 1599 mg/kg respectively. The doses that were employed for the prophylactic effect were 0.05, 0.1 and 0.2 LD50 doses, orally., In the present study up to 0.5 and 1 LD50 of DRDE-07 by the intraperitoneal route did not produce any change in the cardio-respiratory parameters. Oral administration of 0.25 LD50 did not show any effect on the blood pressure. So the possibility of the reduced absorption of SM when it is administered through the percutaneous route is ruled out at lower doses of DRDE-07.
In conclusion, DRDE-07 did not show any significant effect on the cardio-respiratory variables at a dose of 0.25 LD50 by the oral route and 1.0 LD50 by the intraperitoneal route. The death after intraperitoneal or oral administration (2.0 LD50) of DRDE-07 is due to a sudden fall in the mean arterial blood pressure.
| » Acknowledgement|| |
The authors are grateful to Mr. K. Sekhar, Director, DRDE, Gwalior for his constant encouragement.
| » References|| |
|1.||Krutzsch W, Trapp R. A commentary on the chemical weapons convention. London: Martinus Nijhoff Publishers; 1994. |
|2.||Smith WJ, Dunn MA. Medical defense against blistering chemical warfare agents. Archive Dermatol 1991;127:1207-13. [PUBMED] |
|3.||Eisenmenger W, Drasch G, von Clarmann M, Kretschmer E, Roider G. Clinical and morphological findings on mustard gas [bis(2-chloroethyl) sulfide] poisoning. J Forensic Sci 1991;36:1688-98. [PUBMED] |
|4.||Momeni AZ, Enshaeih S, Meghdadi M, Amindjavaheri M. Skin manifestations of mustard gas. A clinical study of 535 patients exposed to mustard gas. Arch Dermatol 1992;128:775-80. [PUBMED] |
|5.||Papirmeister B, Feister AJ, Robinson SI, Ford RD. Medical defense against mustard gas: Toxic mechanisms and pharmacological implications. Boca Raton: CRC Press; 1991. |
|6.||Lakshmana Rao PV, Vijayaraghavan R, Bhaskar AS. Sulphur mustard-induced DNA damage in mice after dermal and inhalation exposure. Toxicology 1999; 139:39-51. [PUBMED] |
|7.||Somani SM, Babu SR. Toxicodynamics of sulfur mustard. Int J Clin Pharmacol Ther Toxicol 1989;27:419-35. [PUBMED] |
|8.||Dacre JC, Goldman M. Toxicology and pharmacology of the chemical warfare agent sulfur mustard. Pharmacol Rev 1996;48:289-326. [PUBMED] |
|9.||Pechura CM, Rall DP. Veterans at risk: The health effects of mustard gas and lewisite. Washington DC: National Academy Press; 1993. |
|10.||Vijayaraghavan R. Modifications of breathing pattern induced by inhaled sulphur mustard in mice. Arch Toxicol 1997;71:157-64. [PUBMED] [FULLTEXT]|
|11.||Callaway S, Pearce KA. Protection against systemic poisoning by mustard gas, di (2-chloroethyl) sulphide, by sodium thiosulphate and thiocit in albino rat. Br J Pharmcol 1958;13:395-8. [PUBMED] |
|12.||Vojvodic V, Milosavljevic Z, Boskovic B, Bojanic N. The protective effect of different drugs in rats poisoned by sulfur and nitrogen mustards. Fundam Appl Toxicol 1985;5:160-8. |
|13.||Vijayaraghavan R, Sugendran K, Pant SC, Husain K, Malhotra RC. Dermal intoxication of mice with bis (2-chloroethyl) sulphide and the protective effect of flavonoids. Toxicology 1991;69:35-42. [PUBMED] |
|14.||Kumar O, Sugendran K, Vijayaraghavan R. Protective effect of various antioxidants on the toxicity of sulphur mustard administered to mice by inhalation or percutaneous routes. Chem Biol Interact 2001; 134: 1 - 12. |
|15.||Marrs TC, Maynard RL, Sidell FR. Chemical warfare agents; Toxicology and treatment. Chichester: John Wiley and Sons; 1996. |
|16.||Shih ML, Korte WD, Smith JR, Szafraniec LL. Reactions of sulfides with S-330, a potential decontaminant of sulfur mustard in formulations. J Appl Toxicol 1999;19 (Suppl 1):83-8. |
|17.||Vijayaraghavan R, Kumar P, Dubey DK, Singh R. Evaluation of CC2 as a decontaminant in various hydrophilic and lipophilic formulations against sulphur mustard. Biomed Environ Sci 2002;15:25-35. [PUBMED] |
|18.||Joshi U, Raza SK, Kumar P, Vijayaraghavan, R., Jaiswal, D.K. A process for preparation of S-(T-aminoalkylamino) alkylaryl sulphide dihydrochloride. New Delhi, India: Indian patent filed, Patent Office; 1999. |
|19.||Vijayaraghavan R, Kumar P, Joshi U, Raza SK, Lakshmana Rao PV, Malhotra RC, et al. Prophylactic efficacy of amifostine and its analogues against sulphur mustard toxicity. Toxicology 2001;163:83-91. [PUBMED] [FULLTEXT]|
|20.||Bhattacharya R, Rao PV, Pant SC, Kumar P, Tulsawani RK, Pathak U, et al. (2001). Protective effects of amifostine and its analogues on sulfur mustard toxicity in vitro and in vivo. Toxicol Appl Pharmacol 2001;176:24-33. [PUBMED] [FULLTEXT]|
|21.||Kumar P, Vijayaraghavan R, Kulkarni AS, Pathak U, Raza SK, Jaiswal DK. In vivo protection by amifostine and DRDE-07 against sulphur mustard toxicity. Hum Exp Toxicol 2002; 21: 371 - 6. |
|22.||Kumar P, Vijayaraghavan R, Singh M. Efficacy of atropine nasal aerosol spray against organophosphorus poisoning. Indian J Pharmacol 2001;33:431-4. |
|23.||Foster-Nora JA, Siden R. Amifostine for protection from antineoplastic drug toxicity. Am J Health Syst Pharm 1999;54:787-800. |
|24.||Hospers GA, Eisenhauer EA, de Vries EG. The sulfhydryl containing compounds WR-2721 and glutathione as radio and chemoprotective agents. A review, indications for use and prospects. Br J Cancer 1999;80:629-38. [PUBMED] [FULLTEXT]|
|25.||Srivastava A, Nair SC, Srivastava VM, Balamurugan AN, Jeyseelan L, Chandy M, et al. Evaluation of uroprotective efficacy of amifostine against cyclophosphamide-induced hemorrhagic cystitis. Bone Marrow Transplant 1999;23:463-7. |
|26.||Castiglione F, Mola AD, Porcile G. Protection of normal tissues from radiation and cytotoxic therapy: the development of amifostine. Tumori 1999;85:85-91. |
|27.||Wasserman T. Radioprotective effects of amifostine. Semin Oncol 1999;26:89-94. |
|28.||Vijayaraghavan R, Kulkarni A, Kumar P, Lakshmana Rao PV, Pathak U, Raza SK, et al. Prophylactic efficacy of amifostine and DRDE-07 against sulphur mustard administered through various routes, In: Pharmacological perspectives of some toxic chemicals and antidotes. Flora SJS, Romano JA, eds. New Delhi: Narosa Publishers; 2003. |