Molecular medicines for neutralization of Clostridium botulinum neurotoxin

Paper Details

Review Paper 01/06/2019
Views (286) Download (17)

Molecular medicines for neutralization of Clostridium botulinum neurotoxin

Kausar Malik, Mujahid Hussain, Rida Sadaqat, Hassan Ahmad, Muhammad Hamza Basit Shafiq Azam, Hasnain Qamar, Arshia Nazir, Haleema Sadia, Asma Arshad
Int. J. Biosci.14( 6), 78-90, June 2019.
Certificate: IJB 2019 [Generate Certificate]


Botulism is characterized by symmetrical, descending, flaccid paralysis of motor and autonomic nerves, caused by the spore-forming, obligate anaerobic bacterium Clostridium botulinum. Strains of Clostridium botulinum are known to produce the most poisonous neurotoxins in mankind. Clostridium botulinum produces seven genetically distinct neurotoxins known as BoNT/A, BoNT/B, BoNT/C, BoNT/D, BoNT/E, BoNT/F and BoNT/G.All the serotypes share same structure and molecular weight, but differ in their cellular substrate and target cleavage site. Botulinum toxins work by blocking the release of acetylcholine in four stages, binding, internalization, translocation and inhibition. The food borne botulism, wound botulism, infant botulism and adult botulism are main clinical features. Different molecular techniques like Mouse lethality assay, ELISA, immuno-PCR, chemiluminescent   slot   blot immunoassay, electrochemiluminenscence, radioimmunoassay, lateral flow immunoassays and End peptidase assay are mostly used to detect the BoNTs. Antitoxins such as BabyBIG, Equine, Mabs and HBAT are used for treatment of BoNTs intravenously or intra- muscularly. At molecular level Peptide Based Inhibitor, Phage display technology and Aptamers are used. A proper delivery system is required to deliver inhibitors to target nerves to reverse the clinical effects. Heavy chain of BoNT has been shown to be the natural, safe and potential delivery system to deliver inhibitory molecules in the affected nerves.


Martin L, Cornille F, Turcaud S, Roques BP. 1999. Metallopeptidase Inhibitors of Tetanus Toxin: A Combinatorial Approach. Journal of Medical Chemistry 42, 515–525.

Aoki KR, Guyer B. 2001. Botulinum toxin type A and other botulinum toxin serotypes: a comparative review of biochemical and pharmacological actions. European Journal of Neurology 8, 21–29.

Barash JR, Arnon SS. 2014. A Novel Strain of Clostridium botulinum That Produces Type B and Type H Botulinum Toxins. Journal of Infectious Diseases 209, 183–191. 10.1093/infdis/jit449

Black DJ, Dolly JO. 1987. Selective location of acceptors for botulinum neurotoxin a in the central and peripheral nervous systems. Neuroscience 23, 767-779.

Blasil J, Chapman ER, Yamasaki S, Binz T, Niemann H, Jahn R. 1993. Botulinum neurotoxin C1 blocks neurotransmitter release by means of cleaving HPC-1 / syntaxin. The EMBO Journal 12, 4821–4828.

Blaustein R, Germann WJ, Finkelstein A, Dasgupta BR. 1987. The N-terminal half of the heavy chain of botulinurn type A neurotoxin forms channels in planar phospholipid bilayers. Elsevier Science Publishers 226, 115–120.

Cai S, Singh BR. 2014. Strategies to Design Inhibitors of Clostridium Botulinum Neurotoxins Strategies to Design Inhibitors of Clostridium Botulinum Neurotoxins. Infectious Disorders – Drug Targets 7, 47-57.

Dolly JO, Aoki KR. 2006. The structure and mode of action of different botulinum toxins. European Journal of Neurology 13, 1–9.

Dolly O. 2003. Synaptic Transmission : Inhibition of Neurotransmitter Release by Botulinum Toxins. Headache 43, 16–24.

Goodnough MC, Oyler G, Fishman PS, Johnson EA, Neale EA, Keller JE, Tepp WH, Clark M, Hartz S, Ã MAY. 2002. Development of a delivery vehicle for intracellular transport of botulinum neurotoxin antagonists. FEBS Letters 513, 163–168

Grenda T, Kukier ELŻB, Sieradzki Z, Goldsztejn M, Kwiatek K. 2012. IN-House validation of multiplex pcr method for detection of clostridium botulinum in food and feed. Bulletin of the Veterinary Institute in Pulawy 56, 155–160.

Hayden J, Pires J, Hamilton M, Moore G. 2000. Novel inhibitors of botulinus neurotoxin “A” based on variations of the SNARE motif. Proceedings of Western Pharmacology Society 43, 71-74.

Hayden J, Pires J, Roy S, Hamilton M, Moore GJ. 2003. Discovery and Design of Novel Inhibitors of Botulinus Neurotoxin A : Targeted ‘ Hinge ’ Peptide Libraries. Journal of Applied Toxicology 23, 1–7.

Kongsaengdao S, Samintarapanya K, Rusmeechan S, Wongsa A, Pothirat C, Permpikul C, Pongpakdee S, Puavilai W, Kateruttanakul P, Phengtham U, Panjapornpon K, Janma J, Piyavechviratana K, Sithinamsuwan P, Deesomchok A, Tongyoo S, Vilaichone W, Boonyapisit K, Mayotarn S, Piya-isragul B, Rattanaphon A, Intalapaporn P, Dusitanond P, Harnsomburana P, Laowittawas W, Chairangsaris P, Suwantamee J, Wongmek W, Ratanarat R, Poompichate A, Panyadilok H, Sutcharitchan N, Chuesuwan A, Oranrigsupau P, Sutthapas C, Tanprawate S, Lorsuwansiri J, Phattana N, Botulism T, Group S, Hospital N. 2006. An Outbreak of Botulism in Thailand : Clinical Manifestations and Management of Severe Respiratory Failure. Clinical Infectious Diseases 43, 1247–1256.

Li L, Ram B. 1999. Structure-Function Relationship of Clostridial Neurotoxins. Journal of Toxicology-Toxin Reviews 18, 95–112.

Montecucco C. 1986. How do tetanus and botulinum toxins bind to neuronal membranes? Trends in biochemical sciences 11, 314-317.

Montecucco C, Papini E, Schiavo G. 1994. Bacterial protein toxins penetrate cells via a four-step mechanism. FEBS Letters 346, 92–98.

Montecucco C, Rossetto O, Schiavo G. 2004. Presynaptic Receptor Arrays for Clostridial Neurotoxins. Trends in Microbiology 12, 442-446.

Nimjee SM, Rusconi CP, Sullenger BA. 2005. Aptamers: an emerging class of therapeutics. Annual Review of Medicine 56, 555-583.

Peck M. 2006. Clostridium botulinum and the safety of minimally heated, chilled foods: an emerging issue. Journal of Applied Microbiology 101, 556-570.

Pestourie C, Tavitian B, Duconge F. 2005. Aptamers against extracellular targets for in vivo applications. Biochimie 87, 921-930.

Rossetto O, Schiavo G, Monteccucco C, Poulain B, Deloye F, Lozzi L, Shone CC. 1994. SNARE motif and neurotoxins. Nature 372, 415-416.

Schmidt JJ, Stafford RG. 2002. A high‐affinity competitive inhibitor of type A botulinum neurotoxin protease activity. FEBS Letters 532, 423-426.

Schmidt JJ, Stafford RG, Millard CB. 2001. High-throughput assays for botulinum neurotoxin proteolytic activity: serotypes A, B, D, and F. Analytical Biochemistry 296, 130-137.

Tomic M, Garcia C, Lou J, Geren IN, Meng Q, Conrad F, Wen W, Smith TJ, Brown J, Smith LA, Wajid A. 2013. Recombinant monoclonal-antibody- based antitoxins for treatment of types A, B, and E botulism. Toxicon 68, 99-100.

Tuerk C, Gold L. 1990. Systematic evolution of ligands by exponential enrichment: RNA ligands to bacteriophage T4 DNA polymerase. Science 249, 505-510.

United States. Centers for Disease Control and Prevention.” Botulism.” Apr. 25, 2014.

Zdanovskaia MV, Los G, Zdanovsky AG. 2000. Recombinant Derivatives of Clostridial Neurotoxins as Delivery Vehicles for Proteins and Small Organic Molecules. Journal of Protein Chemistry 19, 699-707.

Zdanovsky AG, Karassina NV, Simpson D, Zdanovskaia MV. 2001. Peptide phage display library as source for inhibitors of clostridial neurotoxins. Journal of Protein Chemistry 20, 73-80.

Arnon SS. 1998. Infant botulism. In: Feigen RD, Cherry JD, Eds. Textbook of Pediatric Infectious Diseases, 4th ed, Philadelphia: WB Saunders, 1570-1577.

Centers for Disease Control and Prevention. Botulism in the United States. 1899-1973.  Handbook for Epidemiologists, Clinicians, and Laboratory Workers. Atlanta: Centers for Disease Control; 1978, 3. Publication no. (CDC) 74-8279/G.

Chia JK, Clark JB, Ryan CA, Pollack M. 1983. Botulism in an adult associated with foodborne intestinal infection with Clostridium botulinum. The New England Journal of Medicine 315, 239-241.

Griffin PM, Hatheway CL, Rosenbaum RB, Sokolow R. 1997. Endogenous antibody production to botulism toxin in an adult with intestinal colonization botulism and underlying Crohn’s disease. Journal of Infectious Diseases 175, 633-637.

Hughes JM, Blumenthal JR, Merson MH, Lombard GL, Dowell VRJ, Gangarosa EJ. 1981. Clinical features of types A and B food-borne botulism. Annals of Internal Medicine 95, 442- 445.

McCroskey LM, Hatheway CL. 1988. Laboratory findings in four cases of adult botulism suggest colonization of the intestinal tract. Journal of Clinical Microbiology 26, 1052-1054.

Merson MH, Dowell VRJ. 1973. Epidemiologic, clinical and laboratory aspects of wound botulism. The New England Journal of Medicine 289, 1105-1110. 

Spika JS, Shaffer N, Hargrett-Bean N, Collin DS, MacDonald KL, Blake PA. 1979. Risk factors for infant botulism in the United States. The American Journal of Diseases of Children 143, 828-832.

United States. 1978. MMWR Morbidity and Mortality Weekly Report 28, 73-75.

Louis ME, Peck SHS, Bowering D. 1988. Botulism from chopped garlic: delayed recognition of a major outbreak. Annals of Internal Medicine 108, 363-368.

McNally RE, Morrison MB, Berndt JE, Fisher JE, Bo-Berry JI, Packett VE. 1994. Effectiveness of Medical Defense Interventions against Predicted Battlefield Levels of Botulinum Toxin A. CorpJoppa MD: Science Applications International.

Hatheway CL. 1995. Botulism: the present status of the Disease. Clostridial Neurotoxins 195, 55-75.

Wilson R, Morris JGJ, Snyder JD, Feldman RA. 1982. Clinical characteristics of infant botulism in the United States: a study of the non-California cases. Pediatric Infectious Diseases 1, 148-150.

Woodruff BA, Griffin PM, McCroskey LM, Smart JF, Wainwright RB, Bryant RG. 1992. Clinical and laboratory comparison of botulism from toxin types A, B and E in the United States. Journal of Infectious Diseases 166, 1281-1286.