Botulinum toxin

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Botulinum toxin serotype A ( Clostridium botulinum )
Botulinum toxin serotype A (Clostridium botulinum)
Belt model according to PDB  3BTA

Existing structural data: see UniProt entry

Mass / length primary structure 447 + 848 AS (L + S)
Secondary to quaternary structure Heterodimer L + S
Cofactor Zn 2+
Precursor 1296 amino acids ; 146  kDa
Identifier
Gene name (s) BoNT / A
External IDs
Drug information
ATC code M03 AX01
DrugBank DB00083
Drug class Muscle relaxant
Transporter classification
TCDB 1.C.8
designation Botulinum / tetanus toxin family
Enzyme classification
EC, category 3.4.24.69 ,  metalloprotease
MEROPS M27.002
Response type hydrolysis
Substrate Proteins of neuroexocytosis , synaptobrevins, syntaxins
Products Fission products
Occurrence
Homology family Botulinum toxin
Parent taxon Clostridium

Botulinum toxin (BTX) , and botulinum neurotoxin ( BoNT ), Botulismustoxin , botulinum toxin , botulin - also known under the tradename Botox for the first botulinum toxin finished preparation - is a collective term for several very similar neurotoxic proteins . The neurotoxins are produced and excreted by various strains of the bacterial species Clostridium botulinum , Clostridium butyricum , Clostridium baratii and Clostridium argentinense ; they are therefore exotoxins .

The poisonous effect of these proteins is based on the inhibition of the transmission of excitation from nerve cells , which, in addition to disorders of the autonomic nervous system, can result in muscle weakness up to and including a standstill in lung function . Botulinum toxin is by far the deadliest known poison for living things like humans . The LD 50 values for mice are 3 ng / kg in the event of inhalation, up to 4 ng / kg in the case of subcutaneous ingestion and in the case of intravenous administration, even 30  p g / kg (0.03  ng / kg) are lethal.

Poisoning with botulinum toxins is called botulism and is a feared food poisoning ; it can also occur as a result of intestinal infections and wound infections with C. botulinum . Since the 1980s, the toxic proteins produced by the bacterium have been used for medical purposes, mainly for the treatment of neurological movement disorders ( dystonia ). The use in cosmetic medicine for the temporary reduction of wrinkles (duration of action 3–6 months) has been heavily criticized because of the massive increase in animal experiments caused by it. There is also a risk of serious nerve damage without a medical need for the procedure.

Clostridium botulinum occurs ubiquitously in the soil and in the sediment of water and forms resistant endospores , as does Clostridium tetani , which produces the structurally similar tetanus toxin . The evolutionary benefit of such agents pathogenic for the host to the parasitic microorganism is the subject of discussion.

history

Botulism was described scientifically for the first time in February 1815 by the Württemberg doctor and poet Justinus Kerner . In 1820 Kerner recognized the mechanism of action of the toxin as an inhibition of nerve conduction and in 1822 proposed the poison, which he called "fatty poison" and "fatty acid", in extremely low doses as a medicinal substance for various nervous disorders. The physicians Rupprecht and Müller described the poisoning as botulism for the first time in 1868 .

Clostridium botulinum (toxin type A)

The bacterium responsible for the poisoning was isolated in 1895–1897 by Emile van Ermengem , a Belgian bacteriologist, while studying ham, which was responsible for three deaths. Van Ermengem first called the microorganism Bacillus botulinus , today it is called Clostridium botulinum . Walter Kemper produced the first antiserum against botulinum toxin A as early as 1897 .

After botulinum toxins could be obtained in larger quantities from the 1920s, Carl Lammanna succeeded in 1946 in Fort Detrick ( Maryland ) at USAMRIID in the pure representation of the toxin type A. The previously suspected structure of two different protein chains was verified.

In 1949, Burgen , Dickens and Zatman in London demonstrated the inhibition of acetylcholine secretion as a cause of muscle paralysis by botulinum toxin A. A total of seven different neurotoxin serotypes (A, B, C 1 , D, E, F and G) had been isolated by around 1970 .

1973-1978, the protein was first Schantz according to one of the FDA approved method of volunteers as drug tested and Scott in 1980 by the first time to the medication of strabismus ( squint used). In 1989 the US FDA granted the first drug approval for a botulinum toxin preparation. Botulinum toxin A was approved under the name Oculinum for the indications nystagmus ("eye tremors") and blepharospasm (eyelid cramp). After the Allergan company took over the product from the then manufacturer in 1991, the trade name was changed to Botox in 1992 . In 1993, Botox was approved in Germany, as was the competitor product Dysport ( Ipsen ).

Within a few years, additional indications were spasticity of the facial muscles and legs as well as spasmodic torticollis (cervical dystonia, "torticollis").

In 1992, J. and A. Carruthers published a report on a temporary weakening effect of botulinum toxin A on skin folds (duration of action 3-6 months). Since then, the toxin has been used off-label in the cosmetic field . Preparations only received approval for this use from 2002.

Botulinum toxin B has also been used to treat cervical dystonia since 2001 .

etymology

The names for the bacterium and the toxin are derived from the Latin word botulus 'sausage', as well as the previously introduced term botulism for the clinical picture of sausage poisoning . The reason is that poisoning with botulinum toxin used to be related to sausage or canned sausages. This is the basis and the historical name sausage poison (English sausage poison ).

The bacterium Clostridium botulinum , which mainly causes botulism, can be found ubiquitously as a spore. The bacterium requires non-acidic, extremely low-oxygen environmental conditions in order to germinate and produce the toxin. Such conditions are usually given in canned sausages and vegetables as well as vacuum-packed foods; In addition, the anaerobic core zone of meat and sausage is usually an excellent substrate for the growth of Clostridium .

structure

Surface model of botulinum toxin A with light, neurotoxic chain (green) and heavy chain (blue)

Botulinum toxins are high-molecular protein complexes from the group of AB toxins , which consist of two parts: the light chain , the actual, paralyzing neurotoxin (part A) and usually the heavy chain , a non-toxic protein (part B), one Binding to nerve cells and a recording mediated. The components are connected to one another via a disulfide bridge . The lighter neurotoxic part is a zinc- containing endopeptidase . The B part binds to the presynaptic membrane of nerve cells. After uptake in synaptic vesicles , the N terminus of the heavy chain forms a membrane pore as the pH value drops. In addition, the disulfide bridge is cleaved, allowing the neurotoxic light chain to diffuse into the cytosol .

This article or the following section is not adequately provided with supporting documents ( e.g. individual evidence ). Information without sufficient evidence could be removed soon. Please help Wikipedia by researching the information and including good evidence.
pH-dependent dissociation and proteolysis protection should be demonstrated. - Ghilt ( discussion ) 18:29, 23 Jan 2020 (CET)

The B part protects the neurotoxin from proteolytic degradation in the acidic environment of the stomach after oral administration, so that the toxin remains bioavailable and can cause poisoning. However, the complex dissociates at a neutral pH value of the tissue and is no longer stable. The protein also becomes unstable after prolonged heating. In the therapeutic application of botulinum toxin, the complex proteins therefore have no essential function and do not contribute to the effectiveness. It is discussed, however, whether differences in the clinical effectiveness of the approved drugs (for example the diffusion capacity or the ability to spread) can be attributed to the different types of envelope proteins present.

Toxin types

Serologically , botulinum toxins are divided into types A to G, of which A and B are used medically. Only types A, B, E, and F are toxic to humans. The botulinum toxins are very similar in terms of their structure and biochemical effect to the tetanus toxin, which is also formed by clostridia ( Clostridium tetani ) .

Botulinum toxin serotypes
Type Some organisms in which the type is effective UniProt / UNII Variants, comments
A. Human (mostly US), chick Yes UniProt P0DPI0
UniProt P0DPI1  (A1)
UniProt Q45894  (A2)
UNII: E211KPY694

The medically used BoNT / A exists in different native, i. H. complexed forms as well as purified forms that are free of complex proteins.

B. Human (mainly Europe), "child botulism", horses, cattle Yes UniProt P10844  (B)
UniProt B1INP5  (B1)
UNII: 0Y70779M1F 		Rimabotulinum toxin B
C. Waterfowl (C1);
Cattle, horses, mink (C2)		 No UniProt P18640
UNII: FPM7829VMX 		Botulinum toxin C1 (botulinum neurotoxin C)
botulinum toxin C2
D. Cattle, poultry No UniProt P19321
UNII: 331HTW151K
E. human Yes UniProt Q00496
UniProt P30995
UNII: T579M564JY
F. human Yes UniProt A7GBG3
UniProt P30996
UNII: U1R2P71O7G
G Cases of poisoning so far unknown No UniProt Q60393
UNII: 2C3DD06QGJ 		Made by Clostridium argentinense

In addition to the seven generally recognized “classic” BoNT serotypes, several new BoNT types have been postulated. In 2013, a research group in the USA stated that they had discovered another serotype in a child's stool sample , which they called 'Type H' (BoNT / H). The structure of DNA was first kept secret despite their decryption. Investigations by the Centers for Disease Control and Prevention (CDC) later showed that it is a hybrid type with structural similarities to serotypes A and F. BoNT / X is a new serotype that is also formed by C. botulinum . A variant similar to botulin neurotoxins at the molecular level is eBoNT / J (also called BoNT / En), which is encoded by a gene from Enterococcus faecium .

Advances in DNA sequencing technology also made it possible to show differences within individual serotypes, which led to the introduction of subtypes. Such subtypes can differ by up to 36% at the amino acid level within a serotype. As of 2019, more than 40 subtypes had been described in the literature. It has been shown that the subtypes of a serotype differ in their biological activity, for example in the kinetics of uptake and substrate cleavage, the affinity to receptors or the total activity.

Mechanism of action

Botulinum toxin inhibits the transmission of excitation from nerve cells to other cells, especially at the synapses to muscle cells , which weakens or completely eliminates the contraction of the muscle . Botulinum toxins are proteins that are first produced in the bacterium as long polypeptide chains and activated by cleaved proteases . They are composed of two protein subunits , light (L, k 50 Da ) and heavy (approximately S 100 kDa) called chain.

Different functional proteins that are involved in the transmission of excitation can be destroyed by different subtypes of botulinum toxin.

The tissue specificity of the poison depends on structural features of the heavy chain . This is because botulinum toxin binds to the presynaptic part of a neuron that uses acetylcholine as a neurotransmitter - like a neuromuscular endplate . Parts of the heavy chain also convey that the poison is absorbed into the presynaptic terminal by endocytosis .

The light chain that gets into the interior of the nerve cell is the active part of the toxin. This is because it has structural features with catalytic activity and now acts as a zinc endopeptidase . This enables it to break down certain functional proteins of the cell that are needed for synaptic vesicle fusion . Through this effect, the poison hampers exocytosis as the process by which the messenger substance contained in vesicles is released into the synaptic cleft . The transmission of neuronal signals to muscle cells is thus blocked.

The membrane protein synaptobrevin (VAMP2), which is an essential component of secretory vesicles, is already broken down by the presence of a few botulinum toxin molecules of types B, D, F and G due to the catalytic effect without the botulinum toxin being consumed in the process. The botulinum toxins A and E cleave the synaptically associated protein SNAP-25 ; serotype C destroys syntaxins .

When synaptic vesicles can no longer fuse with the membrane, their transmitter acetylcholine is no longer released into the synaptic cleft. The affected nerve cell can therefore no longer sufficiently excite the assigned muscle fiber and this leads to a paralysis of the muscle on which the poison acts. When the interrupted neuromuscular transmission is reactivated, newly growing presynaptic endings temporarily take over the function of the disturbed axon terminals of the nerve cell process.

poisoning

Every year 20–40 cases of botulism are reported in Germany, of which 1–2 are fatal. "Visceral botulism" occurs rarely, which occurs as "infant botulism" or "infantile botulism" in infants and as "adult infectious botulism" in adults with a rare predisposition to be an infection. The bacterium develops from food ingested spores in the small intestine and produces the toxins there. Either a trivalent (types A, B, E) or a polyvalent antiserum (types A – G) is used as an antidote for all forms of botulism, in the USA also the heptavalent botulism antitoxin . All three are obtained from horses.

Endangered foods

First and foremost, foods that are stored under anaerobic conditions and whose environment is only slightly acidic or neutral (pH> 4.5) are at risk. The formation of the toxin can be promoted by low salt content and storage temperatures above 10 ° C, the latter often being the case with canned foods. C. botulinum also has high nutritional requirements and therefore needs a complex nutrient medium. Traditionally, canned meat and fish, mayonnaise, but also weakly acidic canned fruit or vegetables are at risk. In the case of meat products, however, (heavily) cured products are not at risk, as the nitrite contained in the curing salt inhibits the growth of C. botulinum .

Identification of contaminated products

An important indication in canning is the camber (or camber ), the curvature of the canning lid outwardly due to the generated internal pressure. Escaping gases when opening canned products also indicate contamination; in most cases they are associated with an unpleasant odor. Although such canned food must be disposed of immediately, no conclusions can be drawn from this about the infestation with botulinum toxin-producing bacteria, because the production of the poison under anaerobic conditions and without the development of gases represents a great danger. There is uncertainty as to whether the characteristics described apply , it is advisable to cook the contents of the canned food for a few minutes for safety reasons (5 minutes at 100 ° C are safe). As a heat-labile protein, the botulinum toxin is denatured and ineffective.

Symptoms in humans

The first symptoms of poisoning, known as botulism , appear after 12 to 40 hours and usually include headache and stomach ache, nausea and vomiting, as well as swallowing, speech and vision disorders, followed by muscle paralysis. In particular, paralysis of the eyes ( double vision ) and neck muscles (stiff neck) are clear indications of botulism. At this stage of the poisoning, antitoxin control may still be possible. Without treatment, death from respiratory paralysis occurs in 50% of cases after 3–6 days.

Reporting requirement

In Germany, direct or indirect evidence of the bacterium or toxin must be reported by name in accordance with Section 7 of the Infection Protection Act if the evidence indicates an acute infection.

In Switzerland, the disease botulism and the positive and negative laboratory-analytical findings of the bacterium must be reported in accordance with the Epidemics Act (EpG) in conjunction with the Epidemics Ordinance and Annex 1 and Annex 3 of the Ordinance of the EDI on the reporting of observations of communicable diseases in humans .

Preventive measures

The toxin is a protein that can be denatured , for example by heating, and thus inactivated. Its heat stability is highest at a pH of about 5.5; Even under these conditions, the botulinum toxin itself is thermally inactivated at core temperatures of over 85 ° C for 5 minutes or longer.

The spores of the toxin-producing bacteria, Clostridium botulinum , are heat-resistant and can even survive boiling water for some time. The anaerobic bacterium only grows in the absence of oxygen and not in acidic environments with a pH below 4.6. Various suitable measures can reliably prevent the bacterium from multiplying - and thus the occurrence of botulism.

sterilization

Since the sterilization technology was not very well developed in the past, it happened again and again that spores of Clostridum botulinum survived the “preservation”, ie heating of the preserves and germinated during the storage of the preserves with corresponding consequences. Today the so-called “botulinum cook” is carried out for critical products; the food is heated and kept at 121 ° C. for 3 minutes ( 3 times the F 0 value ). This also reliably kills spores.

Household (canning, canning)

  • Acidify the product sufficiently (pH <4.5)
  • Salt and brine the product
  • Store canned food at low temperatures (ideally at <5 ° C)
  • Boil or sterilize for a long enough time, possibly heating twice

Acidification in particular is an important means of inactivating C. botulinum . If the sterilization is carried out carefully and for a sufficiently long time, the spores can also be safely killed.

Industry

  • Observe general rules of industrial hygiene
  • Use as little contaminated raw material as possible
  • Implementation of the 3F concept (sterilization 3 minutes at 121 ° C, F 0 value ) or 5-minute heating at 100 ° C.

In industry, when preparing endangered foods, the bacteria and their spores are killed and the toxin is inactivated by long heating at temperatures above 100 ° C.

Therapeutic use

Indications and effectiveness

In neurology , botulinum toxin has been used as an approved drug since the early 1980s, primarily in the treatment of special movement disorders , so-called focal dystonias . These are diseases such as blepharospasm (eyelid cramp), oromandibular dystonia (mouth, tongue, pharynx), spasmodic torticollis ( torticollis ) and other cervical dystonias, graphospasm ( writer's cramp ) and spasmodic dysphonia (vocal cord spasm ). Furthermore, in segmental or secondary dystonia and spasticity, the focal symptoms can be treated with botulinum toxin. These include hemifacial spasm , movement disorders after peripheral affection of the facial nerve and certain spastic syndromes in adults and children, for example in spastic equinus , in patients with cerebral palsy , in arm spasm after a stroke or in focal spasticity of the hand and wrist after a stroke. In these indications, the intramuscular or subcutaneous injection is always used. A treatment success with improvement or temporary loss of symptoms is around 90% for eyelid cramps and between 60 and 80% for torticollis.

Further areas of application:

Certified medical training is among other things. a. offered by the Botulinum Toxin Working Group of the German Society for Neurology (DGN) and the Pain Section (IGOST) of the German Society for Orthopedics and Orthopedic Surgery ( DGOOC ).

Mechanism of action

Botulinum toxins are exotoxins produced by bacteria. If they are injected into a muscle , they block the release of the neurotransmitter acetylcholine by destroying protein complexes . As a result, the corresponding muscle can no longer be tensed as usual. Other nerve functions - such as feeling or touching - are not influenced. After a therapeutic injection , the effect slowly builds up and - depending on the indication and dose - reaches its peak after about ten days. After two to six months, the nerve endings stop sprouting and the muscles can be activated again. The injection can be performed with simultaneous measurement of an electromyogram (EMG) in the muscle, the so-called stimulation technique, ultrasound-controlled or based on anatomical knowledge. There are a number of patients with nerve-muscle diseases in whom the body forms neutralizing antibodies against subtype A after previous long and high-dose use; the effectiveness of the medication decreases or is completely lost. Subtype B preparations (Neurobloc or Myobloc, approval 2001 against torticollis syndrome ) have also been available since 2001 . However, these have a significantly shorter duration of action.

Overdose, side effects and other disadvantages

The therapeutic range of the toxin is large with an LD 50 value of about 2000 ng, determined by experiments with monkeys, for intraorbital (into the eye socket) injection; this corresponds to about 50 botox ampoules. In the event of an overdose or if the toxin gets into the bloodstream, a polyvalent botulism antitoxin from the horse is available. It is part of the emergency depot in larger hospitals . In most cases, however, the intravenous injection is given too late and the patient cannot fully recover immediately. An artificial respiration for a long time is essential.

Side effects of subcutaneous or intramuscular injection are relatively often mentioned as ptosis (during eyelid cramps), temporary muscle weakness, local pain resulting from the injection, dry mouth, disturbances in the accommodation of the eye and hematomas .

When used cosmetically, the most common reports of swallowing disorders ( dysphagia ) and, in individual cases, sarcoidosis (a disease of the connective tissue ) at the injection site and bruising due to arterial damage ( pseudoaneurysm ) were reported. In a systematic review from 2015 on known complications from the cosmetic use of botulinum toxin , it was requested that patients be informed about the following possible, serious side effects before an operation: dry eye syndrome ( keratoconjunctivitis sicca ), strabismus ( strabismus ), double vision ( diplopia ), pseudoaneurysm the superficial temporal artery ( superficial temporal artery ), neck weakness (neck weakness), voice disorder ( dysphonia ) and dysphagia .

The local injection reduces the risk of side effects (in contrast to a less useful systemic treatment), but still requires a high level of knowledge on the part of the user. So-called titration is often used, i. In other words, a dose that is too low is initially injected, and a larger amount is added every two to three weeks.

Another disadvantage is that in the best case the effect occurs after about 24 hours, normally after two to ten days, but then - depending on the dosage and indication - lasts for two to six months.

If botulinum toxin paralyzes emotional facial expressions, then also emotion-processing brain regions - such as the left amygdala - limit their activity. This prevents mimic replay and thus the understanding of emotions ("embodied emotion").

The systemic spread of botulinum toxin from the point of local application to other areas of the body, especially in the nervous system, has been proven . Although disturbances and damage have already been assigned to this spread, the underlying mechanisms of spread and its consequences have so far been virtually unexplored (as of 2016).

Dentistry

Before using botulinum toxin for the treatment of bruxism ( grinding of teeth), craniomandibular dysfunction (temporomandibular joint complaints) as well as to slim down the face, you are warned, as it can lead to massive damage to the jawbone . Botulinum toxin is injected into the masseter muscle to reduce muscle tension. A period of three months between injections, which is usually observed, is not enough to regenerate the lost bone. In some cases, the results show that the jawbone no longer regenerates even in the long term. The bone loss can lead to tooth loosening - up to and including loss of teeth - and increases the risk of fracture of the jawbone. The American Food and Drug Administration (FDA) has not yet (as of January 2017) approved the use of botulinum toxin in this area. In the current S3 guideline status 2019 on bruxism, however, the use of botulinum toxin is rated rather positively.

Injections of wrinkles in the forehead, eyes and neck area, for lip and wrinkle underfilling and for the therapy of migraine disease are permitted by licensed doctors and naturopaths , while dentists and other health care professionals and laypeople are prohibited.

In a case of violations of the ban on wrinkle injections for dentists for years, the Higher Administrative Court for the State of North Rhine-Westphalia affirmed the unreliability of a dentist, which is required for the withdrawal of his license to practice medicine. The dentist was reproached for his longstanding and persistent behavior, which he could only have eliminated by a serious change in attitude.

Approved drugs with botulinum toxin as an active ingredient

The botulinum toxin is produced as a drug by various companies:

Botulinum toxin drugs
designation Active ingredient Manufacturer Approved areas of application
Botox (D, A, CH and others) Toxin A Allergan Symptomatic treatment of various spasticities (focal spasticity of the wrist and ankle in adults, focal spasticity in patients with cerebral palsy from 2 years of age), spasms (blepharospasm, hemifacial spasm) and dystonias (cervical, focal); Relief of symptoms in chronic migraines)
Botox Cosmetics (USA)
Vistabel (D, A, CH etc.)		 Toxin A Allergan Temporary improvement in the appearance of moderate to severe wrinkles on the upper half of the face (glabellar lines, crow's feet, horizontal forehead lines) in adults with significant psychological stress
Dysport (D) Toxin A Ipsen Symptomatic treatment of various spasticity (upper extremities and ankle in adults, lower extremities in ambulatory patients with cerebral palsy from 2 years), spasms (blepharospasm, hemifacial spasm) and dystonia (cervical, focal)
Azzalure (D) Toxin A Ipsen Temporary improvement in the appearance of moderate to severe wrinkles on the upper half of the face (glabellar lines, crow's feet) in adults under 65 years of age with significant psychological stress
Xeomin (D, A, CH etc.) Toxin A Merz Wry neck (torticollis), eyelid spasm (blepharospasm) and spasticity of the upper extremities in adults
Bocouture (D, A, CH and others) Toxin A Merz Temporary improvement in the appearance of moderate to severe wrinkles on the upper half of the face (glabellar lines, crow's feet, horizontal forehead lines) in adults under 65 years of age with significant psychological stress
Nuceiva (CA, EU)
Jeuveau (USA)		 Toxin A Evolus Treatment of the wrinkles between the eyebrows (glabellar lines) in the event of psychological stress
Myobloc (USA) Toxin B Verve Treatment of cervical dystonia in adults to reduce the severity of abnormal head position and neck pain
Neurobloc (EU) Toxin B Eisai Treatment of cervical dystonia (torticollis) in adults

As of April 2009, all botulinum toxin preparations sold in the USA - as determined by the FDA based on reports of side effects - must carry a warning label. A Risk Evaluation and Mitigation Strategy (REMS) is also required for each approved product . The preparations concerned are Botox and Botox Cosmetic, Myobloc and Dysport.

The manufacturer Eisai warns against off-label use of "Neurobloc" (toxin B), as this could lead to swallowing disorders and breathing difficulties if the neurotoxin spreads to neighboring structures.

Every batch of production units of a botulinum toxin drug must be routinely tested due to its high toxicity . The classic LD 50 test is approved, in which at least 100 mice are injected with the poison into the abdomen . After 3 to 4 days, the LD 50 value is determined from the number of animals killed. The animals suffer muscle paralysis, impaired vision and shortness of breath before they suffocate. Around 600,000 mice are used around the world every year.

The following alternative test procedures are described in the European Pharmacopoeia :

  • A phrenic nerve - diaphragm - preparation in which nerves are dissected from previously killed mice or rats ( ex vivo ) and used as a test object.
  • The endopeptidase test, an in vitro molecular biological method in which the poison cleaves a synthetic protein SNAP-25.
  • A local muscle paralysis in which the poison is injected into the skin fold of the mice between the abdomen and hind leg and the extent of the paralysis of the hind leg is assessed ( determination of the effectiveness on the mouse with a paralytic endpoint ).
  • Determination in cell cultures

Further procedures are:

  • Cell culture tests with a cell line from mouse cancer cells ( neuroblastoma )
  • The immunochemical ELISA test with antibodies from test animals

The ban on animal testing for cosmetics does not apply, as botulinum toxin products are approved as drugs and are injected for use, not just applied.

Animal experiments with mice and rats have shown that botulinum toxin spreads in the central nervous system.

Extraction and storage

Botulinum toxin A is obtained from cultures of Clostridium botulinum . At a pH of 3.5, the protein is precipitated from the culture medium; the toxin is purified by a series of centrifugation , precipitation and adsorption steps. The extraction of further botulinum toxins is carried out analogously from other Clostridium species and strains. The purified toxin can be stored for long periods at −70 ° C and thawed without loss of activity. A solution for injection made from the solid protein and sterile isotonic saline solution can be stored in the refrigerator for a maximum of four hours.

Use as a warfare agent

Due to the high lethality with relatively simple production and transport, there is a risk that botulinum toxin will be used as a biological weapon . It is classified by the CDC as a substance that poses a high risk of being used in bioterrorism . Botulinum toxin could be spread in food, as an aerosol or via the drinking water supply. When dissolved in water, the toxin is colorless, odorless and tasteless.

As part of the UNSCOM inspections after the Second Gulf War , programs for the production of biological weapons were found in Iraq , among other things. a. also botulinum toxin. In Germany, botulinum toxin is subject to the War Weapons Control Act .

proof

The detection of the toxin used to be carried out in a laborious manner via a culture determination of the bacterium C. botulinum (pathogen isolation from stool or wound material); a faster detection method is RT-PCR . Direct detection of botulinum toxin from blood, stool, stomach contents or vomit is possible using bioassay methods ( ELISA or mouse bioassay), which means that the effect of samples on laboratory animals such as mice is determined.

documentary

  • Antje Christ: Botox - a poison makes a career. D, 2010, 55 minutes

literature

Web links

Wiktionary: botulinum toxin  - explanations of meanings, word origins, synonyms, translations

Remarks

Notes on the botulinum toxin serotypes table

  1. ↑ USAN : Onabotulinumtoxin A, CAS No .: 1309378-01-5
  2. ↑ USAN : Abobotulinum toxin A, CAS No .: 953397-35-8
  3. ↑ USAN : Letibotulinum toxin A, CAS no .: 1800016-51-6, UNII : W5O50S8A59
  4. ↑ USAN : Nivobotulinumtoxin A, CAS no .: 1638949-86-6
  5. ↑ USAN : Prabotulinum toxin A.
  6. ↑ USAN : Incobotulinum toxin A
  7. ↑ USAN : Daxibotulinum toxin A
  8. ↑ USAN : Evabotulinumtoxin A
  9. ↑ USAN : Rimabotulinumtoxin B, CAS No .: 93384-44-2, entry on Rimabotulinumtoxin B in the DrugBank of the University of Alberta , accessed on February 24, 2020.
  10. ↑ CAS No .: 107231-13-0
  11. ↑ CAS No .: 107231-14-1
  12. ↑ The botulinum toxin type C 2 is not a neurotoxin. Cf. Lance Simpson (ed.): Botulinum Neurotoxin and Tetanus Toxin , Academic Press, 1989, p. 7. ( limited preview in the Google book search) because there is a different mechanism of action. Botulinum toxin C 2 inhibits the formation of actin filaments , whereby the cell skeleton is damaged and actin-mediated transport processes come to a standstill.
  13. ↑ From 1959 onwards, all botulinum neurotoxin (type AG) producing clostridia were assigned to the species C. botulinum . Phenotypic and genotypic heterogeneities within the species led in 1988 to reclassification of the Clostridium botulinum type G strains to the new species C. argentinense . See JC Suen et al .: Clostridium argentinense sp.nov .: a genetically homogeneous group composed of all strains of Clostridium botulinum type G and some nontoxigenic strains previously identified as Clostridium subterminale or Clostridium hastiforme . Int. J. Sys. Bacteriol, 38, 375-381 (1988). doi: 10.1099 / 00207713-38-4-375

Individual evidence

  1. ↑ EJ Schantz, EA Johnson: Properties and use of botulinum toxin and other microbial neurotoxins in medicine. In: Microbiol Rev. 1992; 56, pp. 80-99.
  2. ↑ Kent R. Olson: Poisoning & drug overdose. 4th edition, McGraw-Hill Professional, 2004, ISBN 0-8385-8172-2 , pp. 136-138.
  3. ^ Diane O. Fleming, Debra Long Hunt: Biological Safety: Principles and Practices. ASM Press, 2000, p. 267.
  4. ↑ Entry on botulinum toxin in the ChemIDplus database of the United States National Library of Medicine (NLM) .
  5. ↑ a b c d e Botox - animal torture for a questionable beauty. In: Doctors Against Animal Experiments .
  6. ↑ K. Botrill: Growing old disgracefully: The cosmetic use of botulinum toxin. In: ATLA. 31 2003, pp. 381-391.
  7. ↑ M. Balls: Botulinum toxin testing in animals: the questions remain unanswered. In: Alternatives to laboratory animals: ATLA. Volume 31, Number 6, December 2003, pp. 611-615, PMID 15560750 (review).
  8. ^ BR Levin: The evolution and maintenance of virulence in microparasites. In: Emerging infectious diseases. Volume 2, number 2, 1996 Apr-Jun, pp. 93-102, doi: 10.3201 / eid0202.960203 , PMID 8903208 , PMC 2639826 (free full text).
  9. ↑ a b c Boris Sommer, Gerhard Sattler: Botulinum toxin in aesthetic medicine. 3rd edition, Georg Thieme Verlag, 2006, ISBN 3-13-137673-2 , pp. 1–2.
  10. ↑ Peter Moore, Markus Naumann: Handbook of botulinum toxin treatment. 2nd edition, Wiley-Blackwell, 2003, ISBN 0-632-05957-5 , p. 3.
  11. ↑ From nature: the latest discoveries in the natural sciences. Volume 31-34, 1868, p. 346.
  12. ^ A b Peter Moore, Markus Naumann: Handbook of botulinum toxin treatment. 2nd edition, Wiley-Blackwell, 2003, ISBN 0-632-05957-5 , p. 4.
  13. ↑ Martina Kerscher: Dermatocosmetics . Steinkopff Verlag, 2nd edition, 2009. ( limited preview in Google book search).
  14. ↑ Mitchell F. Brin: History of Onabotulinum Toxin A. In: Botulinum Toxin. Procedures in Cosmetic Dermatology, 3rd edition, 2013. pp. 1-7.
  15. ↑ J. Carruthers, A. Carruthers: Treatment of Glabellar Frown Lines with C. Botulinum-A Exotoxin. In: J. Dermatol. Surge Oncol. 18: (1992), pp. 17-21.
  16. ↑ JE Frampton, SE Easthope: Botulinum toxin A (Botox Cosmetic): a review of its use in the treatment of glabellar frown lines. In: American journal of clinical dermatology. Volume 4, Number 10, 2003, pp. 709-725, PMID 14507232 (review).
  17. ^ Frank J. Erbguth, Markus Naumann: Historical aspects of botulinum toxin: Justinus Kerner (1786–1862) and the "sausage poison". In: Neurology. Vol. 53, Issue 8, November 1, 1999, p. 1850.
  18. ↑ a b Entry on “botulism” in the dictionary of nutrition. Science online encyclopedias, accessed December 11, 2009.
  19. ↑ a b c d e f g h i j G. Eisenbrand (Hrsg.), P. Schreier (Hrsg.): RÖMPP Lexikon Lebensmittelchemie. 2nd edition, Thieme-Verlag, 2006, ISBN 3-13-736602-X , pp. 148-149.
  20. ↑ a b c d Entry on " botulinum toxin " in the lexicon of biology. Science online encyclopedias, accessed December 11, 2009.
  21. ^ R. Stahlmann: Botulinum toxins . Deutsche Apothekerzeitung, 34/2012 from 23 August 2012.
  22. ↑ Jason R. Barash, Stephen S. Arnon: A Novel Strain of Clostridium botulinum That Produces Type B and Type H Botulinum Toxins. J Infect Dis. (2013), October 7, 2013, doi: 10.1093 / infdis / jit449 .
  23. ↑ Botox: New toxin subject to publication ban . In: Deutsches Ärzteblatt , October 14, 2013.
  24. ^ Marieke Degen : Poison without antidote . DLF - Research News , November 6, 2013.
  25. ↑ SE Maslanka et al .: A novel botulinum toxin, previously reported as serotype H, has a hybrid structure of known serotypes A and F that is neutralized with serotype A antitoxin. J Infect Dis Volume 213 (2016) pp. 379-85 (published online in June 2015 ).
  26. ↑ UniProt P0DPK1
  27. ↑ UniProt A0A242DI27
  28. ↑ a b L. von Berg et al .: Functional detection of botulinum neurotoxin serotypes A to F by monoclonal neoepitope-specific antibodies and suspension array technology . Scientific Reports , No. 9 (2019), s. 5531 doi: 10.1038 / s41598-019-41722-z
  29. ↑ Amber Scarlatos, Bruce A. Welt, Brian Y. Cooper, Douglas Archer, Thomas DeMarse, Khe V. Chau: Methods for Detecting Botulinum Toxin with Applicability to Screening Foods Against Biological Terrorist Attacks . In: Journal of Food Science . tape 70 , no. 8 , 2005, p. 121-130 , doi : 10.1111 / j.1365-2621.2005.tb11525.x .
  30. ↑ a b Entry on “Botulism” in the Lexicon of Neurology. Science online encyclopedias, accessed December 11, 2009.
  31. ↑ J. Wang, J. Meng, M. Nugent, M. Tang, JO Dolly: Neuronal entry and high neurotoxicity of botulinum neurotoxin A require its N-terminal binding sub-domain. In: Scientific Reports . Volume 7, 03 2017, p. 44474, doi: 10.1038 / srep44474 , PMID 28295026 , PMC 5353748 (free full text).
  32. ↑ Anton de Paiva, Frédéric A. Meunier, Jordi Molgó, K. Roger Aoki, J. Oliver Dolly: Functional repair of motor endplates after botulinum neurotoxin type A poisoning: Biphasic switch of synaptic activity between nerve sprouts and their parent terminals. In: PNAS. 1999, Vol. 96, pp. 3200-3205. PMID 10077661
  33. ↑ Werner Köhler, Rainer Ansorg: Medical Microbiology. 8th edition, Elsevier, Urban & Fischer, 2001, ISBN 3-437-41640-5 , p. 402.
  34. ^ Opinion of the "DVG Specialist Group on Cattle Diseases" on the discussion on the subject of "visceral botulism" as the cause of existing diseases in cattle. (PDF) DVG (PDF; 55 kB)
  35. ↑ a b Fact sheets - Botulism. In: World Health Organization (WHO). January 10, 2018, accessed May 13, 2020 .
  36. ^ Entry on "botulinum cook". In: businessdictionary.com , accessed December 11, 2009.
  37. ↑ Michael Abele, Andres Ceballos-Baumann: Movement disorders. 2nd edition, Georg Thieme Verlag, 2005, ISBN 3-13-102392-9 , pp. 156-171.
  38. ↑ a b c Peter Berlit: Clinical Neurology. Springer, 1999, ISBN 3-540-65281-7 , pp. 902-903.
  39. ↑ a b c F. von Bruchhausen, S. Ebel, E. Hackenthal, U. Holzgrabe: Hagers manual of pharmaceutical practice. Volume 4: Substances A – K. 5th edition, Springer, 1999, ISBN 3-540-62644-1 , pp. 164-165.
  40. ^ German Society for Neurology: Migraine: bacterial poison helps the chronically ill. German Society for Neurology, press release from September 25, 2009 at Informationsdienst Wissenschaft (idw-online.de), accessed on September 15, 2015.
  41. ↑ Hartmut Göbel: The headache: causes, mechanisms, diagnostics and therapy in practice. 2nd edition, Springer, 2003, ISBN 3-540-03080-8 , pp. 481-483.
  42. ↑ Boris Sommer, Gerhard Sattler: Botulinum toxin in aesthetic medicine. 3rd edition, Georg Thieme Verlag, 2006, ISBN 3-13-137673-2 , pp. 68-73.
  43. ↑ Hamid Abdolvahab-Emminger: Exaplan, Volume 2. 5th Edition, Elsevier, Urban & Fischer, 2008, ISBN 978-3-437-42462-5 , p. 2064.
  44. ^ Gudrun Bartolome, Heidrun Schröter-Morasch: Swallowing disorders: diagnosis and rehabilitation. 3rd edition, Elsevier, Urban & Fischer, 2006, ISBN 3-437-47160-0 , pp. 215-216.
  45. ↑ Franziska Kaestner, Justine Warzok, Christian Zechmann: Crash course internal medicine: revision course on the subject catalog 3 with the incorporation of the most important examination facts. Elsevier, Urban & Fischer, 2004, ISBN 3-437-43510-8 , p. 166.
  46. ↑ a b c d S. Kahl, G. Kähler, A. Dormann: Interventional Endoskopie. Textbook and atlas. Elsevier, Urban & Fischer, 2006, ISBN 3-437-23620-2 , p. 249.
  47. ↑ New service from 2018: Botox treatment for bladder dysfunction. Retrieved February 13, 2020 .
  48. ↑ Axel Schulz, Stephan Grüner, Marcela Lippert-Grüner: Botulinum toxin - possible indications in O & U. Deutscher Ärzte-Verlag, accessed on February 13, 2020 .
  49. ↑ Use of botulinum toxin in chronic plantar fasciitis. Retrieved February 13, 2020 .
  50. ↑ Home - Igost. Retrieved February 13, 2020 .
  51. ↑ a b Boris Sommer, Gerhard Sattler: Botulinum toxin in aesthetic medicine. 3rd edition, Georg Thieme Verlag, 2006, ISBN 3-13-137673-2 , p. 15.
  52. ↑ a b E. Yiannakopoulou: Serious and long-term adverse events associated with the therapeutic and cosmetic use of botulinum toxin. In: Pharmacology. Volume 95, number 1–2, 2015, pp. 65–69, doi: 10.1159 / 000370245 , PMID 25613637 (free full text) (review).
  53. ↑ EP Sorensen, C. Urman: Cosmetic complications: rare and serious events following botulinum toxin and soft tissue filler administration. In: Journal of drugs in dermatology: JDD. Volume 14, Number 5, May 2015, pp. 486-491, PMID 25942667 (review).
  54. ↑ Andreas Henn Lotter, Christian Dresel, Florian Castrop, Andres O. Ceballos-Baumann, Afra M. Wohlschläger, Bernhard Haslinger: The link between facial feedback and Neural Activity within Central Circuit Ries of Emotion-New Insights from botulinum toxin induced denervation of Frown Muscles . Cereb. Cortex, Volume 19, Issue 3, pp. 537-542; doi: 10.1093 / cercor / bhn104 .
  55. ↑ KG Raphael, A. Tadinada u. a .: Osteopenic consequences of botulinum toxin injections in the masticatory muscles: a pilot study. In: Journal of Oral Rehabilitation. 41, 2014, p. 555, doi: 10.1111 / joor.12180 .
  56. ↑ Is Botox Safe and Effective for TMD? The TMJ Association, Retrieved June 13, 2016.
  57. ↑ AWMF: S3 guideline for the diagnosis and treatment of bruxism. (PDF) AWMF, accessed on February 13, 2020 .
  58. ↑ Decision of May 17, 2017 - Ref .: 13 A 168/16. Higher Administrative Court of North Rhine-Westphalia
  59. ↑ FDA Requires Boxed Warning for All Botulinum Toxin Products. FDA News & Events, April 30, 2009.
  60. ↑ Notice to members of the health professions on the risks associated with the "off-label" use of NeuroBloc (botulinum toxin type B). (PDF) In: akdae.de , February 25, 2013.
  61. ↑ S. Bitz: The Botulinum Neurotoxin LD 50 Test. Problems and Solutions. ( Memento of April 7, 2014 in the Internet Archive ) (PDF; 212 kB) Altex 27, 2/2010, pp. 114–116.
  62. ↑ Botox mice: agonizing death for fewer wrinkles. In: Kassensturz , broadcast on November 20, 2007.
  63. ↑ European Pharmacopoeia, 8th edition, Grundwerk 2014, pp. 2538 f., Monograph botulinum toxin type A for injection
  64. ↑ Study Finds Botulinum Toxin Spreads to CNS Tissue in Mice. In: journals.lww.com
  65. ↑ Gisela Telis: Have Botox, Will Travel. ( Memento of September 13, 2011 in the Internet Archive ) In: sciencemag.org , April 2, 2008.
  66. ↑ Botulinum Toxin (Botulism). Accessed March 7, 2018 (English).
  67. ^ Biological and Chemical Terrorism: Strategic Plan for Preparedness and Response. Retrieved March 7, 2018 .
  68. ↑ Bernd D. Domres, Andreas Manger, Rainer Wenke, Stefan Brockmann, Mike Kay, Horst Miska: GEMAESS - CBRN Hazardous Situations . Ed .: Johanniter-Unfall-Hilfe eV 2010, p. 89 ( Online (PDF)).
  69. ↑ John Pike: Botulinum Toxins - Biological Weapons. Accessed March 7, 2018 (English).
  70. ↑ H. Russmann: Toxins . In: Federal Health Gazette - Health Research - Health Protection . tape 46 , no. 11 , November 1, 2003, p. 989-996 , doi : 10.1007 / s00103-003-0716-0 ( online [PDF]). Online ( Memento from March 8, 2018 in the Internet Archive )
  71. ↑ War Weapons Control Act, Annex (to Section 1, Paragraph 1) , II. Biological Weapons, 3.1 c) Clostridium botulinum 3.1 d) Clostridium botulinum toxins.
  72. ↑ U. Messelhäusser, R. Zucker, H. Ziegler, D. Elmer-Englhard, W. Kleih, C. Höller, U. Busch: Detection of Clostridium botulinum types A, B, E and F using real-time PCR. (Abstract) . In: Journal for Consumer Protection and Food Safety , May 2007, Vol. 2, Iss. 2, pp. 198-201; doi: 10.1007 / s00003-007-0174-y .
  73. ↑ Clostridium botulinum (botulism) . Health Center North Rhine-Westphalia; Retrieved March 5, 2013.
  74. ↑ Botox - A poison makes a career. Documentation Germany 2010.