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1.
J Neurol ; 270(3): 1524-1530, 2023 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-36434127

RESUMO

Botulinum toxin (BT) therapy may be blocked by antibodies (BT-AB) resulting in BT-AB induced therapy failure (ABF). BT-AB may be detected by the mouse lethality assay (MLA), the mouse diaphragm assay (MDA) and the sternocleidomastoid test (SCMT). For the first time, we wanted to compare all three BT-AB tests and correlate them to subjective complaint of complete or partial secondary therapy failure in 37 patients with cervical dystonia (25 females, 12 males, age 51.2 ± 11.4 years, disease duration 12.4 ± 6.3 years). Complaint of therapy failure was not correlated with any of the BT-AB tests. MDA and MLA are closely correlated, indicating that the MDA might replace the MLA as the current gold standard for BT-AB measurement. The SCMT is closely correlated with MDA and MLA confirming that BT-AB titres and BT's paretic effect are in a functional balance: low BT-AB titres are reducing BT's paretic effect only marginally, whereas high BT-AB titres may completely block it. When therapy failure is classified as secondary and permanent, BT-AB evaluation is recommended and any BT-AB test may be applied. For MDA > 10 mU/ml, MLA > 3 and SCMT < 25%, ABF is highly likely. MDA < 0.6 mU/ml are therapeutically irrelevant. They are neither correlated with pathologic MLA nor with pathologic SCMT. They should not be the basis for treatment decisions, such as switching dystonia therapy to deep brain stimulation. All other results are intermediate results. Their interactions with therapy efficacy is unpredictable. In these cases, BT-AB tests should be repeated or one or two additional test methods should be applied.


Assuntos
Toxinas Botulínicas Tipo A , Distúrbios Distônicos , Torcicolo , Masculino , Feminino , Animais , Camundongos , Falha de Tratamento , Anticorpos , Torcicolo/tratamento farmacológico
2.
J Neurol ; 270(2): 788-796, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36195775

RESUMO

BACKGROUND: For many indications, BoNT/A is repetitively injected with the risk of developing neutralizing antibodies (NABs). Therefore, it is important to analyze whether there is a difference in antigenicity between the different licensed BoNT/A preparations. METHODS: In this cross-sectional study, the prevalence of NABs was tested by means of the sensitive mouse hemidiaphragm assay (MHDA) in 645 patients. Patients were split into those having exclusively been treated with the complex protein-free incoBoNT/A preparation (CF-MON group) and those having started BoNT/A therapy with a complex protein-containing BoNT/A preparation (CC-I group). This CC-I group was split into those patients who remained either on abo- or onaBoNT/A (CC-MON group) and those who had been treated with at least two BoNT/A preparations (CC-SWI group). To balance treatment duration, only CC-MON patients who did not start their BoNT/A therapy more than 10 years before recruitment (CC-MON-10 group) were further analyzed. The log-rank test was used to compare the prevalence of NABs in the CF-MON and CC-MON-10 group. RESULTS: In the CF-MON subgroup, no patient developed NABs. In the CC-I group, 84 patients were NAB-positive. NABs were found in 33.3% of those who switched preparations (CC-SWI) and in 5.9% of the CC-MON-10 group. Kaplan-Meier curves for remaining NAB-negative under continuous BoNT/A therapy were significantly different (p < 0.035) between the CF-MON and CC-MON-10 group. CONCLUSION: Frequent injections of a complex protein-containing BoNT/A preparation are associated with significantly higher risks of developing NABs than injections with the same frequency using the complex protein-free incoBoNT/A preparation.


Assuntos
Toxinas Botulínicas Tipo A , Fármacos Neuromusculares , Animais , Camundongos , Estudos Transversais , Anticorpos Neutralizantes
3.
Toxicology ; 481: 153341, 2022 11.
Artigo em Inglês | MEDLINE | ID: mdl-36191878

RESUMO

Like all proteins foreign to the human body, also botulinum toxin (BT) is antigenic and may stimulate an immune response with formation of antibodies (BT-AB). Affected patients may no longer respond to BT therapy and various degrees of BT-AB related therapy failure (ABF) may result. We want to review the immunological interactions between BT and BT-AB, the prevalence, the time course and the risk factors for BT-AB formation as they are related to the treatment algorithms, the patient's immune system and to exogenic factors. Special emphasis is placed on various features of the BT drugs including the specific biological activity (SBA) as a predictor of their antigenicity. Quantitative detection of BT-AB by the mouse diaphragm assay will be demonstrated. As ABF may have serious consequences for patients affected, careful risk factor analysis is warranted to reduce them wherever possible.


Assuntos
Toxinas Botulínicas Tipo A , Camundongos , Animais , Humanos , Toxinas Botulínicas Tipo A/uso terapêutico , Toxinas Botulínicas Tipo A/toxicidade , Falha de Tratamento , Bioensaio , Diafragma , Fatores de Risco
4.
Ann Clin Transl Neurol ; 8(1): 15-28, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-33259153

RESUMO

OBJECTIVE: The objective of the study was the analysis of adherence and self-perceived treatment response to long-term botulinum neurotoxin type A (BoNT-A) treatment in different neurological indications. METHODS: In this retrospective, monocentric, observational study, cross-sectional and longitudinal data of 1351 patients documenting 20705 injection appointments at the BoNT outpatient clinic of Heinrich Heine University Duesseldorf between 1989 and 2014 were retrospectively analyzed. Patients had been treated with BoNT for neurological conditions, including cervical dystonia (CD), blepharospasm (BSP), other dystonia (ODT), hemifacial spasm (HFS), and spasticity (SPAS). The parameters longitudinally analyzed for the entire cohort were therapy duration as well as the mean and cumulative BoNT-A dose. Cross-sectionally, for subgroups of at least 721, patients' global self-perceived quality of health and life, global self-perceived reduction of symptoms by BoNT-A treatment as well as the clinical global impression were evaluated. Furthermore, mouse hemidiaphragm assay antibodies (MHDA-ABs) were analyzed in a subgroup. RESULTS: The mean treatment duration was 4.58 years (95% CI 4.32-4.84), and 678 (50.2%) therapy dropouts of 1351 patients occurred within the first 8 years. Therapy adherence and self-perceived symptom reduction in long-term BoNT-A treatment over the years were significantly longer in BSP, HFS, and CD patients than in ODT and SPAS patients. INTERPRETATION: The treatment indication determines long-term adherence and self-perceived symptom reduction in BoNT-A therapy, which are better in BSP, HFS, and CD patients than in ODT and SPAS patients. MHDA-ABs had a significant impact on global self-perceived symptom reduction, but with only a limited degree.


Assuntos
Toxinas Botulínicas Tipo A/uso terapêutico , Adesão à Medicação/estatística & dados numéricos , Fármacos Neuromusculares/uso terapêutico , Adulto , Idoso , Blefarospasmo/tratamento farmacológico , Distonia/tratamento farmacológico , Feminino , Espasmo Hemifacial/tratamento farmacológico , Humanos , Masculino , Pessoa de Meia-Idade , Espasticidade Muscular/tratamento farmacológico , Estudos Retrospectivos
5.
J Neural Transm (Vienna) ; 126(12): 1625-1629, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31707463

RESUMO

LanbotulinumtoxinA (LAN) is manufactured and registered in China since 1994. Despite its widespread use in China and its increasing use in other Asian countries and in South America, it is not yet well known elsewhere. We wanted to compare its potency labelling using the mouse diaphragm assay (MDA), an isolated muscle model for botulinum toxin (BT) potency measurements, which is superior to clinical tests and which was recently refined as an alternative batch release assay for BT manufacturing. We also wanted to estimate LAN manufacturing quality by testing its inter-batch potency consistency. Potencies of 20, 60 and 100 MU of LAN, onabotulinumtoxinA (ONA) and incobotulinumtoxinA (INCO) were measured by the inversely related paresis time (PT) in the MDA. The PT (M ± SD) of all doses of LAN, ONA and INCO was 90.4 ± 27.0 min, 114.9 ± 46.5 min and 94.3 ± 29.9 min, respectively. Statistical analysis demonstrated indistinguishable potency labelling of LAN and INCO, but revealed a slightly lower potency of ONA compared to LAN and INCO. PT of LAN batch 1 and LAN batch 2 was 86.9 ± 21.2 min and 94.0 ± 32.8 min, respectively (no statistically significant difference), suggesting an adequate LAN manufacturing consistency. The MDA is an appropriate instrument for potency testing of BT drugs, including new ones currently under development. Our results allow comparing therapeutic effects, adverse effects and economics of LAN, ONA and INCO. They also suggest adequate manufacturing consistency of LAN.


Assuntos
Toxinas Botulínicas Tipo A/farmacologia , Diafragma/efeitos dos fármacos , Fármacos Neuromusculares/farmacologia , Animais , Camundongos , Técnicas de Cultura de Órgãos
8.
Toxicon ; 168: 158-163, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31323228

RESUMO

Botulinum neurotoxin (BoNT) is synthesized as a progenitor toxin complex (PTC) by Clostridium botulinum. This PTC comprises, in addition to the neurotoxin itself, neurotoxin associated proteins (NAPs) which are composed of three hemagglutinins and one non-toxic, non-hemagglutinin protein. After oral ingestion, these NAPs protect the neurotoxin from the low pH and proteases in the gastrointestinal tract and play a role in the entry via the intestinal barrier. Two of the three therapeutically used botulinum neurotoxin serotype A (BoNT/A) products (onabotulinumtoxinA and abobotulinumtoxinA) contain different amounts of NAPs, while incobotulinumtoxinA, lacks these proteins. In addition, human serum albumin (HSA) that is supposed to stabilize BoNT/A is added at different concentrations. Up to now, the function of the NAPs and HSA after parenteral therapeutic application is not completely understood. To investigate the influence of NAPs and HSA on potency of BoNT/A, we used the ex vivo mouse phrenic nerve hemidiaphragm assay. Increasing doses of HSA resulted dose-dependently in a more pronounced effect of BoNT/A. Though, a plateau was reached with concentrations of 0.8 mg/ml HSA and higher, the accessory addition of NAPs in a relevant amount (4 ng/ml) did not further enhance the effect of BoNT/A. In conclusion, in our ex vivo assay an adequate concentration of HSA prevented BoNT/A from loss of effect and supplementary NAPs did not alter this effect. A confirmation of these data in an in vivo assay is still lacking. However, it might be supposed that even in clinically applied BoNT/A products an increase of HSA accompanied by the avoidance of NAPs could potentially reduce the injected dose and, thus, the risk of unwanted side effects, the treatment costs as well as the risk of a secondary therapy failure due to BoNT/A neutralizing antibodies.


Assuntos
Toxinas Botulínicas Tipo A/farmacologia , Albumina Sérica Humana/farmacologia , Animais , Toxinas Botulínicas Tipo A/química , Masculino , Camundongos , Complexos Multiproteicos/química , Complexos Multiproteicos/farmacologia , Nervo Frênico/efeitos dos fármacos
9.
J Neural Transm (Vienna) ; 126(8): 1047-1050, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31168666

RESUMO

Botulinum toxin (BT) consists of botulinum neurotoxin and complexing proteins (CPs). CPs might provide mechanical protection for botulinum neurotoxin. As incobotulinumtoxinA (INCO, Xeomin®) does not contain CPs, we wanted to compare its mechanical stability to that of onabotulinumtoxinA (ONA, Botox®) containing CPs. For this, ONA and INCO were reconstituted without mechanical stress (NS) and with mechanical stress (WS) generated by a recently introduced stress test. Potencies were then measured by the paralysis times (PTs) in the mouse diaphragm assay. ONA-PT was 75.8 ± 10.3 min (n = 6) under NS and 116.7 ± 29.8 min (n = 6) under WS (two-tailed t test, p = 0.002). Mechanical stress increased the ONA-PT by 35.0% on the Growth Percentage Index. INCO-PT was 66.0 ± 7.0 min for NS and 76.0 ± 1.0 min for WS (t test, p = 0.129). Mechanical stress increased the INCO-PT by 13.2% on the Growth Percentage Index. Our data show that mechanical stress inactivates a CP-containing BT drug, but not a CP-free BT drug. We conclude that CPs do not provide protection against mechanical stress, supporting the view that CPs are not necessary for therapeutic purposes.


Assuntos
Toxinas Botulínicas Tipo A/química , Animais , Toxinas Botulínicas Tipo A/farmacologia , Diafragma/efeitos dos fármacos , Camundongos , Movimento/efeitos dos fármacos , Neurotoxinas/farmacologia , Estabilidade Proteica , Estresse Mecânico
10.
J Neural Transm (Vienna) ; 126(10): 1337-1340, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31203435

RESUMO

Based on epidemiological data it was believed that botulinumtoxin type D (BT-D) may not block human cholinergic synapses. We wanted to investigate BT-D's effect on the autonomic cholinergic synapse in humans. For this, we compared in four volunteers intraindividually the hypohidrotic effect of intradermal BT-D and BT-A in Minor's iodine starch sweat test. Altogether, we studied BT-D in doses of 4, 8, 16 and 32MU and BT-A in doses of 2, 4, 8 and 16MU at weekly intervals throughout a period of 13 weeks. All BT doses were diluted in 0.2 ml 0.9% NaCl/H2O. Overall 704 data points were collected. Combined over all four subjects and all four doses BT-D's hypohidrotic effect intensity was half of BT-A's. BT-D's effect peaked around 5 weeks, BT-A's around 7 weeks. BT-D's effect duration was around 12 weeks, of BT-A's was around 14 weeks. For both BT types the hypohidrotic effect was dose dependent. BT-D, when injected intradermally, can block autonomic cholinergic synapses in humans. Compared to BT-A, BT-D's effect intensity was half and its effect duration was some 2 weeks shorter. With its weaker and shorter effect BT-D does not seem to promise therapeutic effects superior to BT-A.


Assuntos
Inibidores da Liberação da Acetilcolina/administração & dosagem , Toxinas Botulínicas/administração & dosagem , Neurônios Colinérgicos/efeitos dos fármacos , Hipo-Hidrose/induzido quimicamente , Sinapses/efeitos dos fármacos , Inibidores da Liberação da Acetilcolina/toxicidade , Adulto , Toxinas Botulínicas/toxicidade , Neurônios Colinérgicos/fisiologia , Relação Dose-Resposta a Droga , Humanos , Hipo-Hidrose/diagnóstico , Masculino , Pessoa de Meia-Idade , Sinapses/fisiologia
11.
Clin Neurophysiol ; 130(6): 1066-1073, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30871800

RESUMO

OBJECTIVES: Botulinum neurotoxin serotypes A and B (BoNT/A & B) are highly effective medicines to treat hyperactive cholinergic neurons. Due to neutralizing antibody formation, some patients may become non-responders. In these cases, the serotypes BoNT/C-G might become treatment alternatives. BoNT/D is genetically least related to BoNT/A & B and thereby circumventing neutralisation in A/B non-responders. We produced BoNT/D and compared its pharmacology with BoNT/A ex vivo in mice tissue and in vivo in human volunteers. METHODS: BoNT/D was expressed recombinantly in E. coli, isolated by chromatography and its ex vivo potency was determined at mouse phrenic nerve hemidiaphragm preparations. Different doses of BoNT/D or incobotulinumtoxinA were injected into the extensor digitorum brevis (EDB) muscles (n = 30) of human volunteers. Their compound muscle action potentials were measured 11 times by electroneurography within 220 days. RESULTS: Despite a 3.7-fold lower ex vivo potency in mice, a 110-fold higher dosage of BoNT/D achieved the same clinical effect as incobotulinumtoxinA while showing a 50% shortened duration of action. CONCLUSIONS: BoNT/D blocks dose-dependently acetylcholine release in human motoneurons upon intramuscular administration, but its potency and duration of action is inferior to approved BoNT/A based drugs. SIGNIFICANCE: BoNT/D constitutes a potential treatment alternative for BoNT/A & B non-responders.


Assuntos
Toxinas Botulínicas Tipo A/administração & dosagem , Toxinas Botulínicas/administração & dosagem , Músculo Esquelético/efeitos dos fármacos , Fármacos Neuromusculares/administração & dosagem , Adulto , Animais , Humanos , Masculino , Camundongos , Músculo Esquelético/fisiologia , Resultado do Tratamento
12.
Ther Adv Neurol Disord ; 12: 1756286419892078, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31897089

RESUMO

BACKGROUND: The aim of the study was to test the clinical relevance of neutralizing antibodies (NABs) in patients with cervical dystonia (CD) still responding to repeat injections with botulinum toxin type A (BoNT/A). METHODS: Enzyme-linked immunosorbent assay (ELISA)-test evidence from a cross-sectional study on 221 CD-patients with treatment durations of between 2 and 21 years and still responding to repeat BoNT/A-injections showed the presence of antibodies against BoNT/A in 39 patients. A mouse hemi-diaphragm (MHDA) confirmation test was performed in these 39 ELISA-positive patients, and demographic (age, sex, age at onset of CD) and treatment-related (duration of treatment, mean dose of the last 10 injections, TSUI-score, patient's subjective scoring of the treatment effect, patient's scoring of quality of life by means of the CDQ24-questionnaire) data from these 39 patients were compared with data from ELISA-negative patients. Paralysis time, the MHDA outcome measure, was correlated with clinical data. RESULTS: The ELISA-positive CD-patients had significantly higher TSUI-scores (p < 0.015), and had been treated for significant longer (p < 0.022) and with significantly higher doses (p < 0.001). Patient's rating of BoNT/A-treatment effect and quality of life tended to be worse in ELISA-positive compared with ELISA-negative patients. The paralysis time of ELISA-positive patients was significantly correlated with the mean dose of the last 10 injections (p < 0.027) and the pain subscore of the CDQ24 (p < 0.012). CONCLUSIONS: Presence of NABs is clinically relevant in CD, leading to a significantly worse head position, therapy with significantly higher BoNT/A doses, and a correlation between the CDQ24 pain-subscore and antibody titers.

13.
Neurology ; 92(1): e48-e54, 2019 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-30464031

RESUMO

OBJECTIVE: To investigate the prevalence of neutralizing antibodies (NAbs) against botulinum neurotoxin type A (BoNT/A) during long-term BoNT/A treatment in different neurologic indications. METHODS: In this monocentric, observational cross-sectional study, 596 outpatients treated with BoNT/A for different indications were tested for BoNT/A binding antibodies by ELISA. Positive samples were investigated for NAbs with the mouse hemidiaphragm test. The prevalence of NAbs was analyzed for different indications: facial hemispasm, blepharospasm, cervical dystonia, other dystonia, and spasticity. Besides the rate of NAb-positive patients overall and per patient subgroup, a Kaplan-Meier analysis of the probability of remaining NAb negative with duration of treatment is provided, and a stepwise binary logistic regression analysis is performed to identify factors significantly contributing to the induction of NAbs. RESULTS: Overall, 83 of 596 patients (13.9%) had measurable NAbs. The probability of developing NAbs increased with the single and cumulative dose of treatment and was influenced by the BoNT/A formulation, while all other factors analyzed, including disease entity and treatment duration, had no additional influence. CONCLUSIONS: We present the largest study to date of the prevalence of BoNT/A NAbs in a large unbiased cohort of patients including the relevant neurologic indications. Repeated injections of BoNT/A inevitably bear the risk of developing NAbs. However, in addition to avoiding booster injections and providing short intervals between injections, reducing the individual injected doses may diminish the risk of NAb induction independently of the indication for which BoNT/A is used.


Assuntos
Anticorpos Neutralizantes/sangue , Blefarospasmo/tratamento farmacológico , Toxinas Botulínicas Tipo A/uso terapêutico , Distúrbios Distônicos/tratamento farmacológico , Espasticidade Muscular/tratamento farmacológico , Torcicolo/tratamento farmacológico , Idoso , Blefarospasmo/sangue , Estudos Transversais , Distúrbios Distônicos/sangue , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Espasticidade Muscular/sangue , Análise de Regressão , Estatísticas não Paramétricas , Torcicolo/sangue
14.
J Neural Transm (Vienna) ; 125(9): 1351-1354, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-29946929

RESUMO

Botulinum toxin (BT) is provided by several manufacturers producing a number of different drugs. Their potency is given in internationally standardised mouse units (MU). Clinical practise, however, reveals that the potency labelling of different BT drugs may not be identical. We wanted to use the mouse diaphragm assay (MDA) to compare the two BT drugs onabotulinumtoxinA (ONA) and incobotulinumtoxinA (INCO). For this, we measured the paresis time (PT) of different ONA or INCO doses. All BT came from several different and unexpired drug batches. PT for 20MU were 169.7 ± 28.9 min (ONA) and 132.3 ± 1.5 min (INCO) (p = 0.089), for 60MU 105.3 ± 10.1 min and 84.7 ± 4.2 min (p = 0.031), for 100MU 69.7 ± 1.5 min and 66.0 ± 7.0 min (p = 0.462) and for 140MU 74.7 ± 0.6 min and 62.3 ± 2.1 min (p = 0.100), respectively. The overall PT were 104.8 ± 12.5 and 86.3 ± 8.5 min (p = 0.178). Results presented here do not reveal differences in potency labelling between ONA and INCO, even when the full range of therapeutic doses are examined, although there was a trend towards stronger INCO effects. Data confirm previous reports on identical potency labelling of ONA and INCO. The MDA seems to be an appropriate instrument to test the potency labelling of other BT drugs as well, including new BT drugs currently under development.


Assuntos
Bioensaio , Toxinas Botulínicas Tipo A/farmacologia , Diafragma/efeitos dos fármacos , Fármacos Neuromusculares/farmacologia , Nervo Frênico/efeitos dos fármacos , Animais , Diafragma/inervação , Relação Dose-Resposta a Droga , Estimulação Elétrica , Contração Isométrica/efeitos dos fármacos , Camundongos
15.
J Neural Transm (Vienna) ; 124(10): 1223-1225, 2017 10.
Artigo em Inglês | MEDLINE | ID: mdl-28770389

RESUMO

Botulinum neurotoxin (BNT), the biologically active component of botulinum toxin (BT), is a large double-stranded protein susceptible to various physical and chemical influences. All BT type A (BT-A) drugs are stored as powders allowing shelf lives from 24 to 36 months. After reconstitution, the specified shelf life is reduced to 8-24 h. Some studies, however, suggest longer shelf life. We wanted to test the long-term stability of reconstituted BT-A drugs in the hemidiaphragm assay (HDA), a high quality BT potency test. For this incobotulinumtoxinA (INCO) was reconstituted and stored in at 4-8 °C, whilst at various points of time probes of it were taken and potency tested with the HDA. Altogether 18 measurements were performed throughout a period of 52.1 weeks. The paralysis time in the HDA was 67.3 ± 5.2 min (min. 59 min, max. 76 min). The linear regression line was described by y = -0.0163x + 67.582. The paralysis time measured during the first 10 weeks (n = 11) was 67.5 ± 5.3 min, during the last 10 weeks (n = 7) 67.1 ± 4.9 min. Reconstituted INCO does not show reduction of potency throughout 52 weeks as tested by the high quality HDA. Lack of complexing proteins does not de-stabilise INCO. Our data allow un-used reconstituted INCO to be stored for further use. This may have a considerable impact on the costs of BT therapy. Further studies will have to demonstrate sterility of the reconstituted BT drug beyond the so far reported 6 weeks.


Assuntos
Toxinas Botulínicas Tipo A , Diafragma/efeitos dos fármacos , Animais , Toxinas Botulínicas Tipo A/química , Toxinas Botulínicas Tipo A/economia , Toxinas Botulínicas Tipo A/farmacologia , Toxinas Botulínicas Tipo A/uso terapêutico , Composição de Medicamentos , Humanos , Fatores de Tempo
16.
Expert Rev Neurother ; 17(5): 487-494, 2017 05.
Artigo em Inglês | MEDLINE | ID: mdl-27852103

RESUMO

INTRODUCTION: Botulinum toxin (BT) is used in many medical specialties to treat muscle hyperactivity, exocrine gland hyperactivity and pain disorders. BT drugs consist of botulinum neurotoxin (BNT), complexing proteins (CP) and excipients. Antibodies can be formed against BNT and CP. When they are formed against BNT (BTAB) they can block BT's therapeutic efficacy thus producing antibody induced therapy failure (ABTF). Areas covered: BT applied and BTAB are in a functional balance within the body. ABTF is rare, but influences the treatment algorithms of BT therapy considerably. ABTF risk factors include BT doses given, interinjection intervals, booster injections and immunological quality of the BT drug. Testing for BTAB and interpretation of ABTF is complicated. As management of ABTF is frustrating, prevention of ABTF is of major importance. Improved antigenicity of new BT drugs may improve treatment algorithms of BT therapy, substandard antigenicity may very likely be their end. Expert commentary: Concern about ABTF has influenced the treatment algorithms of BT therapy considerably. Better understanding of ABTF may improve them and, thus, the outcome of BT therapy. New BT drugs may have further improved antigenicity, especially when their CP are removed. They may, however, fail because of antigenicity problems.


Assuntos
Toxinas Botulínicas/imunologia , Toxinas Botulínicas/uso terapêutico , Agitação Psicomotora/imunologia , Transtornos Somatoformes/imunologia , Algoritmos , Anticorpos Antibacterianos/biossíntese , Anticorpos Antibacterianos/imunologia , Clostridium botulinum/imunologia , Humanos , Agitação Psicomotora/tratamento farmacológico , Fatores de Risco , Transtornos Somatoformes/tratamento farmacológico
17.
J Neural Transm (Vienna) ; 123(5): 523-5, 2016 05.
Artigo em Inglês | MEDLINE | ID: mdl-27100914

RESUMO

Most botulinum toxin (BT) drugs are stored as powders which need to be reconstituted with normal saline before clinical use. As botulinum neurotoxin (BNT), the therapeutically active ingredient, is a large double-stranded protein the process of reconstitution should be performed with special attention to mechanical stress applied. We wanted to test the mechanical stability of BNT during the reconstitution process. For this, 100 MU onabotulinumtoxinA (Botox(®), Irvine, CA, USA) was reconstituted with 2.0 ml of NaCl/H2O. Gentle reconstitution (GR) was performed with a 5 ml syringe, a 0.90 × 70 mm injection needle, one cycle of injection-aspiration-injection and two gentle shakes of the vial. Aggressive reconstitution (AR) was performed with a 5 ml syringe, a 0.40 × 40 mm injection needle, ten injection-aspiration-injection cycles and 30 s of continuous shaking of the vial. AR increased the time to paralysis in the mouse hemidiaphragm assay (HDA) from 72.0 ± 4.6 to 106.0 ± 16.0 min (*p = 0.002, two-tailed t test after Kolmogorov-Smirnova test with Lilliefors correction for normal distribution). Construction of a calibration curve revealed that the increase in the time to paralysis was correlated with a loss of potency of from 100 to 58 MU (-42 %). BT users should use large diameter injection needles for reconstitution, apply two or three injection-aspiration-injection cycles and, maybe, shake the vials a few times to rinse the entire glass wall. Aggressive reconstitution with small diameter needles, prolonged injection-aspiration-injection and violent shaking should be avoided.


Assuntos
Toxinas Botulínicas/química , Toxinas Botulínicas/farmacologia , Diafragma/efeitos dos fármacos , Embalagem de Medicamentos , Fármacos Neuromusculares/farmacologia , Animais , Toxinas Botulínicas/normas , Relação Dose-Resposta a Droga , Camundongos , Fármacos Neuromusculares/química , Fármacos Neuromusculares/normas , Estatísticas não Paramétricas
18.
J Neural Transm (Vienna) ; 123(5): 527-31, 2016 05.
Artigo em Inglês | MEDLINE | ID: mdl-27002815

RESUMO

Botulinum toxin (BT) is injected intramuscularily and may produce injection site pain (ISP). We wanted to explore whether the pH value of the reconstituted BT drug contributes to ISP and, if so, what strategies can be applied to reduce it. In part 1 of the study, pH values of different reconstitution solutions and of major BT drugs reconstituted with different reconstitution solutions were assessed. In part 2, the effects of reconstitution with normal saline (NS) and Ringer acetate (RA) were compared intraindividually and in a double blind fashion in 34 patients with blepharospasm. pH values of reconstitution solutions were 5.52 ± 0.02 for NS, 6.98 for RA, 6.31 for Ringer lactate, 6.56 for electrolyte and 5.31 for bacteriostatic solution. pH values for NS-reconstitution were 6.09 ± 0.20 for Botox(®), 5.95 ± 0.24 for Dysport(®) and 5.81 ± 0.18 for Xeomin(®). pH values for RA-reconstitution were 6.95 ± 0.03 for Botox(®), 7.01 ± 0.02 for Dysport(®) and 6.87 ± 0.06 for Xeomin(®). By using RA instead of NS the pH could be increased by 0.86 for Botox(®), by 1.06 for Dysport(®) and by 1.06 for Xeomin(®). 47 % of the patients experienced less ISP when Botox(®)-RA was given rather than Botox(®)-NS, 76 % when Xeomin(®)-RA was given rather than Xeomin(®)-NS. None of the patients reported a difference in efficacy between NS- and RA-reconstitution. Despite previous reports, reconstituted BT type A drugs show acidic pH values. Normalising these pH values by use of RA instead of NS reduces ISP considerably without sacrificing clincial efficacy.


Assuntos
Toxinas Botulínicas/química , Toxinas Botulínicas/uso terapêutico , Composição de Medicamentos , Concentração de Íons de Hidrogênio , Fármacos Neuromusculares/uso terapêutico , Dor/tratamento farmacológico , Idoso , Blefarospasmo/tratamento farmacológico , Relação Dose-Resposta a Droga , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Fármacos Neuromusculares/química , Dor/etiologia , Fatores de Tempo
19.
Toxins (Basel) ; 7(12): 4895-905, 2015 Nov 25.
Artigo em Inglês | MEDLINE | ID: mdl-26610569

RESUMO

The historical method for the detection of botulinum neurotoxin (BoNT) is represented by the mouse bioassay (MBA) measuring the animal survival rate. Since the endpoint of the MBA is the death of the mice due to paralysis of the respiratory muscle, an ex vivo animal replacement method, called mouse phrenic nerve (MPN) assay, employs the isolated N. phrenicus-hemidiaphragm tissue. Here, BoNT causes a dose-dependent characteristic decrease of the contraction amplitude of the indirectly stimulated muscle. Within the EQuATox BoNT proficiency 13 test samples were analysed using the MPN assay by serial dilution to a bath concentration resulting in a paralysis time within the range of calibration curves generated with BoNT/A, B and E standards, respectively. For serotype identification the diluted samples were pre-incubated with polyclonal anti-BoNT/A, B or E antitoxin or a combination of each. All 13 samples were qualitatively correctly identified thereby delivering superior results compared to single in vitro methods like LFA, ELISA and LC-MS/MS. Having characterized the BoNT serotype, the final bath concentrations were calculated using the calibration curves and then multiplied by the respective dilution factor to obtain the sample concentration. Depending on the source of the BoNT standards used, the quantitation of ten BoNT/A containing samples delivered a mean z-score of 7 and of three BoNT/B or BoNT/E containing samples z-scores <2, respectively.


Assuntos
Toxinas Botulínicas Tipo A/toxicidade , Toxinas Botulínicas/toxicidade , Neurotoxinas/toxicidade , Nervo Frênico/efeitos dos fármacos , Paralisia Respiratória/induzido quimicamente , Animais , Bioensaio , Toxinas Botulínicas/análise , Toxinas Botulínicas Tipo A/análise , Soluções Tampão , Masculino , Carne/análise , Camundongos , Leite/química , Neurotoxinas/análise , Nervo Frênico/fisiopatologia , Paralisia Respiratória/fisiopatologia , Soro/química , Soroalbumina Bovina/química
20.
Toxicology ; 328: 48-56, 2015 Feb 03.
Artigo em Inglês | MEDLINE | ID: mdl-25497110

RESUMO

Clostridium difficile toxins A and B (TcdA and TcdB) belong to the class of large clostridial cytotoxins and inactivate by glucosylation some low molecular mass GTPases of the Rho-family (predominantly Rho, Rac and Cdc42), known as regulators of the actin cytoskeleton. TcdA and B also represent the main virulence factors of the anaerobic gram-positive bacterium that is the causal agent of pseudomembranous colitis. In our study, TcdB was chosen instead of TcdA for the well-known higher cytotoxic potency. Inactivation of Rho-family GTPases by this toxin in our experimental conditions induced morphological changes and reduction of electron-dense mast cell-specific granules in human mast cell line-1 (HMC-1) cells, but not cell death or permeabilisation of plasma-membranes. Previously reported patch-clamp dialysis experiments revealed that high intracellular free-Ca(2+) and GTPγS concentrations are capable of inducing exocytosis as indicated by significant membrane capacitance (Cm) increases in HMC-1 cells. In this study, we investigated the direct effects of TcdB upon HMC-1 cell "stimulated" Cm increase, as well as on "constitutive" secretion of hexosaminidase and interleukin-16 (IL-16). Compared to untreated control cells, HMC-1 cells incubated with TcdB for 3-24h exhibited a significant reduction of the mean absolute and relative Cm increase in response to free-Ca(2+) and GTPγS suggesting an inhibition of secretory processes by TcdB. In conclusion, the HMC-1 cell line represents a suitable model for the study of direct effects of C. difficile toxins on human mast cell secretory activity.


Assuntos
Proteínas de Bactérias/farmacologia , Toxinas Bacterianas/farmacologia , Cálcio/metabolismo , Exocitose/efeitos dos fármacos , Guanosina 5'-O-(3-Tiotrifosfato)/farmacologia , Hexosaminidases/metabolismo , Interleucina-16/metabolismo , Mastócitos/efeitos dos fármacos , Linhagem Celular , Diálise , Capacitância Elétrica , Humanos , Mastócitos/metabolismo , Potenciais da Membrana , Técnicas de Patch-Clamp , Fatores de Tempo
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