We aimed to evaluate the activity of PBTZ169 and pretomanid against non-tuberculous mycobacteriosis (NTM) in vitro and in vivo.
The minimum inhibitory concentrations (MICs) of 11 antibiotics, against slow-growing mycobacteria (SGMs) and rapid-growing mycobacteria (RGMs) were tested using the microplate alamarBlue assay. The in vivo activities of bedaquiline, clofazimine, moxifloxacin, rifabutin, PBTZ169 and pretomanid against four common NTMs were assessed in murine models.
PBTZ169 and pretomanid had MICs of >32 μg/mL against most NTM reference and clinical strains. However, PBTZ169 was bactericidal against Mycobacterium abscessus (3.33 and 1.49 log10 CFU reductions in the lungs and spleen, respectively) and Mycobacterium chelonae (2.29 and 2.24 CFU reductions in the lungs and spleen, respectively) in mice, and bacteriostatic against Mycobacterium avium and Mycobacterium fortuitum. Pretomanid dramatically decreased the CFU counts of M. abscessus (3.12 and 2.30 log10 CFU reductions in the lungs and spleen, respectively), whereas it showed moderate inhibition of M. chelonae and M. fortuitum. Bedaquiline, clofazimine, and moxifloxacin showed good activities against four NTMs in vitro and in vivo. Rifabutin did not inhibit M. avium and M. abscessus in mice.
PBTZ169 appears to be a candidate for treating four common NTM infections. Pretomanid was more active against M. abscessus, M. chelonae and M. fortuitum than against M. avium.

暂无摘要。

Tuberculosis, and especially multidrug-resistant tuberculosis (MDR-TB), is a major global health threat which emphasizes the need to develop new agents to improve and shorten treatment of this difficult-to-manage infectious disease. Among the new agents, macozinone (PBTZ169) is one of the most promising candidates, showing extraordinary potency in vitro and in murine models against drug-susceptible and drug-resistant Mycobacterium tuberculosis. A previous analytical method using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) was developed by our group to support phase I clinical trials of PBTZ169. These plasma sample analyses revealed the presence of several additional metabolites among which the most prominent was H2PBTZ, a reduced species obtained by dearomatization of macozinone, one of the first examples of Meisenheimer Complex (MC) metabolites identified in mammals. Identification of these new metabolites required the optimization of our original method for enhancing the selectivity between isobaric metabolites as well as for ensuring optimal stability for H2PBTZ analyses. Sample preparation methods were also developed for plasma and urine, followed by extensive quantitative validation in accordance with international bioanalytical method recommendations, which include selectivity, linearity, qualitative and quantitative matrix effect, trueness, precision and the establishment of accuracy profiles using β-expectation tolerance intervals for known and newer analytes. The newly optimized methods have been applied in a subsequent Phase Ib clinical trial conducted in our University Hospital with healthy subjects. H2PBTZ was found to be the most abundant species circulating in plasma, underscoring the importance of measuring accurately and precisely this unprecedented metabolite. Low concentrations were found in urine for all monitored analytes, suggesting extensive metabolism before renal excretion.

Macozinone (MCZ; PBTZ169) is a first-in-class antituberculosis clinical-stage benzothiazinone-based drug candidate. Although its efficacy and safety have been strongly proven in several preclinical and clinical studies, the physicochemical and pharmacokinetic properties specific to MCZ required further optimization. Accordingly, this study aimed to evaluate the pharmacokinetics of MCZ administered as extended-release (ER) tablets F2 and F6 compared to immediate-release (IR) dispersible tablets for oral suspension. Oral absorption of MCZ from ER tablets was significantly different from that of IR tablets after a single oral dose in Beagle dogs in both fasted and fed states. In addition, food directly affects the bioavailability of MCZ from ER tablets but does not affect it from IR tablets. The high values of relative bioavailability of the prolonged-release tablets F2 and F6 compared to the IR tablets may indicate an indirect confirmation of their gastroretentive properties. Taken together, pharmacokinetic parameters have demonstrated that these MCZ oral formulations not just enhance drug bioavailability but may also improve regimen adherence by reducing MCZ dose frequency and reducing the development of drug resistance. IMPORTANCE Macozinone (MCZ) is the newest first-in-class clinical-stage benzothiazinone-based drug candidate for the treatment of tuberculosis. Yet, the extremely low oral bioavailability of MCZ, a major problem in clinical trials, needed to be addressed, and we are pleased to present our attempts to solve this issue. We report that extended-release tablets of MCZ significantly increased key pharmacokinetic parameters in the preclinical setting. We suggest that these MCZ oral formulations not just enhance drug bioavailability but may also improve regimen adherence by reducing MCZ dose frequency and reducing the development of drug resistance.

Decaprenyl-phosphoryl-ribose 2\'-epimerase (DprE1) inhibitors are an innovative and futuristic orally active group of antituberculosis agents. A few DprE1 inhibitors are in the clinical trial for tuberculosis (TB), including macozinone. This review highlights the discovery, developmental status, clinical studies, patents, and prospects of macozinone (MCZ). The patent and non-patent literature search was done by entering keywords such as macozinone; MCZ; PBTZ169; PBTZ-169 in Pubmed, Espacenet, Patentscope, and the USPTO databases. However, data on Sci-Finder was searched using CAS registry number: 1377239-83-2. MCZ clinical trial studies were retrieved from the clinicaltrials.gov database using the exact keywords. The chemical structure of MCZ was disclosed in 2009. Accordingly, patents/patent applications published from 2009 to June 12, 2022, have been discussed herein. MCZ and MCZ hydrochloride salt patents were granted in 2014 and 2019, respectively, in the USA. The patent literature and the clinical trial studies suggest capsule, tablet, and suspension formulations of crystalline MCZ and its hydrochloride salt as the possible and prospective dosage forms to treat TB. Some combinations of MCZ with other drugs (chloroquine, telacebec, tafenoquine, TBI-166, and sanfetrinem) with improved anti-TB efficacy have been documented. Based on the literature covered in this review article on the clinical studies and patents applied/granted to MCZ, it can be inferred that MCZ seems to be a promising DprE1 inhibitor and could help to tackle the emerging dilemma of drug-resistant either as a monotherapy or in combination with additional anti-TB agents. Furthermore, the authors anticipate the development of new combinations, salts, and polymorphs of MCZ as anti-TB agents shortly. This review article might prove beneficial to the scientific community as it summarizes chemistry, pharmacology and provides an update on the clinical studies and patents/patent applications of one of the emerging anti-TB drugs in one place.

BACKGROUND: Tuberculosis (TB) is one of the top ten causes of death worldwide. Improvement of the treatment options via development of new drugs and treatment regimens that would be more convenient for patients is one of key options of improving the effecacy of the TB prevention and careis. Since the creation of new treatment regimens by minimizing the number of the drugs used and reducing the duration of treatment is the most promising and correct direction, macozinone, a new candidate of the benzothiazinone series, can become the basis for development of new chemotherapy regimens for drug-resistant forms of TB including the combination of macozinone with the most effective modern anti-TB drugs.
OBJECTIVE: Comparative evaluation of the pharmacokinetic properties of macozinone capsules 80 mg and the new dosage form a dispersible tablet for preparation of oral solution.
METHODS: Solubility of the substance macozinone in biorelevant media in vitro, permeability of macozinone in the test Caco-2 in vitro, as well as pharmacokinetics of macozinone in dogs in vivo were evaluated.
RESULTS: The solubility assessment in biorelevant media showed that the average limit of macozinone substance dissolution in the pH 5.0 acetate buffer solution was from 6 to 9 mg/l, in FaSSIF medium (fasted) from 2.5 to 4 mg/l, and in FeSSIF medium (after meals) from 16.8 to 29 mg/l. It is established that the cell permeability of the pharmaceutical substance macozinone in the CACO-2 test system is on average 2.510-6cm/s in the forward direction from the apical to basolateral cell membrane, and 1.510-6cm/s in the reverse direction, which corresponds to low permeability. The main pharmacokinetic parameters of macozinone dispersable tablets 160 mg, after dosing with food and on an empty stomach, as well as capsules 80 mg, when administered on an empty stomach in vivo studies in dogs are presented.
CONCLUSIONS: The specific physicochemical properties of macozinone, the problems of developing the new dosage form, as well as ways of solving some of them are presented.
CONCLUSIONS: In the process of new dosage forms development, the existing chemical properties of the macozinone substance should be considered. One of the promising ways of increasing bioavailability and, consiquently, efficacy is development a fundamentally new drug form with modified release within the absorption window.
Введение.Туберкулез (ТБ) входит в десятку наиболее распространенных причин смертности в мире. Одной из ключевых задач повышения эффективности борьбы с ТБ является поиск новых лекарственных препаратов и схем лечения, которые наиболее удобны для пациентов и, при этом, достаточно эффективны. Поскольку создание новых режимов терапии с минимизацией количества используемых препаратов и сокращением длительности сроков лечения наиболее перспективное направление, макозинон, новый кандидат бензотиазинонового ряда, может стать основой для создания новых режимов химиотерапии лекарственно-устойчивых форм ТБ, в том числе и в комбинации с наиболее эффективными на сегодняшний день противотуберкулезными лекарственными средствами. Цель.Сравнительная оценка фармакокинетических свойств лекарственной формы макозинона капсулы 80 мг и новой лекарственной формы диспергируемая таблетка для приготовления раствора для приема внутрь 320 мг. Материалы и методы.Проводились оценка растворимости макозинона в биорелевантных средахin vitro, изучение проницаемости макозинона в тесте Caco-2in vitro, а также исследование фармакокинетики на собакахin vivo. Результаты.Оценка растворимости в биорелевантных средах показала, что в среднем предел растворения субстанции макозинон в среде ацетатного буферного раствора рН 5,0 составил от 6 до 9 мг/л, в среде FaSSIF (натощак) от 2,5 до 4 мг/л, а в среде FeSSIF (после еды) от 16,8 до 29 мг/л. Также установлено, что клеточная проницаемость фармацевтической субстанции макозинон в тест-системе Сасо-2 составляет в среднем 2,510-6cм/с в прямом направлении от апикальной к базолатеральной мембране клеток и 1,510-6cм/с в обратном направлении, что соответствует низкой проницаемости. Представлены основные фармакокинетические параметры макозинона после дозирования диспергируемой таблетки на фоне приема пищи и натощак, а также капсул 80 мг при введении их натощак собакам. Обсуждение.В данной работе описаны специфические физико-химические свойства макозинона, проблемы, с которыми пришлось столкнуться в процессе разработки лекарственной формы препарата, а также способы решения некоторых из них. Заключение.При разработке перспективных для применения в клинической практике лекарственных форм макозинона необходимо учитывать существующие химические свойства субстанции макозинон. Одним из перспективных направлений решения повышения биодоступности, а следовательно, и эффективности является создание принципиально новой лекарственной формы с модифицированным высвобождением, учитывающей узкое окно абсорбции макозинона.

8-Nitro-1,3-benzo-thia-zin-4-ones are a promising class of new anti-tubercular agents, two candidates of which, namely BTZ043 and PBTZ169 (INN: macozinone), have reached clinical trials. The crystal and mol-ecular structures of two synthetic precursors, 2-chloro-3-nitro-5-(tri-fluoro-meth-yl)benzoic acid, C8H3ClF3NO4 (1), and 2-chloro-3-nitro-5-(tri-fluoro-meth-yl)benzamide, C8H4ClF3N2O3 (2), are reported. In 1 and 2, the respective carb-oxy, carboxamide and the nitro groups are significantly twisted out of the plane of the benzene ring. In 1, the nitro group is oriented almost perpendicular to the benzene ring plane. In the crystal, 1 and 2 form O-H⋯O and N-H⋯O hydrogen-bonded dimers, respectively, which in 2 extend into primary amide tapes along the [101] direction. The tri-fluoro-methyl group in 2 exhibits rotational disorder with an occupancy ratio of 0.876 (3):0.124 (3).

Tuberculosis is a chronic infectious disease, usually localized in the respiratory system and representing one of the most important global social and biomedical health problems associated with the spread of therapy-resistant forms (multidrug-resistant and extensively drug-resistant tuberculosis). One of the most promising targets for the development of antimycobacterial drugs is the enzyme DprE1, which is involved in the synthesis of the cell wall of mycobacteria. In the series of DprE1 inhibitor drugs, the most advanced drug is PBTZ169 (INN maсozinone). Clinical trials (CT) of the efficacy and safety of macozinone are conducted by the pharmaceutical company LLC NEARMEDIC PLUS in the Russian Federation, and in other countries (Sponsors: Innovative Medicines for Tuberculosis Foundation, cole polytechnique fdrale de Lausanne and Bill and Melinda Gates Foundation). The publication describes results of completed I, IIa and Ib phases CT, conducted in the Russian Federation.
OBJECTIVE: The goal of phase I CT was to assess the safety, tolerability and pharmacokinetics (PK) of PBTZ169, 40 mg capsule, after single and multiple administration under fasting conditions in increasing doses in healthy volunteers. The goal of phase IIa CT was to study the efficacy (in terms of early bactericidal activity EBA), safety and PK of the drug PBTZ169, 80 mg capsules, in various doses, when used as monotherapy in patients with newly diagnosed respiratory tuberculosis with bacterial excretion and retained sensitivity to isoniazid and rifampicin. The purpose of phase Ib CT was to evaluate the safety, tolerability, PK of PBTZ169, 80 mg capsule, after single, double and multiple administration under fasting conditions in increasing doses, as well as the effect of food on its bioavailability in healthy volunteers.
METHODS: The data of 100 healthy volunteers and 15 patients with newly diagnosed pulmonary tuberculosis, who received the study medication PBTZ169, capsules 40 mg and 80 mg, in the dose range 40 mg 1280 mg of PBTZ169, obtained during phase I, IIa and Ib CTs were analyzed. During I phases CTs, safety, tolerability, and PK of the drug after a single and multiple administration under fasting condition and after meals at rising doses were evaluated. The safety assessment included evaluation of AE/SAE, vital signs, ECG results, and laboratory tests results in the safety population. In the course of phase IIa CT, in addition to safety, tolerance, and PK evaluation, the efficacy of the drug (in terms of EBA) using sputum culture on agar with CFU/ml counting (main method) and quantitative PCR method (auxiliary method) was evaluated.
RESULTS: During all CTs, a high safety and tolerability profile was shown, the main PK parameters of the drug and the efficacy were described. PBTZ169 demonstrated linear PK in the dosage range up to 640 mg after single and multiple administration, a statistically significant of EBA of the drug after monotherapy at the dose of 640 mg/day was demonstrate, and it was concluded that the preferred regimen of the drug PBTZ169 intake is administration after meals. Good tolerability and a favorable safety profile of the drug in the studied doses range were demonstrate during all the CTs.
CONCLUSIONS: One of the most promising and currently studied drugs-inhibitors of DprE1, a new target for the cell wall of mycobacteria, is PBTZ169 or macozinone, which is being develop by the Russian pharmaceutical company NEARMEDIC PLUS ltd.
Туберкулез хроническое инфекционное заболевание, как правило, локализующееся в органах дыхания и представляющее одну из важнейших глобальных социальных и медико-биологических проблем здравоохранения, связанную с распространением устойчивых к терапии форм (туберкулез с множественной и широкой лекарственной устойчивостью). Одной из наиболее перспективных мишеней для разработки антимикотических лекарственных препаратов является фермент DprE1, участвующий в синтезе клеточной стенки микобактерий. В ряду препаратов ингибиторов DprE1 наиболее изучен препарат PBTZ169 (международное непатентованное наименование макозинон), клинические исследования (КИ) эффективности и безопасности которого проводились как на территории Российской Федерации (спонсор ООО НИАРМЕДИК ПЛЮС), так и за рубежом (спонсоры Фонд Innovative Medicines for Tuberculosis, cole polytechnique fdrale de Lausanne и Bill and Melinda Gates Foundation). В публикации описаны результаты завершенных КИ I, IIа и Ib фаз, проведенных на территории Российской Федерации. Цель. Цель I фазы КИ заключалась в оценке безопасности, переносимости и фармакокинетики препарата PBTZ169, капсулы 40 мг, после однократного и многократного применения натощак в возрастающих дозах у здоровых добровольцев. Цель IIа фазы КИ заключалась в изучении эффективности (по параметру ранней бактерицидной активности РБА), безопасности и фармакокинетики препарата PBTZ169, капсулы 80 мг, в разных дозах при применении в качестве монотерапии у пациентов с впервые выявленным туберкулезом органов дыхания с бактериовыделением и сохраненной чувствительностью к изониазиду и рифампицину. Цель Ib фазы КИ заключалась в оценке безопасности, переносимости, фармакокинетики препарата PBTZ169, капсулы 80 мг, при однократном, двукратном и многократном применении натощак в возрастающих дозах, а также влияния пищи на его биодоступность у здоровых добровольцев. Материалы и методы. Проанализированы данные, полученные в результате I, IIа и Ib фаз КИ PBTZ169, в ходе которых 100 здоровых добровольцев и 15 пациентов с впервые выявленным туберкулезом легких принимали исследуемый препарат PBTZ169, капсулы 40 и 80 мг, в диапазоне доз от 40 до 1280 мг. В ходе КИ I фазы изучались безопасность, переносимость и фармакокинетика применяемых доз натощак и после еды при однократном и многократном применении. Оценка безопасности включала определение нежелательных явлений/серьезных нежелательных явлений, жизненно важных функций, данных электрокардиографии и лабораторных анализов всех добровольцев, принимавших исследуемый препарат. В ходе КИ IIа фазы помимо изучения безопасности, переносимости и фармакокинетики оценивали эффективность препарата по параметру РБА с использованием посева мокроты на агар с подсчетом КОЕ/мл (основной метод) и метода количественной полимеразной цепной реакции (вспомогательный метод). Результаты. В ходе всех КИ показан высокий профиль безопасности и переносимости, описаны основные фармакокинетические параметры препарата и эффективность по показателю РБА. Сделано заключение о тенденции линейности фармакокинетики в диапазоне дозировок до 640 мг при однократном и многократном приеме препарата PBTZ169, установлена статистически значимая РБА препарата после монотерапии в дозе 640 мг/сут, а также сделан вывод о предпочтительном режиме приема препарата PBTZ169 после приема пищи. Во всех КИ продемонстрированы хорошая переносимость и благоприятный профиль безопасности препарата в исследованном диапазоне доз. Заключение. Одним из наиболее перспективных и изученных на сегодняшний день препаратов ингибиторов DprE1, новой мишени клеточной стенки микобактерий, является PBTZ169 (макозинон), разработкой которого занимается российская фармацевтическая компания ООО НИАРМЕДИК ПЛЮС.

The causative agent of tuberculosis (TB), Mycobacterium tuberculosis and more recently totally drug-resistant strains of M. tuberculosis, display unique mechanisms to survive in the host. A four-drug treatment regimen was introduced 40 years ago but the emergence of multidrug-resistance and more recently TDR necessitates the identification of new targets and drugs for the cure of M. tuberculosis infection. The current efforts in the drug development process are insufficient to completely eradicate the TB epidemic. For almost five decades the TB drug development process remained stagnant. The last 10 years have made sudden progress giving some new and highly promising drugs including bedaquiline, delamanid, and pretomanid. Many of the candidates are repurposed compounds, which were developed to treat other infections but later, exhibited anti-TB properties also. Each class of drug has a specific target and a definite mode of action. These targets are either involved in cell wall biosynthesis, protein synthesis, DNA/RNA synthesis, or metabolism. This review discusses recent progress in the discovery of newly developed and Food and Drug Administration approved drugs as well as repurposed drugs, their targets, mode of action, drug-target interactions, and their structure-activity relationship.

In this study, we demonstrate that PBTZ169 exhibits significant differences in in vitro activity against multiple Mycobacterium species. The amino acid polymorphism at codon 387 of decaprenylphosphoryl-beta-d-ribose oxidase (DprE1) can be used as a surrogate marker for in vitro susceptibility to PBTZ169 in mycobacteria. In addition, the amino acid substitution at codon 154 in DprE1 may be associated with acquired resistance to PBTZ169 in the Mycobacterium fortuitum mutants.