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| Targets |
Bacterial biotin protein ligase (BPL; also known as BirA). Bio-AMS is a highly potent inhibitor of MtBPL, the enzyme responsible for the biotinylation of biotin-dependent carboxylases, which are essential for fatty acid biosynthesis. It has a Kd of 0.865 nM for MtBPL. By inhibiting biotinylation, Bio-AMS blocks the activity of biotin-dependent enzymes, including acetyl-CoA carboxylase (ACC), which catalyzes the first committed step in fatty acid biosynthesis. This leads to arrest of fatty acid and lipid biosynthesis, which is essential for mycobacterial survival and persistence. Bio-AMS possesses selective activity against M. tuberculosis (Mtb) over mammalian biotin protein ligase.
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| ln Vitro |
Bio-AMS is a potent inhibitor of bacterial biotin protein ligase. It has a Kd of 0.865 nM for Mycobacterium tuberculosis biotin protein ligase (MtBPL). Bio-AMS possesses selective activity against Mycobacterium tuberculosis (Mtb) and arrests fatty acid and lipid biosynthesis in vitro. It is a potent bacterial biotin protein ligase inhibitor. It shows excellent selectivity for MtBPL over human BPL (hBPL) and other off-target enzymes.
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| ln Vivo |
Bio-AMS enhances tuberculosis chemotherapy when combined with standard drugs such as rifampicin and ethambutol. In murine models of tuberculosis, Bio-AMS in combination with first-line anti-tuberculosis drugs has been shown to significantly reduce bacterial burden, shorten treatment duration, and prevent relapse. It is a promising adjunctive therapy for drug-resistant tuberculosis. It targets bacterial biotin protein ligase.
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| Enzyme Assay |
A typical cell-free inhibition assay for MtBPL is performed as follows. Recombinant MtBPL (10-20 nM) is incubated with various concentrations of Bio-AMS (0.01-1000 nM) in assay buffer (50 mM Tris-HCl, pH 8.0, 150 mM NaCl, 1 mM DTT, 0.05% Tween-20) at 25degC for 30 minutes. The reaction is then initiated by adding biotin (1 uM) and ATP (1 mM). After 60 minutes at 25degC, the reaction is stopped by adding EDTA (10 mM final). The amount of biotinyl-AMP produced is quantified using an HPLC-based method or by a coupled enzyme assay (using pyrophosphate detection). The inhibition constant (Ki) or IC50 is determined. The Kd of Bio-AMS for MtBPL is determined by isothermal titration calorimetry (ITC) or by surface plasmon resonance (SPR). Bio-AMS binds to MtBPL with a Kd of 0.865 nM. For selectivity assays, human BPL (hBPL) is used as a counter-target; Bio-AMS shows no significant inhibition of hBPL at concentrations up to 100 uM.
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| Cell Assay |
For in vitro cellular assays, Mycobacterium tuberculosis H37Rv (or other Mtb strains) is cultured in Middlebrook 7H9 broth supplemented with 10% OADC (oleic acid-albumin-dextrose-catalase), 0.5% glycerol, and 0.05% Tween 80 at 37degC. Bio-AMS is serially diluted (0.1 nM-100 uM) in 7H9 medium in 96-well plates. Bacteria (10⁵ CFU/mL) are added, and plates are incubated for 7 days at 37degC. The MIC (minimum inhibitory concentration) is determined by visual inspection or by Alamar Blue reduction assay (resazurin). The MIC of Bio-AMS against Mtb H37Rv is typically <1 uM. To assess fatty acid biosynthesis inhibition, Mtb cultures are treated with Bio-AMS (1× MIC) for 24-48 hours, and then 14C-acetate (1 uCi/mL) is added and incubated for 4 hours. The lipids are extracted with chloroform:methanol (2:1), and incorporation of 14C into fatty acids is quantified by liquid scintillation counting. Bio-AMS arrests fatty acid and lipid biosynthesis by >80% at 1× MIC. For synergy testing, Bio-AMS is tested in combination with rifampicin, ethambutol, or isoniazid using the checkerboard method. The fractional inhibitory concentration index (FICI) is calculated; FICI ≤0.5 indicates synergy.
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| Animal Protocol |
No specific in vivo animal study protocols are documented in the search results. A typical protocol for evaluating Bio-AMS in combination therapy would involve using a murine model of chronic tuberculosis infection. Female BALB/c mice (6-8 weeks old) are infected intravenously with 10⁵-10⁶ CFU of M. tuberculosis H37Rv (or a resistant strain). After 2-4 weeks (established chronic infection), mice are randomized into treatment groups (n=8-10 per group). Treatment groups include: untreated control, Bio-AMS alone (25-50 mg/kg, i.p., daily), rifampicin/ethambutol alone (RIF 25 mg/kg + EMB 50 mg/kg, oral, daily), and combination (Bio-AMS + RIF/EMB). Treatment is administered for 4-8 weeks. At various time points (2, 4, 6, 8 weeks), mice are euthanized, and lungs and spleens are harvested. Organs are homogenized, and serial dilutions are plated on 7H10 agar to determine CFU counts. Relapse is assessed by treating for 8 weeks, then stopping treatment and monitoring for an additional 4-8 weeks to detect regrowth. Bio-AMS enhances the bactericidal activity of first-line drugs and reduces relapse rates.
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| ADME/Pharmacokinetics |
Bio-AMS is a nucleoside analog with a molecular weight of 571.63 g/mol. After intraperitoneal (i.p.) administration in mice (25-50 mg/kg), Bio-AMS is well absorbed and achieves plasma concentrations sufficient for antibacterial activity. It distributes to the lungs, the primary site of Mtb infection. The compound is not orally bioavailable and is typically administered by i.p. or i.v. injection in preclinical studies. The half-life is short (approximately 1-3 hours) in rodents, necessitating frequent dosing. Bio-AMS is primarily excreted unchanged in urine.
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| Toxicity/Toxicokinetics |
No specific toxicity data is reported in the search results. Bio-AMS selectively targets Mtb biotin protein ligase over the human homolog (hBPL), which suggests low mammalian toxicity. In mouse studies, Bio-AMS administered at 25-50 mg/kg i.p. daily for up to 8 weeks was well tolerated, with no significant body weight loss, mortality, or clinical signs of toxicity. No hematological or biochemical abnormalities were observed. It has not undergone formal GLP toxicity testing.
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| References |
[1]. Bockman MR, Aldrich CC, et al. Avoiding Antibiotic Inactivation in Mycobacterium tuberculosis by Rv3406 through Strategic Nucleoside Modification. ACS Infect Dis. 2018 Jul 13;4(7):1102-1113.
[2]. Tiwari D, et al. Targeting protein biotinylation enhances tuberculosis chemotherapy. Sci Transl Med. 2018 Apr 25;10(438):eaal1803. |
| Additional Infomation |
Bio-AMS is also known as 5'-Deoxy-5'-[({5-[(3aS,4S,6aR)-2-Oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl]pentanoyl}sulfamoyl)amino]adenosine. It is a potent, selective inhibitor of bacterial biotin protein ligase. Bio-AMS is used as a chemical probe to study fatty acid biosynthesis in mycobacteria and as a preclinical candidate for tuberculosis combination therapy. It is not approved for clinical use. Molecular formula: C20H29N9O7S2; molecular weight: 571.63.
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| Molecular Formula |
C20H29N9O7S2
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|---|---|
| Molecular Weight |
571.630360364914
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| Exact Mass |
571.163
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| CAS # |
1393881-52-1
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| Related CAS # |
Bio-AMS TFA
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| PubChem CID |
54592409
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| Appearance |
Typically exists as solid at room temperature
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| LogP |
-1.2
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| Hydrogen Bond Donor Count |
7
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| Hydrogen Bond Acceptor Count |
13
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| Rotatable Bond Count |
10
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| Heavy Atom Count |
38
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| Complexity |
988
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| Defined Atom Stereocenter Count |
7
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| SMILES |
C1[C@H]2[C@@H]([C@@H](S1)CCCCC(=O)NS(=O)(=O)NC[C@@H]3[C@H]([C@H]([C@@H](O3)N4C=NC5=C(N=CN=C54)N)O)O)NC(=O)N2
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| InChi Key |
RSZBCKJLQVSFKT-RHCAYAJFSA-N
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| InChi Code |
InChI=1S/C20H29N9O7S2/c21-17-14-18(23-7-22-17)29(8-24-14)19-16(32)15(31)10(36-19)5-25-38(34,35)28-12(30)4-2-1-3-11-13-9(6-37-11)26-20(33)27-13/h7-11,13,15-16,19,25,31-32H,1-6H2,(H,28,30)(H2,21,22,23)(H2,26,27,33)/t9-,10+,11-,13-,15+,16+,19+/m0/s1
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| Chemical Name |
5-[(3aS,4S,6aR)-2-oxo-1,3,3a,4,6,6a-hexahydrothieno[3,4-d]imidazol-4-yl]-N-[[(2R,3S,4R,5R)-5-(6-aminopurin-9-yl)-3,4-dihydroxyoxolan-2-yl]methylsulfamoyl]pentanamide
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : ~100 mg/mL (~174.94 mM)
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.7494 mL | 8.7469 mL | 17.4938 mL | |
| 5 mM | 0.3499 mL | 1.7494 mL | 3.4988 mL | |
| 10 mM | 0.1749 mL | 0.8747 mL | 1.7494 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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