| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| 50mg |
|
||
| 100mg | |||
| Other Sizes |
| Targets |
TCL-053 is a lipid-based delivery vehicle rather than a conventional enzyme inhibitor or receptor-targeting drug. Its primary biological target is the cellular membrane and the endosomal trafficking pathway. Following cellular uptake via endocytosis, the ionizable nature of TCL-053 (pKa = 6.8) facilitates endosomal escape through the proton sponge effect, releasing the nucleic acid payload into the cytosol. The lipid nanoparticles function by encapsulating therapeutic nucleic acids and mediating their intracellular delivery. TCL-053 does not bind to a specific protein target in the traditional sense; instead, it interacts with lipid bilayers and endosomal membranes to enable payload delivery. The targeting specificity is conferred by the LNP formulation and administration route rather than by TCL-053 itself. For muscle targeting, intramuscular injection or limb perfusion allows the LNPs to accumulate in skeletal muscle tissue. The lipid's ionizable property is crucial for its function—at physiological pH (7.4), it is relatively neutral, reducing nonspecific interactions and toxicity, while in the acidic endosomal environment (pH ~5-6), it becomes positively charged, promoting membrane destabilization and payload release into the cytoplasm.
|
|---|---|
| ln Vitro |
The dissociation constant for TCL053 is pKa = 6.8[1]. 79.1 nM in size, 96% encapsulation rate, and TCL053: DPPC: Cholesterol: DMG-PEG=60: 10.6: 27.3: 2.1 are the measurements [1].
In vitro characterization of TCL-053-based LNPs demonstrates excellent physicochemical properties for nucleic acid delivery. The optimized formulation with TCL053:DPPC:Cholesterol:DMG-PEG at a molar ratio of 60:10.6:27.3:2.1 achieves an encapsulation efficiency of 96% for mRNA payloads, with an average particle size of 79.1 nm as determined by dynamic light scattering. The lipid exhibits a dissociation constant pKa = 6.8, which is optimal for endosomal escape while maintaining stability in circulation. The LNPs efficiently encapsulate Cas9 mRNA and sgRNA, protecting the nucleic acids from degradation and facilitating cellular uptake. In cell culture studies, TCL053-LNPs demonstrate effective delivery of CRISPR components to target cells, resulting in successful gene editing. The encapsulation efficiency of 96% indicates that nearly all nucleic acid payload is protected within the LNP core. The small particle size (<100 nm) is favorable for cellular uptake via endocytosis and for extravasation into tissues. The polydispersity index and zeta potential of the formulations are consistent with stable LNP preparations suitable for in vivo applications. |
| ln Vivo |
Compared to other in vivo mRNA delivery reagents (in vivo-jetRNA), TCL053-based LNP-encapsulated Cas9 mRNA (TCL053-LNP-CRISPR) (20 µg total RNA; im) has a higher efficacy for exon skipping and genome editing [1]. LNP based on TCL053 has the ability to target various muscle groups by means of limb perfusion. It also possesses low immunogenicity and can be administered repeatedly [1]. Skeletal muscle disease research can make use of TCL053-based LNP, which is a CRISPR-Cas9 delivery vector [1]. In a DMD mouse model with humanized exon sequences, TCL053-based LNP induces stable genomic exon skipping and restores dystrophin protein [1].
In vivo studies demonstrate that TCL-053-based LNPs encapsulating Cas9 mRNA (TCL053-LNP-CRISPR) at a dose of 20 µg total RNA administered via intramuscular injection exhibit high efficacy for genome editing and exon skipping. In a Duchenne muscular dystrophy (DMD) mouse model harboring a humanized exon sequence, TCL053-LNP induces stable genomic exon skipping and restores dystrophin protein expression in skeletal muscle. The editing efficacy is superior to that of other in vivo mRNA delivery reagents such as in vivo-jetRNA. Importantly, TCL053-based LNPs can target multiple muscle groups through limb perfusion, a significant advantage for treating systemic muscle disorders. The LNPs demonstrate low immunogenicity, enabling repeated administrations without triggering neutralizing immune responses that would diminish efficacy. This is a critical feature for chronic conditions like DMD that require ongoing treatment. The lipid nanoparticles effectively deliver CRISPR-Cas9 components to muscle tissue, achieving therapeutic levels of gene editing. The restoration of dystrophin protein in the DMD mouse model confirms the functional relevance of the gene editing approach. The combination of high editing efficiency, low immunogenicity, and repeat dosing capability positions TCL053-LNP as a promising platform for in vivo gene therapy applications. |
| Enzyme Assay |
As TCL-053 is a lipid excipient rather than a drug targeting an enzyme or receptor, conventional enzyme/receptor binding assays are not applicable. Instead, physicochemical characterization methods are employed to evaluate the lipid's properties. The dissociation constant (pKa = 6.8) is determined by standard potentiometric titration, measuring the lipid's protonation behavior as a function of pH. This is critical for predicting endosomal escape capability. LNP formulation optimization involves particle size analysis using dynamic light scattering (DLS) to ensure uniform nanoparticle formation (typically <100 nm). Encapsulation efficiency is measured by fluorescent dye exclusion assays or gel electrophoresis, where free (unencapsulated) nucleic acids are separated from encapsulated ones, and the percentage of encapsulated payload is calculated. For the optimized TCL053 formulation, encapsulation efficiency of 96% and particle size of 79.1 nm have been reported. Lipid:payload ratios are optimized to achieve maximal encapsulation while maintaining nanoparticle stability. The molar ratio of TCL053:DPPC:Cholesterol:DMG-PEG = 60:10.6:27.3:2.1 represents the optimized formulation for skeletal muscle delivery.
|
| Cell Assay |
In vitro cell culture studies with TCL-053-based LNPs involve incubation of cells with LNP formulations encapsulating nucleic acid payloads such as Cas9 mRNA and sgRNA. Cells are typically treated with LNPs at various concentrations and incubation times to determine optimal delivery conditions. Gene editing efficiency is assessed by sequencing of target genomic loci or by functional assays such as reporter gene activation or protein expression analysis. For CRISPR-Cas9 delivery, cells are treated with TCL053-LNPs encapsulating both Cas9 mRNA and sgRNA targeting a specific gene locus. After incubation (typically 24-72 hours), cells are harvested for analysis of editing efficiency using T7 endonuclease I assay, Sanger sequencing, or next-generation sequencing. Protein expression changes (e.g., dystrophin restoration) are evaluated by Western blot or immunofluorescence. Cytotoxicity of the LNP formulation is assessed using standard assays such as MTT or CellTiter-Glo to ensure biocompatibility. The encapsulation rate of 96% and particle size of 79.1 nm have been measured for the optimized TCL053 formulation, confirming the quality of the LNP preparation before cell treatment.
|
| Animal Protocol |
In vivo animal experiments using TCL-053-based LNPs are conducted in mouse models. For Duchenne muscular dystrophy research, DMD mouse models with humanized exon sequences are used. TCL053-LNP-CRISPR (20 µg total RNA consisting of Cas9 mRNA and sgRNA) is administered via intramuscular injection into the gastrocnemius muscle or other target muscle groups. For broader muscle targeting, limb perfusion is employed, where the LNP solution is infused through the vascular system of a limb to achieve distribution to multiple muscle groups. After administration, tissues are harvested at various time points (typically 1-4 weeks post-injection) for analysis. Genomic DNA is extracted from muscle tissues and subjected to sequencing to quantify exon skipping and genome editing efficiency. Protein expression is assessed by Western blot and immunofluorescence staining of muscle sections to detect dystrophin restoration. The low immunogenicity of TCL053-LNPs is evaluated by measuring cytokine levels and antibody responses after repeated administrations. Histological analysis is performed to assess muscle morphology and any signs of tissue damage. The DMD mouse model studies demonstrate stable genomic exon skipping and dystrophin protein restoration, confirming the therapeutic potential of the approach.
|
| ADME/Pharmacokinetics |
As a lipid nanoparticle component, TCL-053 is not subject to conventional pharmacokinetic analysis of small molecules. Instead, the pharmacokinetic behavior of the LNP formulation is characterized by the circulation time, biodistribution, and clearance of the nanoparticles. TCL053-based LNPs exhibit extended circulation in the bloodstream, allowing accumulation in target tissues such as skeletal muscle. The ionizable nature of TCL-053 (pKa = 6.8) contributes to the favorable pharmacokinetic profile by reducing nonspecific protein adsorption and opsonization, which would otherwise lead to rapid clearance by the reticuloendothelial system. Upon intramuscular injection or limb perfusion, the LNPs localize to muscle tissue where they are taken up by cells via endocytosis. The low immunogenicity of TCL053-LNPs allows repeated administrations without accelerated clearance due to anti-PEG or anti-lipid antibody responses. The encapsulation efficiency of 96% ensures that the nucleic acid payload remains protected from nuclease degradation during circulation. The small particle size (79.1 nm) facilitates extravasation and tissue penetration. The lipid is metabolized through natural lipid degradation pathways, with clearance primarily via hepatic and renal routes.
|
| Toxicity/Toxicokinetics |
TCL-053-based LNPs demonstrate a low immunogenicity profile, which is a critical safety feature for therapeutic applications requiring repeated administration. Studies in mouse models show that repeated dosing does not elicit significant neutralizing immune responses against the LNP components, allowing sustained gene editing efficacy over multiple treatment cycles. This is in contrast to many other lipid delivery systems that trigger anti-PEG antibodies or inflammatory responses upon repeat dosing. No significant off-target toxicity or organ damage has been reported in the published studies. The LNPs are generally well-tolerated in mouse models at the tested doses (20 µg total RNA per injection). As with all lipid nanoparticle formulations, potential toxicities relate to the lipid components themselves, including complement activation-related pseudoallergy (CARPA) and hepatotoxicity at high doses, though these have not been reported for TCL053 at therapeutic doses. The product is for research use only and is not approved for human therapeutic use. Standard laboratory safety precautions should be followed when handling the compound. The favorable safety profile, combined with high editing efficiency, supports the continued development of TCL053-LNPs for gene therapy applications.
|
| References | |
| Additional Infomation |
TCL-053 is a next-generation ionizable lipid that represents a significant advancement in LNP technology for nucleic acid delivery. The key publication describing its application is Kenjo E, et al. "Low immunogenicity of LNP allows repeated administrations of CRISPR-Cas9 mRNA into skeletal muscle in mice," Nature Communications, 2021 Dec 8;12(1):7101. This study demonstrated that TCL053-LNPs could achieve stable genomic exon skipping and dystrophin restoration in a DMD mouse model. The lipid's low immunogenicity is a distinguishing feature that enables repeated administration, addressing a major limitation of many existing LNP platforms. TCL053 is not approved for clinical use and is intended for research purposes only, specifically for gene therapy and genome editing applications targeting skeletal muscle disorders. The compound is supplied as a liquid and should be stored at -20°C for up to 2 years in solution. Synonyms include (9Z,9'Z)-2-(((4-(dimethylamino)butanoyl)oxy)methyl)-2-(((Z)-tetradec-9-enoyloxy)methyl)propane-1,3-diyl bis(tetradec-9-enoate). The lipid is typically used in combination with DPPC, cholesterol, and DMG-PEG to form LNPs. The optimized molar ratio of TCL053:DPPC:Cholesterol:DMG-PEG = 60:10.6:27.3:2.1 yields particles of 79.1 nm with 96% encapsulation efficiency. The research community continues to explore TCL053 for various gene editing applications beyond DMD, including other neuromuscular disorders.
|
| Molecular Formula |
C53H95NO8
|
|---|---|
| Molecular Weight |
874.32
|
| Exact Mass |
873.705
|
| CAS # |
2361162-70-9
|
| PubChem CID |
154641272
|
| Appearance |
Colorless to light yellow liquid
|
| LogP |
16.2
|
| Hydrogen Bond Donor Count |
0
|
| Hydrogen Bond Acceptor Count |
9
|
| Rotatable Bond Count |
49
|
| Heavy Atom Count |
62
|
| Complexity |
1050
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
CCCC/C=C\CCCCCCCC(=O)OCC(COC(=O)CCCN(C)C)(COC(=O)CCCCCCC/C=C\CCCC)COC(=O)CCCCCCC/C=C\CCCC
|
| InChi Key |
MJPUTLHISFHJJZ-QMJZYSMNSA-N
|
| InChi Code |
InChI=1S/C53H95NO8/c1-6-9-12-15-18-21-24-27-30-33-36-40-49(55)59-45-53(48-62-52(58)43-39-44-54(4)5,46-60-50(56)41-37-34-31-28-25-22-19-16-13-10-7-2)47-61-51(57)42-38-35-32-29-26-23-20-17-14-11-8-3/h15-20H,6-14,21-48H2,1-5H3/b18-15-,19-16-,20-17-
|
| Chemical Name |
[2-[4-(dimethylamino)butanoyloxymethyl]-3-[(Z)-tetradec-9-enoyl]oxy-2-[[(Z)-tetradec-9-enoyl]oxymethyl]propyl] (Z)-tetradec-9-enoate
|
| HS Tariff Code |
2934.99.9001
|
| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
|
| Solubility (In Vitro) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
|
|---|---|
| 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 | 1.1437 mL | 5.7187 mL | 11.4375 mL | |
| 5 mM | 0.2287 mL | 1.1437 mL | 2.2875 mL | |
| 10 mM | 0.1144 mL | 0.5719 mL | 1.1437 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
