ALC-0315 [2036272-55-4]
Referência HY-138170-50mg
Tamanho : 50mg
Marca : MedChemExpress
ALC-0315 is an ionisable aminolipid that is responsible for mRNA compaction and aids mRNA cellular delivery and its cytoplasmic release through suspected endosomal destabilization. ALC-0315 can be used to form lipid nanoparticle (LNP) delivery vehicles. Lipid-Nanoparticles have been used in the research of mRNA COVID-19 vaccine.
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ALC-0315 Chemical Structure
CAS No. : 2036272-55-4
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Based on 7 publication(s) in Google Scholar
Description |
ALC-0315 is an ionisable aminolipid that is responsible for mRNA compaction and aids mRNA cellular delivery and its cytoplasmic release through suspected endosomal destabilization. ALC-0315 can be used to form lipid nanoparticle (LNP) delivery vehicles. Lipid-Nanoparticles have been used in the research of mRNA COVID-19 vaccine[1]. |
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In Vitro |
Preparation of Lipid Nanoparticles Here we provide lipid molar ratios for LNPs in FDA-approved BNT162b2 (a COVID-19 mRNA vaccine). The molar ratio of lipids in this formulation is ALC-0315 : DSPC : Cholesterol : ALC-0159 = 46.3 : 9.4 : 42.7 : 1.6, and RNA to lipid weight ratio is 0.05 (wt/wt) [1] . A. Lipid Mixture Preparation 1. Dissolve lipids in ethanol and prepare 10 mg/m stock solutions. The lipid stock solutions can be stored at −20°C for later use. Note 1: The ionizable lipid is usually a liquid. Due to the viscosity, it should always be weighed rather than relying on the autopipette volume. Note 2: Cholesterol in solution should be kept warm (>37℃) to maintain fluidity. Transfer the cholesterol solution promptly to avoid cooling. 2. Prepare the lipid mixture solution as described. For each mL of lipid mixture add the following: 560 µL of 10mg/mL ALC-0315 (HY-138170), 261 µL of 10mg/mL Cholesterol (HY-N0322), 117 µL of 10mg/mL DSPC (HY-W040193), and 62 µL of ALC-0159 (HY-138300). Mix the solutions thoroµghly to achieve a clear solution. This mixture contains 10 mg of total lipid. Note 3: The choice of lipids and ratios may be changed as desired and this will affect the LNP properties (size, polydispersity, and efficacy) and the amount of mRNA required. B. mRNA Preparation 1. Prepare a 166.7 µg/mL mRNA solution with 100 mM pH 5 sodium acetate buffer. Note 4: The lipid:mRNA weight ratio influences the encapsulation efficiency. Other weight ratios may be prepared as alternative formulations and should be adjusted accordingly by user. C. Mixing There are three commonly used methods to achieve rapid mixing of the solutions: the pipette mixing method, the vortex mixing method, and the microfluidic mixing method. All these mixing methods can be used for various applications. It is important to note that pipette mixing method and vortex mixing method may yield more heterogeneous LNPs with lower encapsulation efficiencies and is prone to variability. Microfluidic devices enable rapid mixing in a highly controllable, reproducible manner that achieves homogeneous LNPs and high encapsulation efficiency. Within these devices, the ethanolic lipid mixture and aqueous solution are rapidly combined in individual streams. LNPs are formed as the two streams mix and are then collected into a single collection tube. 1. Pipette Mixing Method: 1.1. Pipette 3 mL of the mRNA solution and quickly add it into 1 mL of the lipid mixture solution (A 1:3 ratio of ethanolic lipid mixture to aqueous buffer is generally used.) Pipette up and down rapidly for 20–30 seconds. 1.2. Incubate the resulting solution at room temperature for up to 15 minutes. 1.3. After mixing, the LNPs were dialyzed against PBS (pH 7.4) for 2 h, sterile filtered using 0.2 μm filters, and stored at 4°C. 2. Vortex Mixing Method: 1.1. Vortex 3 mL of mRNA solution at a moderate speed on the vortex mixer. Then, Quickly add 1 mL of the lipid mixture solution into the vortexing solution (A 1:3 ratio of ethanolic lipid mixture to aqueous buffer is generally used.). Continue vortexing the resulting dispersion for another 20–30 seconds. 1.2. Incubate the resulting solution at room temperature for up to 15 minutes. 1.3. After mixing, the LNPs were dialyzed against PBS (pH 7.4) for 2 h, sterile filtered using 0.2 μm filters, and stored at 4°C. 3. Microfluidic Mixing Method: 1.1 The 3 mL of mRNA buffer solution and 1 mL of the lipid mixture solution were mixed at a total flow rate of 12 mL/min in a microfluidic device (A 1:3 ratio of ethanolic lipid mixture to aqueous buffer is generally used.). Note 5: Parameters such as the flow rate ratio and total flow rate can be altered to fine-tune LNPs. 1.2. After mixing, the LNPs were dialyzed against PBS (pH 7.4) for 2 h, sterile filtered using 0.2 μm filters, and stored at 4°C. Reference 1. Curr Issues Mol Biol. 2022 Oct 19;44(10):5013-5027. 2. Curr Protoc. 2023;3(9):e898. MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. |
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Masse moléculaire |
766.27 |
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Formule |
C48H95NO5 |
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CAS No. |
2036272-55-4 |
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Appearance |
Liquid |
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Color |
Colorless to light yellow |
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SMILES |
OCCCCN(CCCCCCOC(C(CCCCCC)CCCCCCCC)=O)CCCCCCOC(C(CCCCCC)CCCCCCCC)=O |
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Livraison | Room temperature in continental US; may vary elsewhere. |
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Stockage |
4°C, protect from light *In solvent : -80°C, 6 months; -20°C, 1 month (protect from light) |
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Solvant et solubilité |
In Vitro:
Ethanol : 100 mg/mL (130.50 mM; Need ultrasonic) DMSO : 50 mg/mL (65.25 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO) Preparing
Stock Solutions
View the Complete Stock Solution Preparation Table
*
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol) Concentration (start) × Volume (start) = Concentration (final) × Volume (final) This equation is commonly abbreviated as: C1V1 = C2V2 In Vivo:
Select the appropriate dissolution method based on your experimental animal and administration route.
For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:
Dosage mg/kgAnimal weight Dosing volume Number of animals Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
%
DMSO
+
%
+
%
Tween-80
+
%
Saline
The co-solvents required include: DMSO,
. All of co-solvents are available by MedChemExpress (MCE).
, Tween 80. All of co-solvents are available by MedChemExpress (MCE).
Calculation results:
Working solution concentration:
mg/mL
Method for preparing stock solution:
mg
drug dissolved in
μL
DMSO (Stock solution concentration: mg/mL).
*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)
The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only. If necessary, please contact MedChemExpress (MCE).
Method for preparing in vivo working solution for animal experiments: Take
μL DMSO stock solution, add
μL .
μL , mix evenly, next add
μL Tween 80, mix evenly, then add
μL Saline.
If the continuous dosing period exceeds half a month, please choose this protocol carefully.
Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
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Pureté et documentation | |||||||||||||
Références |
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Complete Stock Solution Preparation Table
*
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Optional Solvent | Concentration Solvent Mass | 1 mg | 5 mg | 10 mg | 25 mg |
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DMSO / Ethanol | 1 mM | 1.3050 mL | 6.5251 mL | 13.0502 mL | 32.6256 mL |
5 mM | 0.2610 mL | 1.3050 mL | 2.6100 mL | 6.5251 mL | |
10 mM | 0.1305 mL | 0.6525 mL | 1.3050 mL | 3.2626 mL | |
15 mM | 0.0870 mL | 0.4350 mL | 0.8700 mL | 2.1750 mL | |
20 mM | 0.0653 mL | 0.3263 mL | 0.6525 mL | 1.6313 mL | |
25 mM | 0.0522 mL | 0.2610 mL | 0.5220 mL | 1.3050 mL | |
30 mM | 0.0435 mL | 0.2175 mL | 0.4350 mL | 1.0875 mL | |
40 mM | 0.0326 mL | 0.1631 mL | 0.3263 mL | 0.8156 mL | |
50 mM | 0.0261 mL | 0.1305 mL | 0.2610 mL | 0.6525 mL | |
60 mM | 0.0218 mL | 0.1088 mL | 0.2175 mL | 0.5438 mL | |
Ethanol | 80 mM | 0.0163 mL | 0.0816 mL | 0.1631 mL | 0.4078 mL |
100 mM | 0.0131 mL | 0.0653 mL | 0.1305 mL | 0.3263 mL |