DOTMA does not have a specific pharmacological target in the traditional sense. It is a cationic lipid that functions as a non-viral gene delivery vehicle. DOTMA carries a positive charge on the surface of liposomes, which condenses anionic nucleic acids (such as DNA, siRNA, and oligonucleotides) and promotes efficient interactions between the cell membrane and the liposome. This facilitates cellular uptake of nucleic acids.

In vitro, DOTMA is an effective transfection agent. It has been used as a component in liposomes that can be used to encapsulate siRNA, microRNAs, and oligonucleotides for gene transfection. DOTMA forms cationic liposomes that condense nucleic acids and promote efficient cellular uptake. These properties make DOTMA a valuable tool for non-viral gene delivery research.

In vivo, DOTMA is an effective transfection agent. It acts as a non-viral vector that enhances the cellular uptake of nucleic acids. DOTMA-based liposomes have been used for in vivo gene delivery applications. However, as a transfection reagent, DOTMA is not a therapeutic agent itself but rather a delivery vehicle for nucleic acid therapeutics.

In vitro enzyme/receptor binding assays are not applicable for DOTMA, as it is a cationic lipid transfection reagent rather than a compound with specific enzyme or receptor targets. Its activity is assessed in cell-based transfection assays rather than biochemical binding assays.

The in vitro cell-based assay for DOTMA involves using it as a transfection reagent to deliver nucleic acids into cultured cells. Cells are seeded in multi-well plates and incubated with liposomes containing DOTMA and the nucleic acid of interest (e.g., plasmid DNA, siRNA, mRNA). Transfection efficiency is assessed by measuring reporter gene expression, target gene knockdown, or protein expression. Cell viability and cytotoxicity are assessed using MTT, LDH release, or CellTiter-Glo assays.

In vivo animal studies for DOTMA involve its use as a component of liposomal formulations for nucleic acid delivery. Animals are administered DOTMA-based liposomes containing therapeutic nucleic acids via various routes (intravenous, intramuscular, intratumoral). Biodistribution, pharmacokinetics, and therapeutic efficacy of the delivered nucleic acids are assessed. Toxicity of the lipid formulation is evaluated through clinical signs, body weight, clinical pathology, and histopathology. Standard protocols for liposome evaluation are employed.

The pharmacokinetic properties of DOTMA as a free lipid are not typically characterized, as it is used as a formulation excipient rather than a therapeutic agent. When administered as part of a liposome, the lipid components are cleared by the liver and spleen and metabolized by phospholipases. The fatty acid components (oleic acid) are utilized for energy or incorporated into cellular membranes. No specific PK data are available.

The toxicity profile of DOTMA has been evaluated as part of liposomal formulations. Cationic lipids can cause cytotoxicity at high concentrations due to membrane disruption. DOTMA-based liposomes may cause immune responses, complement activation, or hepatotoxicity depending on the formulation. Standard toxicology assessments include acute and sub-chronic toxicity studies in rodents. The compound is for research use only and is not intended for human therapeutic use.

1: Gendron F, Grasser M, Le Guennic B. Near-infrared circular dichroism of the ytterbium DOTMA complex: an ab initio investigation. Phys Chem Chem Phys. 2022 Mar 2;24(9):5404-5410. doi: 10.1039/d1cp01675j. PMID: 35170600. 2: Woods M, Payne KM, Valente EJ, Kucera BE, Young VG Jr. Crystal Structures of DOTMA Chelates from Ce3+ to Yb3+ : Evidence for a Continuum of Metal Ion Hydration States. Chemistry. 2019 Jul 25;25(42):9997-10005. doi: 10.1002/chem.201902068. Epub 2019 Jul 3. PMID: 31121070; PMCID: PMC6700027. 3: Mashal M, Attia N, Puras G, Martínez-Navarrete G, Fernández E, Pedraz JL. Retinal gene delivery enhancement by lycopene incorporation into cationic niosomes based on DOTMA and polysorbate 60. J Control Release. 2017 May 28;254:55-64. doi: 10.1016/j.jconrel.2017.03.386. Epub 2017 Mar 24. PMID: 28347807. 4: Kumas C, Fernando WS, Zhao P, Regueiro-Figueroa M, Kiefer GE, Martins AF, Platas-Iglesias C, Sherry AD. Unexpected Changes in the Population of Coordination Isomers for the Lanthanide Ion Complexes of DOTMA-Tetraglycinate. Inorg Chem. 2016 Sep 19;55(18):9297-305. doi: 10.1021/acs.inorgchem.6b01390. Epub 2016 Sep 7. PMID: 27603690; PMCID: PMC5221692. 5: Webber BC, Woods M. Structural analysis of isomeric europium(III) chelates of NB-DOTMA. Inorg Chem. 2012 Aug 6;51(15):8576-82. doi: 10.1021/ic3011597. Epub 2012 Jul 18. PMID: 22809081. 6: Aime S, Botta M, Garda Z, Kucera BE, Tircso G, Young VG, Woods M. Properties, solution state behavior, and crystal structures of chelates of DOTMA. Inorg Chem. 2011 Sep 5;50(17):7955-65. doi: 10.1021/ic2012827. Epub 2011 Aug 5. PMID: 21819052; PMCID: PMC3204394. 7: Matsumoto M, Kishikawa R, Kurosaki T, Nakagawa H, Ichikawa N, Hamamoto T, To H, Kitahara T, Sasaki H. Hybrid vector including polyethylenimine and cationic lipid, DOTMA, for gene delivery. Int J Pharm. 2008 Nov 3;363(1-2):58-65. doi: 10.1016/j.ijpharm.2008.07.010. Epub 2008 Jul 18. PMID: 18687391. 8: Gaucheron J, Santaella C, Vierling P. Transfection with fluorinated lipoplexes based on fluorinated analogues of DOTMA, DMRIE and DPPES. Biochim Biophys Acta. 2002 Aug 31;1564(2):349-58. doi: 10.1016/s0005-2736(02)00469-8. PMID: 12175917. 9: Ren T, Song YK, Zhang G, Liu D. Structural basis of DOTMA for its high intravenous transfection activity in mouse. Gene Ther. 2000 May;7(9):764-8. doi: 10.1038/sj.gt.3301153. PMID: 10822303. 10: Wollenberg B, Kastenbauer, Mundl H, Schaumberg J, Mayer A, Andratschke M, Lang S, Pauli C, Zeidler R, Ihrler S, Löhrs, Naujoks K, Rollston R. Gene therapy --phase I trial for primary untreated head and neck squamous cell cancer (HNSCC) UICC stage II-IV with a single intratumoral injection of hIL-2 plasmids formulated in DOTMA/Chol. Hum Gene Ther. 1999 Jan 1;10(1):141-7. doi: 10.1089/10430349950019273. PMID: 10022539. 11: Konopka K, Davis BR, Düzgüneş N. HIV-1 infection of a non-CD4-expressing variant of HUT-78 cells: lack of inhibition by Leu3A antibodies and enhancement by cationic DOTMA liposomes. Adv Exp Med Biol. 1991;300:97-110. doi: 10.1007/978-1-4684-5976-0_7. PMID: 1781348.

Cationic lipids; structure as described in the first source.

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DOTMA is a research-grade cationic lipid and has not been approved for clinical use as a therapeutic agent. It is one of the first cationic lipids used for gene transfection and is an effective transfection agent in vitro and in vivo. DOTMA carries a positive charge on the surface of liposomes, which condenses anionic nucleic acids and promotes efficient interactions between the cell membrane and the liposome. It is used as a component in liposomes for siRNA, microRNA, and oligonucleotide delivery.

C42H84CLNO2

670.589

669.619

C, 75.23; H, 12.63; Cl, 5.29; N, 2.09; O, 4.77

104162-48-3

104162-48-3 (Z-isomer chloride);122342-03-4 (Z-isomer cation);104872-42-6 (E-isomer chloride);717090-94-3 (E-isomer cation);

6438350

White to off-white solid powder

10.173

0

3

37

46

608

0

[Cl-].CCCCCCCC/C=C\CCCCCCCCOCC(C[N+](C)(C)C)OCCCCCCCC/C=C\CCCCCCCC

LDGWQMRUWMSZIU-LQDDAWAPSA-M

InChI=1S/C42H84NO2.ClH/c1-6-8-10-12-14-16-18-20-22-24-26-28-30-32-34-36-38-44-41-42(40-43(3,4)5)45-39-37-35-33-31-29-27-25-23-21-19-17-15-13-11-9-7-2;/h20-23,42H,6-19,24-41H2,1-5H3;1H/q+1;/p-1/b22-20-,23-21-;

N,N,N-trimethyl-2,3-bis(((Z)-octadec-9-en-1-yl)oxy)propan-1-aminium chloride

DOTMA; 1,2-di-O-octadecenyl-3-trimethylammonium propane

2934.99.9001

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, avoid exposure to moisture.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

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

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.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*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).

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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)]
*Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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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).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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 (Please use freshly prepared in vivo formulations for optimal results.)