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                            The BI-3802 was designed by Boehringer Ingelheim and could be obtained free of charge through the Boehringer Ingelheim open innovation portal opnMe.com, associated with its negative control.
DODAP is a cationic lipid that is used as a carrier for packaging siRNA and delivering immunostimulatory chemotherapeutic drugs in vivo and in vitro.
Synonyms: 18:1 DAP; 1,2-Dioleoyl-3-dimethylammonium-propane
 
                
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| CAS No. : | 127512-29-2 | 
| Formula : | C41H77NO4 | 
| M.W : | 648.05 | 
| SMILES Code : | O=C(OCC(OC(CCCCCCC/C=C\CCCCCCCC)=O)CN(C)C)CCCCCCC/C=C\CCCCCCCC | 
| Synonyms : | 
                                18:1 DAP; 1,2-Dioleoyl-3-dimethylammonium-propane
                             | 
| MDL No. : | MFCD01321074 | 
| GHS Pictogram: |   | 
| Signal Word: | Warning | 
| Hazard Statements: | H302-H315-H319-H335 | 
| Precautionary Statements: | P261-P280-P301+P312-P302+P352-P305+P351+P338 | 
* All experimental methods are cited from the reference, please refer to the original source for details. We do not guarantee the accuracy of the content in the reference.

| Yield | Reaction Conditions | Operation in experiment | 
|---|---|---|
| 88.9% | 2.41 kg of Nu,Nu'-carbonyldiimidazole is dissolved at room temperature in 6.33 kg of dry acetonitrile. The resultant solution is heated to 25°C. Then 4.0 kg of oleic acid is pumped into the solution over a period of 60 minutes while the reaction temperature is regulated below 35°C by the variation of the addition speed (formation of carbon dioxide gas). After the addition is completed the reaction solution is stirred for additional 90 minutes at 30°C (gas evolution ended). Then 11 g of 1 ,8-diazabicyclo[5.4.0]undec-7-ene is added followed by a solution of 0.83 kg of racemic (R,S)-3-(dimethylamino)- 1 ,2-propanediol in 0.37 kg of dry acetonitrile. Stirring at 30°C is continued for 21 hours. The resultant emulsion is cooled to 25°C and stirring is stopped. Two layers appear. The lower layer is isolated, degassed at 1 mbar/25°C for 200 minutes and finally diluted with 11.7 kg n-heptane. To the solution is added 1.21 kg of basic aluminium oxide and the suspension is stirred for 3 hours at 0°C. The suspension is filtered and the filter residue is washed with 1.5 kg of n-heptane previously cooled down to 0°C. The combined filtrates are homogenized to yield 15.9 kg solution of 4.08 kg pure (2R,S)-DODAP in n-heptane (lot no. MBA-116, assay: 25.7percent, yield:88.9percent). | |
| 11.12 g | With dmap; 1-ethyl-(3-(3-dimethylamino)propyl)-carbodiimide hydrochloride; In chloroform; at 20 - 30℃; for 1h; | <strong>[623-57-4]3-(dimethylamino)-1,2-propanediol</strong> 2.00 g (16.78 mmol, manufactured by Tokyo Chemical Industry Co., Ltd.), Oleic acid 9.48 g (33.56 mmol, Nisshin Oil Co., Ltd. "EXTRA OLEIN 99"), 4-dimethylaminopyridine 0.41 g (3.36 mmol, product of Koei Chemical Industry Co., Ltd.) was dissolved in 120 g of chloroform. There, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride 7.72 g (40.27 mmol, product of Tokyo Chemical Industry Co., Ltd.) And the mixture was stirred at 20 to 30 ° C. After 1 hour, it was washed with 120 g of ion exchange water and 120 g of 25 wtpercent saline, and 3.0 g of anhydrous magnesium sulfate was added to the organic layer and stirred. The magnesium sulfate was filtered off, The filtrate was desolvated with an evaporator to obtain DODAP (yield: 11.12 g, 17.16 mmol). | 
| Yield | Reaction Conditions | Operation in experiment | 
|---|---|---|
| 95% | With N-ethyl-N,N-diisopropylamine; In tetrahydrofuran; at -5 - 20℃;Inert atmosphere; Large scale; | 3-dimethylamino-1,2-propanediol 500 g,1600 g of diisopropylethylamine and 15 L of tetrahydrofuran are added to the reaction vessel.Cool down to minus 5°CThe gas in the replacement axe is nitrogen,A solution of 2800 g of oleic acid chloride in tetrahydrofuran was slowly added dropwise.Keep the temperature in the reactor not higher than 10°C,Then slowly warm up to room temperature,Continue to react for 2-3 hours,Concentrate under reduced pressure,Add Ethyl Acetate and Cold Water,Liquid separation,The organic phase is washed with saturated saline solution.Liquid separation,The organic phase is dried over anhydrous magnesium sulfate,filter.The organic phase is concentrated and dried under reduced pressure to obtain1,2-Dioleoyl-3-diamino-propane2580 g, yield 95percent. | 
| 72% | With pyridine;dmap; In dichloromethane; at 0 - 20℃; for 6h; | An alternative synthesis is as follows: to a solution of 3-(dimethylamino)-l,2- propandiol (0.2 mL, 1.68 mmol), pyridine (0.55 mL, 6.72 mmol) and DMAP (20 mg, 0.17 mmol), in CH2CI2 (10 mL) at 00C was added dropwise oleoyl chloride (1.38 mL, 4.2 mmol). The reaction mixture was allowed to warm up slowly to room temperature. After stirring for 6 hours at room temperature, the reaction mixture was diluted with CH2Cl2 (50 mL), washed sequentially with 5percent aqueous sodium bicarbonate (15 mL), water (15 mL), and saturated aqueous NaCl (15 mL). The organic layer was dried (Na2SO4), filtered and concentrated in vacuo. Purification by column chromatography (SiO2, elution with 0.2percent MeOH in CHCl3) furnished compound II (0.782 g, 72percent) as oily liquid. NMR spectra coordinates are as follows: 1H NMR (CDCl3): delta 5.47-5.32 (m, 4H), 5.28-5.18 (m, IH), 4.40 (dd, J= 11.8, 3.0 Hz, IH)5 4.13 (dd, J= 12.0, 6.8 Hz, IH)5 2.56-2.42 (m, 2H)5 2.35 (dt, J= 7.6, 2.8 Hz, 4H), 2.30 (s, 6H), 2.12-1.97 (m, 8H), 1.71- 1.58 (m, 4H), 1.44-1.23 (m, 40H), 0.92 (t, J= 7.0 Hz, 6H).Another crop of of (+/-)-N,N-Dimethyl-N-[2,3-bis(9-(2)-octadecanoyloxy)- propyl] amine (II) was prepared and had the following 1H NMR characteristics: (400 | 
| With pyridine; In chloroform; at 4℃;Product distribution / selectivity; | In the herein reproduced experiment of Feigner et al., all experimental conditions have been chosen to stay in close accordance with US5,264,618 (Feigner et al.), Example 5, column 27, lines 15 to 47. 31.5 g of oleoylchloride (FLUKA 0733IAH) dissolved in 125 ml chloroform was added dropwise at 4°C under cooling over a period of 11/2 hour to 5.0 g of 3-(dimethylamino)-1 ,2-propanediol, dissolved in 37.5 ml chloroform and 25 ml of pyridine. The yellow solution was stirred overnight. Then 125 ml of cold water and 125 ml diethylether was added. The organic phase was washed twice with 100 ml of 0.5N HCL and also twice with 100 ml 0.5N sodium bicarbonate solution. 39 g anhydrous sodium sulfate was added and the so obtained suspension was filtrated and washed with 100 ml chloroform. The filtrate was then concentrated under reduced pressure at 40°C. 40.1 g of a brown liquid (SM-0318-A) having a (2R,S)-DODAP content of 24.3percent w/w measured by HPLC resulted. A further drying under reduced pressure at 60°C resulted in a reduction of weight to 31.2 g.31.0 g of this material was purified by silicic acid column chromatography as follows:Silica gel: 129 g (the amount of silica gel was calculated relative to the amount (2R,S)-DODAP) Merck 60 F 63-200umColumn: diameter 4 cm, height 60 cmFlow: about 8 ml/minAs mobile phase first 1 ,500 ml methylene chloride (fractions 1-27), then 1 ,000 ml methylene chloride/methanol 95:5 (fractions 28-47) and finally 1 ,000 ml methanol was used. Fractions were collected and combined according to their TLC analysis. So fractions 4-33 were concentrated together under reduced pressure. 10.8 g of a brown oil (SM-0318-B) having a (2R,S)-DODAP content of 65.6percent w/w measured by HPLC resulted. And fractions 34-42 resulted in 12.6 g of a brown oil (SM 0318-D) having a (2R,S)-DODAP content of 54.2percent w/w measured by HPLC.10.4 g Methylene chloride was added to 9.6 g of the compound obtained out of the fractions 4-33 (SM-0318-B) in a high pressure glass tube. The glass tube was then closed and the brownish solution was heated over night at 50°C to form an emulsion. Then the tube was opened and residual methylene chloride was removed by evaporation. 8.0 g of a yellow wax (SM-0318-E) having a (2R,S)-DOTAP chloride content of 65.0percent w/w and 1.3percent w/w (2R,S)-DODAP both measured by HPLC resulted.14.0 g acetonitrile was added to this wax (SM-0318-E). The so obtained emulsion was transferred with 80 ml acetonitrile (to obtain a ratio solid to solvent of about 1 :12) into a flask and cooled down to 20°C. Nocrystallisation could be observed. At 20°C a solidified honey like yellow- brownish material resulted which even when only slightly warming it up tended to become a sticky viscous brownish material.ConclusionThe data demonstrate that 1 ,2-dioleoyl-3-propyltrimethylammonium chloride ((2R,S)-DOTAP chloride) prepared according to the above procedure, which is in accordance with US 5,264,618 (Feigner et al), Example 5, column 27, lines 15 to 47 cannot be obtained in a crystalline form.See also Figures 6 and 7 illustrating (2R,S)-DOTAP chloride emulsion (SM- 0318-E) in acetonitrile when cooling down and (2R,S)-DOTAP chloride emulsion (S -0318-E) in acetonitrile after having been cooled it down to - 20°C, respectively. | 
Tags: DODAP | Liposome | cationic | lipid | ionizable | liposome | encapsulate | siRNA | immunostimulatory chemotherapeutic | inhibitor | 127512-29-2 |
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