Surfactant
20230193137 · 2023-06-22
Inventors
Cpc classification
C09K23/42
CHEMISTRY; METALLURGY
International classification
Abstract
The present invention provides a surfactant of formula (I): (A).sub.m-X-(B).sub.n (I) wherein X is a linking group; each A is N independently a fluorocarbon or a perfluoropolyether; each B is independently (II) wherein a is an integer between 3 and 50 and each R is independently C.sub.1-6 alkyl, CH.sub.2CH.sub.2OC.sub.1-6 alkyl or CH.sub.2CH(CH.sub.3)OC.sub.1-6 alkyl; m is an integer between 1 and 10; and n is an integer between 1 and 10.
##STR00001##
Claims
1. A surfactant of formula (I):
(A).sub.m-X-(B).sub.n (1) wherein X is a linking group; each A is independently a fluorocarbon or a perfluoropolyether; each B is independently ##STR00084## wherein a is an integer between 3 and 50 and each R is independently C.sub.1-6 alkyl, CH.sub.2CH.sub.2OC.sub.1-6 alkyl or CH.sub.2CH(CH.sub.3)OC.sub.1-6 alkyl; m is an integer between 1 and 10; and n is an integer between 1 and 10.
2. A surfactant as claimed in claim 1, which is a star surfactant, preferably a star polymeric surfactant.
3. A surfactant as claimed in claim 1, wherein the sum of n and m is greater than 2, preferably wherein the sum of n and m is an integer between 3 and 10.
4. (canceled)
5. A surfactant as claimed in claim 1, wherein the sum of n and m is 3, 4, 5, 6 or 7, preferably the sum of n and m is 3 or 4, and even more preferably m is 1 and n is 2 or 3, or m is 2 and n is 1 or 2.
6. A surfactant as claimed in claim 1, wherein a is an integer between 3 and 24, preferably 4 and 20.
7. A surfactant as claimed in claim 1, wherein each R is C.sub.1-6 alkyl, preferably methyl.
8. A surfactant as claimed in claim 1, wherein n is an integer between 1 and 6.
9. A surfactant as claimed in claim 1, wherein m is 1, 2 or 3, preferably 1 or 2.
10. (canceled)
11. A surfactant as claimed in claim 1, wherein at least one A, and preferably each A, is a perfluoropolyether.
12. A surfactant as claimed in claim 1, wherein said perfluoropolyether comprises a repeat unit of the formula:
—[CF(CF.sub.3)CF.sub.2O].sub.b—, wherein b is a positive integer, preferably from 1 to 100.
13. A surfactant as claimed in claim 1, wherein said linking group comprises a group selected from —C(O)NH—, —C(O)NMe-, —NHC(O)—, —NMeC(O)—, —C(O)S—, —SC(O)—, —C(O)O—, —OC(O)—, —OC(O)O—, —OC(O)NH—, —OC(O)NMe-, —O—, —S—, —NHC(O)NH—, —NMeC(O)NH—, —NHC(O)NMe-, —NHC(O)O—, —NMeC(O)O—, —SO.sub.2NH—, —NHSO.sub.2—, —NHSO.sub.2—C.sub.6H.sub.4—O— and —O—C.sub.6H.sub.4—SO.sub.2NH—, preferably a group selected from —C(O)NH—, —C(O)NMe-, —NHC(O)—, —NMeC(O)—, —OC(O)NH—, —C(O)O—, —NHC(O)O—, —NMeC(O)O— and —O—.
14. A surfactant as claimed in claim 1, wherein said linking group comprises a charged group, preferably a positively charged group.
15. A surfactant as claimed in claim 1, wherein said linking group comprises a group selected from: ##STR00085## wherein R.sup.1 and R.sup.2 are independently selected from H and C.sub.1-6 alkyl, preferably methyl; and W.sup.− is a counter ion.
16. A surfactant as claimed in claim 1, wherein said linking group comprises a heterocycle and/or a C.sub.1-6 alkylene group, or consists of a C.sub.1-6 alkylene group.
17. (canceled)
18. A surfactant as claimed in claim 1, wherein said linking group comprises a group selected from: ##STR00086## ##STR00087## wherein each p is independently 0 or an integer between 1 and 6; each q is independently 0 or an integer between 1 and 6; each r is independently an integer between 1 and 6; s is an integer between 1 and 6, preferably 1 or 2, still more preferably 1; each Z is independently selected from C(O)NH, C(O)NMe, NHC(O), NMeC(O), OC(O)NH, C(O)O, —NHC(O)O—, —NMeC(O)O— and O; each U is independently a heterocycle; and each Y is independently selected from ##STR00088## wherein W, R.sup.1 and R.sup.2 are as defined in claim 15.
19. A surfactant as claimed in claim 1, wherein said linking group comprises a group of formula (IIIa), (IIIb), (IIIc) or (IIId), preferably a group of formula (IIIb), (IIIc), or (IIId): ##STR00089## wherein p is 0 or 1; q is 0, 1 or 2, preferably 0 or 1; and Z is selected from C(O)NH, C(O)NMe, NHC(O), NMeC(O), OC(O)NH, C(O)O, —NHC(O)O—, —NMeC(O)O— and O; or said linking group comprises a group of formula (IIIe), (IIIf) or (IIIg), preferably (IIIf) or (IIIg): ##STR00090## wherein p is 0 or 1; q is 0, 1 or 2, preferably 0 or 1; r is an integer between 1 and 6, preferably 1, 2 or 3; Z is selected from C(O)NH, C(O)NMe, NHC(O), NMeC(O), OC(O)NH, C(O)O, —NHC(O)O—, —NMeC(O)O— and O; Y is selected from ##STR00091## wherein W, R.sup.1 and R.sup.2 are as defined in claim 15; or said linking group comprises a group of formula (IIIh), (IIIi) or (IIIj), preferably (IIIi) or (IIIj): ##STR00092## wherein p is 0 or 1; q is 0, 1 or 2, preferably 0 or 1; r is an integer between 1 and 6, preferably 1, 2 or 3; Z is selected from C(O)NH, C(O)NMe, NHC(O), NMeC(O), OC(O)NH, C(O)O, —NHC(O)O—, —NMeC(O)O— and O; and U is a heterocycle; or a said linking group comprises a group of formula (IIIk): ##STR00093## wherein each p is 0 or 1; each q is 0, 1 or 2, preferably 0 or 1; and each Z is independently selected from C(O)NH, C(O)NMe, NHC(O), NMeC(O), OC(O)NH, C(O)O, —NHC(O)O—, —NMeC(O)O— and O; or said linking group comprises a group of formula (IIIl) or (IIIm): ##STR00094## wherein each p is 0 or 1; each q is 0, 1 or 2, preferably 0 or 1; each r is an integer between 1 and 6, preferably 1, 2 or 3; each Z is independently selected from C(O)NH, C(O)NMe, NHC(O), NMeC(O), OC(O)NH, C(O)O, —NHC(O)O—, —NMeC(O)O— and O; each U is a heterocycle; and each Y is independently selected from: ##STR00095## wherein W, R.sup.1 and R.sup.2 are as defined in claim 15; or said linking group comprises a group of formula (IIIn): ##STR00096## wherein each p is 0 or 1; each q is 0, 1 or 2, preferably 0 or 1; each r is an integer between 1 and 6; s is an integer between 1 and 6; preferably 1, 2 or 3; each Z is independently selected from C(O)NH, C(O)NMe, NHC(O), NMeC(O), OC(O)NH, C(O)O, —NHC(O)O—, —NMeC(O)O— and O; and each Y is independently selected from: ##STR00097## wherein W, R.sup.1 and R.sup.2 are as defined in claim 15; or said linking group comprises a group of formula (IIIo-q): ##STR00098## wherein each p is independently 0 or an integer between 1 and 6; each q is independently 0 or an integer between 1 and 6; and each Z is independently selected from C(O)NH, C(O)NMe, NHC(O), NMeC(O), OC(O)NH, C(O)O, —NHC(O)O—, —NMeC(O))— and O.
20. (canceled)
21. A surfactant as claimed in claim 1, wherein the ratio of the number of atoms in the backbone structure of the linking group, excluding hydrogen and halogen atoms, as well as any atoms on side or branch groups, to the total number of atoms in the backbone structure of the groups A and B, excluding hydrogen and halogen atoms, as well as any atoms on side or branch groups, is 1:2 to 1:500, more preferably 1:3 to 1:100 and still more preferably 1:5 to 1:50.
22. A surfactant as claimed in claim 1 selected from the group consisting of: ##STR00099## ##STR00100## ##STR00101## ##STR00102## ##STR00103## ##STR00104## ##STR00105##
22. A method for making a surfactant as claimed in claim 1 comprising: reacting a compound of formula (II)
(A).sub.m-X-(OH).sub.n (II) wherein X is a linking group; each A is independently a fluorocarbon or a perfluoropolyether; m is an integer between 1 and 10; n is an integer between 1 and 10; and with a compound of formula (III) ##STR00106## wherein each R is independently C.sub.1-6 alkyl, CH.sub.2CH.sub.2OC.sub.1-6 alkyl or CH.sub.2CH(CH.sub.3)OC.sub.1-6 alkyl; or reacting a compound comprising (A).sub.m with a compound comprising (B).sub.n, wherein said reaction is a coupling reaction and forms linker X between said (A).sub.m and said (B).sub.n.
23. (canceled)
24. A composition, preferably an emulsion, comprising a surfactant of formula (I) as claimed in claim 1.
25-27. (canceled)
28. A method comprising performing one or more chemical and/or biological reactions, and/or biological processes in the discontinuous aqueous phase of an emulsion as claimed in claim 24.
29-39. (canceled)
Description
[0167] The invention will now be described by way of the following non-limiting examples and Figures wherein:
[0168]
[0169]
[0170]
[0171]
[0172]
[0173]
[0174]
EXAMPLES
Example 1: Synthesis of Surfactant 3 Wherein R is Me by Polymerisation
[0175] ##STR00045##
To synthesize surfactant 3, Krytox alcohol (3a) firstly reacts with allyl chloride. The double bond of the resultant intermediate 3b is then oxidized into 1,2-diol, followed by polymer chain extension from the two hydroxyl groups of the Krytox diol intermediate 3c.
Example 2: Synthesis of Surfactant 3 Wherein R is Me by Ether Coupling
[0176] ##STR00046##
Surfactant 3 can also be synthesized starting from polymer chain extension from the two hydroxy of 3-chloro-1,2-propanediol, followed by coupling reaction between the resulting intermediate 3d and Krytox alcohol in the presence of a base.
Example 3: Synthesis of Surfactant 4 Wherein R is Me by Amide Coupling
[0177] ##STR00047##
To synthesize surfactant, 4, 3-chloro-1,2-propanediol is firstly converted into 3-azido-1,2-propanediol followed by polymer chain extension from the two hydroxyl groups. The azido group of the resulting intermediate 4a is reduced into an amino group. Lastly, the intermediate 4b then couples with Krytox acid chloride giving surfactant 4.
Example 4: Synthesis of Surfactant 11 Wherein R is Me by Amide Coupling
[0178] ##STR00048##
The synthesis of surfactant 11 starts from Boc protection of the amino group of Trizma® base, followed by polymer chain extension from the three hydroxyl groups. After the deprotection of Boc of the intermediate 11a, a coupling reaction between the intermediate 11b and Krytox acid chloride gives the target surfactant 11.
Example 5: Synthesis of Surfactant 5 Wherein R is Me by Carbamate Coupling
[0179] ##STR00049##
Surfactant 5 is synthesized by coupling the intermediate 4b with 4-nitrophenyl chloroformate activated Krytox alcohol.
Example 6: Synthesis of Surfactant 8 Wherein R is Me by Coupling
[0180] ##STR00050##
Surfactant 8 is synthesized by coupling the intermediate 3d and the intermediate 8a, which is made by reacting Krytox acid chloride with 1-(2-aminoethyl) piperidine.
Example 7: Synthesis of Surfactant 23 Wherein R is Me by Coupling
[0181] ##STR00051##
The synthesis of surfactant 23 starts from polymer chain extension from the two hydroxyl groups of 2,2-Bis(bromomethyl)-1,3-propanediol giving the intermediate 23a, followed by coupling with the intermediate 23b which is made following the procedure below.
Synthesis of N-[3-(Dimethylamino)propyl] Krytox amide (23b)
[0182] To a stirred solution of 3-(dimethylamino)-1-propylamine (50.61 g, 62.3 mL, 495 mmol) in anhydrous THF (60 mL), ), at 50° C. under nitrogen, was added solution of krytox acid chloride (2, 192.39 g, 82.55 mmol) in Novec 7500 (203 mL) dropwise over 1 H, via canula. After stirring the mixture at 50° C. under nitrogen for 48 hours the mixture was cooled to RT and a yellow solid removed by filtration and washed with Novec 7500 (30 ml). The filtrate was stirred with methanol (4×100 mL), each time the bottom fluorous phase was separated in a separating funnel. The fluorous layer was then evaporated to dryness in vacuo to yield (6) as a pale yellow oil (189.7 g, 95.9%). IR (cm.sup.−1): 2955.5 (bw), 2832.0 (bw), 1729.6 (sm). .sup.1H NMR (400 MHz, 5% C.sub.6D.sub.12 in FC72; vol:vol): 9.506 (1H, bs, NH), 3.493 (2H, m, CONHCH.sub.2), 2.488 (2H, t, CH.sub.2—NMe.sub.2), 2.246 (6H, s, NMe.sub.2), 1.692 (2H, m, CH.sub.2—CH.sub.2NMe.sub.2).
Example 8: Synthesis of Surfactant 27 Wherein R is Me by Coupling
[0183] ##STR00052##
The synthesis of surfactant 27 starts from polymer chain extension from the two hydroxyl groups of 1,4-Dibromo-2,3-butanediol giving the intermediate 27a, followed by coupling with the intermediate 23b which is made following the procedure above.
Example 9: Synthesis of Surfactant 34 Wherein R is Me by Click Chemistry
[0184] ##STR00053##
Firstly, Krytox amide was reduced to Krytox amine, followed by coupling with 4-nitrophenyl chloroformate activated (1R)-(−)-nopol. The resulting intermediate 34a then undergoes Click reaction with intermediate 4a giving the target surfactant 34.
Example 10: Synthesis of Surfactant 36 Wherein R is Me by Click Chemistry
[0185] ##STR00054##
Firstly, 2,3-dibromo-1-propanol reacts with sodium azide giving 2,3-diazido-1-propanol, followed by polymer chain extension. The resulting intermediate 36a then undergoes Click reaction with Krytox nopol (36b), which is made from Krytox alcohol, giving the target surfactant 36.
Example 11: Synthesis of Surfactant 35 Wherein R is Me by Click Chemistry
[0186] ##STR00055##
The synthesis of surfactant 35 starts from the reaction between 4-nitrophenyl chloroformate activated (1R)-(−)-nopol and the intermediate 11b. The resulting intermediate 35a then undergoes Click reaction with Krytox azide (35b), which is made from Krytox alcohol, giving the target surfactant 35.
Example 12
Synthesis of Krytox-FSL-PEG4-11-azido-1-amide
[0187] ##STR00056##
To a stirred solution of Krytox FSL 2,2,2-trifluoroethyl ester (57.26 g, 23.97 mmol) in Novec 7100 (58.0 mL), under nitrogen at 38° C., was added a solution of 11-azido-3,6,9-trioxaundecan-1-amine (5.393 g, 24.71 mmol) in anhydrous THF (25.5 mL) via syringe, including a 1.5 mL wash. Then stirred overnight at 40° C., where TLC indicated that a lot unreacted amine was present. Triethylamine (3.512 mL, 25.2 mmol) was added via syringe and the block temperature raised to 60° C. After 48 hours the reaction was cooled to RT and evaporated to dryness in vacuo. The resulting oil was dissolved in Novec 7500 (200 mL) and was extracted in a separating funnel with methanol (4×50 mL). The fluorous layer was then evaporated in vacuo to give a thick yellow oil (69.0 g). IR (cm.sup.−1) 2115.0 (N.sub.3), 1790.5 (C═O, TFE-ester), 1730.62 (C═O, product). The two carbonyl stretches were roughly equal in intensity. A portion of the crude material (20.95 g) was dissolved in Novec 7100 (15.0 mL) and loaded on to an 80 g Interchim 50 μm HC cartridge, pre-washed with Novec 7100. The column was then washed as follows: a. Novec 7100 (250 mL), b. 0.5% MeOH/Novec 7100 (250 mL), c. 1.0% MeOH/Novec 7100 (250 mL), d. 1.5% MeOH/Novec 7100 (250 mL), e. 2.0% MeOH/Novec 7100 (250 mL), f. 3.0% MeOH/Novec 7100 (250 mL), g. 5.0% MeOH/Novec 7100 (250 mL), h. 10.0% MeOH/Novec 7100 (500 mL). The starting material eluted in Novec 7100 and was evaporated in vacuuo to give a colourless oil (12.224 g). The late running product band fractions were combined and evaporated in vacuo to give the product as a clear oil (7.317 g, 12.15%). IR (cm.sup.−1): 2112.3 (N.sub.3, mBr), (C═O, 1717.15). NMR (400 MHz, 5% C.sub.6D.sub.12 in FC-72 (weight:weight) containing 0.04% TMS as internal standard, Δ (ppm) 7.80 (1H, s, NH), 3.72 (14H, s), 3.60 (2H, s), 3.35 (2H, s).
Polymerisation of 1-(2-(6,6-dimethylbicyclo[3.1.1]hept-2-en-2-yl)ethoxy)-poly(2-methoxymethylethylene oxide)
[0188] ##STR00057##
t-Bu-P4 Phosphazene base (10.48 ml, 0.8 M in Hexane, 8.4 mmol) was added to a solution of (1R)-(−)-Nopol (1.40 g, 8.4 mmol) in toluene (40 ml). Methyl Glycidyl Ether (19.6 g, 222.4 mmol) was added dropwise and the resultant mixture stirred at 40° C. under N2 for 2 days. The reaction was quenched with benzoic acid (2.0 g), concentrated under reduced pressure and re-dissolved in THF. The product was purified by passing though neutral alumina, filtering and concentrating under reduced pressure to give a pale brown oil (11.33 g).
Click Reaction Between Krytox-FSL-PEG4-11-azido-1-amide and 1-(2-(6,6-dimethylbicyclo[3.1.1]hept-2-en-2-yl)ethoxy)-poly(2-methoxymethylethylene oxide)
[0189] ##STR00058##
To a solution of the crude nopol polymer (0.93 g) in Novec 7500 (5.4 mL) was added a solution of Krytox-FSL-PEG.sub.4-11-azido-1-amide (1.303 g, 5.18 mmol) in anhydrous DMF (2.0 mL), via syringe and under a nitrogen atmosphere. The heating block was set to 105° C. After 3 hours TLC (4% MeOH in Novec 7100 indicated that all the azide had reacted, but heating was continued at 105° C. overnight. On cooling to RT, Novec 7500 (20 mL) was added and the solution extracted in a separating flask with methanol (3×50 mL). The fluorous phase was evaporated to dryness in vacuo to give the product (1.156 g). A portion of the crude material (1.00 g) was dissolved in Novec 7100 (3.0 mL) and loaded on to an 25 g Interchim 50 μm HC cartridge, pre-washed with Novec 7100. The column was then washed as follows: a. Novec 7100 (100 mL), b. 1.0% MeOH/Novec 7100 (100 mL), c. 2.0% MeOH/Novec 7100 (100 mL), d. 3.0% MeOH/Novec 7100 (100 mL), e. 4.0% MeOH/Novec 7100 (100 mL), f. 5.0% MeOH/Novec 7100 (100 mL), g. 6.0% MeOH/Novec 7100 (100 mL), h. 7.0% MeOH/Novec 7100 (100 mL) and i. 8.0% MeOH/Novec 7100 (100 mL). The polar fractions were combined and evaporated in vacuo to give the product as a pale-yellow oil (365 mg). A 5.0% stock solution was made up by dissolving the oil in Novec 7500 (6.935 g) and used for microdroplet generation.
Example 13
2,2-Bis(azidomethyl)-1,3-propane-diol
[0190] ##STR00059##
To solid 2,2-Bis(azidomethyl)-1,3-propanediol (24.06 g, 91.86 mmol) was added a solution of sodium azide (18.19 g, 275.6 mmol) in water (50 mL), at RT under nitrogen, the remaining azide beaker was washed out with water (40.0 mL) and added to the reaction. The stirred solution was then heated to a block temperature of 100° C., under nitrogen, as the solution was slightly hazy, extra water (30.0 mL) was added and the block temperature increased to 105° C. After 46 hours the reaction was cooled to RT and extracted with DCM (3×100 mL). The combined organic extracts were dried over sodium sulphate, filtered and evaporated to give the product as a pale-yellow oil (16.24 g, 95.0%). NMR (400 MHz, CDCl.sub.3 containing 0.04% TMS as internal standard, Δ (ppm) 3.65 (4H, t, CH.sub.2—O), 3.43 (4H, t, CH.sub.2—N.sub.3), 2.60 (2H, bs, OH).
2,2-Bis(azidomethyl)-1,3-propane-di-O-polymer
[0191] ##STR00060##
To a stirred solution of 2,2-Bis(azidomethyl)-1,3-propane-diol (200 mg, 1.075 mmol) in anhydrous toluene (10 mL), under nitrogen at RT, was added a solution P.sub.4-t-Bu in hexane (0.8 M, 2.68 mL, 2.15 mmol, phosphazene base) via syringe. Then neat 2-(methoxymethyl)oxirane (2.215 mL, 36.49 mmol, 34.0 mol equivalents) was added to the stirring solution via syringe at RT. After 48 hours the reaction was quenched by the addition of solid benzoic acid (0.934 g, 7.65 mmol), following dissolution the reaction was evaporated in vacuo. The crude product was dissolved in DCM (50 mL) and washed with water (50 mL), the aqueous phase was back extracted with DCM (2×15 mL). The combined organic phase was washed with saturated sodium bicarbonate. The organic fraction was then stirred with aminopropyl silica (1.64 g, 1.78 mmol) for ten minutes, then filtered and evaporated in vacuo to give a pale-yellow oil (2.985 g). The oil was dissolved in THF (3.0 mL) and loaded on to a gravity column of alumina (34 g), pre-washed with THF. The column was washed with first THF (300 mL), which was evaporated in vacuo to give a yellow oil (0.722 g, IR (cm−1) 2101.0 (weak, s, N.sub.3). Washing the column with 10% methanol in THF (250 mL) gave the crude product (1.876 g, IR (cm.sup.−1) 2101.0 (medium, s, N.sub.3).
2,2-Bis(aminomethyl)-1,3-propane-di-O-polymer
[0192] ##STR00061##
To a solution of the crude 2,2-bis(aminomethyl)-1,3-propane-di-O-polymer (0.86 g, ca. 0.27 mmol) in methanol (3.0 mL) was added solid ammonium formate (85 mg, 1.35 mmol), followed by zinc dust (118 mg, 1.80 mmol) and the reaction starred at RT under nitrogen. After 20 hours the solids were filtered off over celite and the filtrate evaporated in vacuo to give a pale-yellow oil (0.731 g). The oil was dissolved on to a 1 g SCX cartridge and eluted first with methanol (15 mL) and then with ammonia in methanol (7.0 M, 15.0 mL). The methanolic ammonia fraction was evaporated to give a green oil (90 mg).
Synthesis of Surfactant 22 Wherein R is Me
[0193] ##STR00062##
To a stirred solution of the 2,2-bis(aminomethyl)-1,3-propane-di-O-polymer (90 mg, 28.8 mmol) in THF (2.5 mL) was added N-methylmorpholine (50 pL, 0.54 mmol) at RT under nitrogen. A solution of Krytox-4-nitrophenyl carbonate (133 mg, 71.94 μmol, 2.5 mol equivalents) in Novec 7100 (3.0 mL) was added, during the addition the solution went from green to yellow. The block temperature then raised to 40° C. and stirred for ca. 112 hours before cooling to RT. The solution was evaporated to dryness in vacuo, then dissolved in Novec 7500 (12.0 mL) and washed with methanol (3×10.0 mL). The fluorous fraction was evaporated in vacuo to give the crude product as a colourless oil (219 mg). IR (cm.sup.−1) 1779.8 (weak, s, unreacted Krytox carbonate) and 1744.6 (medium, s, carbamate).
Example 14: Water Picodroplet Generation
[0194] In order to generate droplets with volumes between 300-400 pL, fluorous oil (e.g. Novec 7500) containing 5% (w/w) of the surfactant synthesised in example 12, was used as the continuous carrier oil phase, whilst a biochemical buffer solution was used as the dispersed aqueous phase. The two phases were infused using Cetoni GmbH syringe pumps connected via polythene tubing (ID: 0.38 mm) to PDMS microfluidic chip having a single flow-focusing nozzle (nozzle dimensions: 60×60 μm). Typical flow rates ranged between 1000-1500 μL/hr for the fluorous phases and were kept constant at 500 μL/hr for aqueous phases.
The Figures show that the picodroplets are stabilised by the surfactant.
Example 15: Synthesis of Surfactant 37 Wherein R is Me
Synthesis of 2,3-diazidopropan-1-ol
[0195] ##STR00063##
2,3-dibromopropan-1-ol (4.36 g, 20 mmol) was added to a stirring solution of NaN.sub.3 in DMSO (120 mL, 60 mmol) and stirred at 75° C. for 2 days. TLC showed only 1 product spot. The reaction was quenched with water (250 mL), extracted with diethyl ether (3×, 150 mL), washed with water (2×, 150 mL) and saturated brine (150 mL), and dried with sodium sulphate (50 g). Filtration yields the product as a yellow oil (2.231 g, 15.7 mmol, 79%). IR (cm.sup.−1) 3374.8 (broad, m, OH), 2932.8 (b, m, CH), 2085.0 (Sharp, s, N.sub.3). NMR (400 MHz, CDCl.sub.3 containing 0.04% TMS as internal standard, Δ (ppm) 3.82 (3H, m, CH.sub.2OH & CHN.sub.3), 3.45 (1H, m, CH.sub.2N.sub.3), 2.00 (1H, s, OH).
Synthesis of 1-(2,3-diazidopropoxy)-poly(3-methoxypropyleneoxide)
[0196] ##STR00064##
2-(methoxymethyl)oxirane (8.38 g, 95.1 mmol) was added to a solution of 2,3-diazido-propan-1-ol (0.51 g, 3.59 mmol) in toluene (20 mL). Tert-Bu-P4 base (4.49 mL, 0.8 M in hexane, 3.59 mmol) was added dropwise and the mixture stirred at room temperature for 6 days. The reaction was quenched with benzoic acid (1.00 g), and evaporated to yield a crude oil (8.183 g). This was purified by column chromatography eluted with THF (100%) giving the product (2.989 g). NMR (400 MHz, CDCl.sub.3 containing 0.04% TMS as internal standard, Δ (ppm) 3.94 (2H, broad s, 2H, CH.sub.2OH), 3.64 (broad m, 4H), 3.56-3.40 (m, 7.5 H), 3.39-3.36 (m, 5H), 1.89 (2H, s, OH).
Synthesis of 1-(2,3-diaminopropoxy)-poly(3-methoxypropyleneoxide)
[0197] ##STR00065##
1-(2,3-diazidopropoxy)-poly(3-methoxypropyleneoxide) (2.0 g) was added to a stirring solution of triphenyl phosphine (1.20 g, 4.56 mmol) in methanol (50 mL) and heated to 50° C. The solution was stirred under N.sub.2 at 50° C. for two weeks. The mixture was evaporated and redissolved in dichloromethane (30 mL). This was acidified with HCl (1.0 M, 15 mL) and brine (5 mL), washed with dichloromethane (2×15 mL), basified with NaOH (10 M) and extracted with dichloromethane (3×20 mL). The combined organic fractions were dried over Na.sub.2SO.sub.4 and evaporated to yield the product as a yellow/brown oil (0.31 g). NMR (400 MHz, CDCl.sub.3 containing 0.04% TMS as internal standard, Δ (ppm) 3.94 (2H, s, CH.sub.2O), 3.68 (26H, broad s), 3.55-3.28 (60H, broad m), 2.80 (4H, bm, (NH.sub.2).sub.2), 0.8 (2H, broad s, OH).
Synthesis of Surfactant 37
[0198] ##STR00066##
[0199] A solution of 1-(2,3-diaminopropoxy)-poly(3-methoxypropyleneoxide) (0.294 g, 0.639 mmol) and 4-methylmorphline (0.21 mL, 1.1917 mmol) in THF (25 mL) was added to a solution of PFPE-CH.sub.2OC(O)O-Ph-4-NO.sub.2 (2.36 g, 1.278 mmol) in Novec 7100 (15 mL) and stirred under N.sub.2 at 40° C. for 1 week. The reaction mixture was filtered, evaporated and redissolved in Novec 7100. The solution was then washed, and subsequently filtered with aminopropyl functionalized silica (20 g, 10 g and 10 g) until no yellow colour remained. The resultant solution was evaporated to yield a pale yellow oil (0.765 g) which was purified on a 20 g silica (30 um) column chromatography eluted with 2% MeOH/Novec 7100 (75 mL), 5% MeOH/Novec 7100 (50 mL), 7% MeOH/Novec 7100 (50 mL) and 10% MeOH/novec 7100 (100 mL) yielding the product (0.667 g). NMR (400 MHz, 5% C.sub.6D.sub.12 in FC-72 (weight:weight) containing 0.04% TMS as internal standard, Δ (ppm) 4.78 (4H, broad m, (Krytox-CH.sub.2).sub.2), 4.10-3.70 (broad m, 16H), 3.68-3.40 (broad m, 9.2 H), 3.39-3.25 (broad m, 16H).
Microdroplet Generation
[0200] In order to generate droplets with volumes between 300-400 pL, fluorous oil (e.g. Novec 7500) containing 5% (w/w) of surfactant 37 synthesised in Example 15, was used as the continuous carrier oil phase, whilst a cell culturing medium was used as the dispersed aqueous phase. The two phases were infused using Cetoni GmbH syringe pumps connected via polythene tubing (ID: 0.38 mm) to PDMS microfluidic chip having a single flow-focusing nozzle (nozzle dimensions: 60×60 μm). Typical flow rates ranged between 800-1000 μL/hr for the fluorous phases and were kept constant at 500 μL/hr for aqueous phases.
Example 16: Synthesis of Surfactant 38 Wherein R is Me
Synthesis of 2,3-diazidobutane-1,4-diol
[0201] ##STR00067##
A solution of 2,3-dibromo-1,4-butanediol (4.96 g, 20 mmol) in water (25 mL) was added to a solution of NaN.sub.3 (3.90 g, 60 mmol) in water (25 mL), and heated under N.sub.2 at 80° C. for 5 days. The reaction mixture was cooled to room temperature, washed with chloroform/isopropanol (3:1, 4×50 mL). The combined organic extracts were dried over Na.sub.2SO.sub.4 and evaporated to yield a yellow oil (2.95 g, 17.2 mmol, 86%).
Synthesis of 1,4-(2,3-diazido butoxy)-dipoly(3-methoxypropyleneoxide)
[0202] ##STR00068##
2-(methoxymethyl)oxirane (13.57 g, 154.0 mmol) was added to a solution of 2,3-diazidobutane-1,4-diol (1.00 g, 5.81 mmol) in toluene (50 mL). Tert-Bu-P4 base (7.26 mL, 5.81 mmol) was added dropwise yielding a change from colourless to clear yellow. The reaction was stirred at room temperature for 1 week under N.sub.2. The reaction was quenched with benzoic acid (1.50 g), and evaporated. The crude oil was purified by column chromatography through neutral alumina eluted with THF (100%) giving the product (8.389 g).
Synthesis of 1,4-(2,3-diamino butoxy)-dipoly(3-methoxypropyleneoxide)
[0203] ##STR00069##
1,4-(2,3-diazido butoxy)-dipoly(3-methoxypropyleneoxide) (4.0 g) was added to a stirring solution of triphenyl phosphine (1.20 g, 4.56 mmol) in methanol (100 mL). Water (0.82 g, 45.6 mmol) was added and the solution heated to 50° C., stirred under N.sub.2 at 50° C. for two days. The mixture was evaporated and redissolved in dichloromethane (30 mL). This was acidified with HCl (1.0 M, 3×15 mL) and brine (5 mL), washed with dichloromethane (2×15 mL), basified with NaOH (10 M) and extracted with dichloromethane (3×20 mL). The combined organic fractions were dried over Na.sub.2SO.sub.4 and evaporated to yield the product as a yellow oil (1.538 g).
Synthesis of Surfactant 38
[0204] ##STR00070##
[0205] A solution of 1,4-(2,3-diamino butoxy)-dipoly(3-methoxypropyleneoxide) (0.8 g, 0.735 mmol) and 4-methylmorphline (0.22 g, 2.206 mmol) in THF (25 mL) was added to a solution of PFPE-CH.sub.2OC(O)O-Ph-4-NO.sub.2 (2.71 g, 1.471 mmol) in Novec 7100 (15 mL) and stirred under N.sub.2 at room temperature for 1 week. The reaction mixture was evaporated and redissolved in Novec 7100 (50 mL), followed by washing with 2-aminopropyl functionalized silica (2×20 g) until no yellow colour remained. The resultant solution was evaporated to yield a pale yellow oil (0.43 g) which was purified on a 20 g silica (30 um) column chromatography eluted with 2% MeOH/Novec 7100 (75 mL), 5% MeOH/Novec 7100 (50 mL), 7% MeOH/Novec 7100 (50 mL) and 10% MeOH/novec 7100 (100 mL) yielding the product.
Example 17: Synthesis of Surfactant 39 Wherein R is Me
Polymerisation of 1-(2-(6,6-dimethylbicyclo[3.1.1]hept-2-en-2-yl)ethoxy)-poly(2-methoxymethylethylene oxide)
[0206] ##STR00071##
t-Bu-P4 Phosphazene base (10.48 ml, 0.8 M in Hexane, 8.4 mmol) was added to a solution of (1R)-(−)-Nopol (1.40 g, 8.4 mmol) in toluene (40 ml). Methyl Glycidyl Ether (19.6 g, 222.4 mmol) was added dropwise and the resultant mixture stirred at 40° C. under N2 for 2 days. The reaction was quenched with benzoic acid (2.0 g), concentrated under reduced pressure and redissolved in THF. The product was purified by passing though neutral alumina, filtering and concentrating under reduced pressure to give a pale brown oil (11.33 g).
Alternative Method
[0207] To a solution of (1R)-(−)-Nopol (0.181 g, 1.09 mmol) in toluene (2.6 ml) was added 2-(methoxymethyl)glycidol (0.959 g, 10.89 mmol) followed by a solution of aluminium triflate (0.0181 g, 3.817×10.sup.−5 mol) in dioxane (0.31 mL) and the solution was warmed on a heating block at 75° C. After 18 hours the reaction mixture was allowed to cool to room temperature to give a light brown oil (1.011 g) and columned on neutral alumina (12 g) and eluted with THF. Fractions 1-3 were combined and evaporated to dryness to give a pale-yellow oil (0.198 g). NMR (400 MHz, CDCl.sub.3 containing 0.04% TMS as internal standard, Δ (ppm) 5.107 (0.39 H, m, Nopol=C—H, minor isomer), 5.022 (0.61 H, m, Nopol=C—H, major isomer), 4.35-4.22 (m, 1H), 4.12 (m, 1H), 3.956-3.71 (m, 5H), 3.67-3.40 (m, 8H), 3.395-3.383 (m, OMe, 14.6 H, n=4.86), 2.10-1.80 (m, 3.0H), 1.776 (1H, s, OH), 1.73-1.51 (m, 3H), 1.292-1.09 (m, 4H), 1.07-0.95 (m, 4H), 0.85-0.80 (m, 2H).
Synthesis of Krytox-FSL-PEG4-11-azido-1-amide
[0208] ##STR00072##
To a stirred solution of Krytox FSL 2,2,2-trifluoroethyl ester (57.26 g, 23.97 mmol) in Novec 7100 (58.0 mL), under nitrogen at 38° C., was added a solution of 11-azido-3,6,9-trioxaundecan-1-amine (5.393 g, 24.71 mmol) in anhydrous THF (25.5 mL) via syringe, including a 1.5 mL wash. Then stirred overnight at 40° C., where TLC indicated that some of the unreacted amine was still present. Triethylamine (3.512 mL, 25.2 mmol) was added via syringe and the block temperature raised to 60° C. After 48 hours the reaction was cooled to RT and evaporated to dryness in vacuuo. The resulting oil was dissolved in Novec 7500 (200 mL) and was extracted in a separating funnel with methanol (4×50 mL). The fluorous layer was then evaporated in vacuuo to give a thick yellow oil (69.0 g). IR (cm.sup.−1) 2115.0 (N.sub.3), 1790.5 (C═O, TFE-ester), 1730.62 (C═O, product). The two carbonyl stretches were roughly equal in intensity. A portion of the crude material (20.95 g) was dissolved in Novec 7100 (15.0 mL) and loaded on to an 80 g Interchim 50 μm HC cartridge, pre-washed with Novec 7100. The column was then washed as follows: a. Novec 7100 (250 mL), b. 0.5% MeOH/Novec 7100 (250 mL), c. 1.0% MeOH/Novec 7100 (250 mL), d. 1.5% MeOH/Novec 7100 (250 mL), e. 2.0% MeOH/Novec 7100 (250 mL), f. 3.0% MeOH/Novec 7100 (250 mL), g. 5.0% MeOH/Novec 7100 (250 mL), h. 10.0% MeOH/Novec 7100 (500 mL). The starting material eluted in Novec 7100 and was evaporated in vacuuo to give a colourless oil (12.224 g). The late running product band fractions were combined and evaporated in vacuuo to give the product as a clear oil (7.317 g, 12.15%). IR (cm.sup.−1): 2112.3 (N.sub.3, mBr), (C═O, 1717.15). NMR (400 MHz, 5% C.sub.6D.sub.12 in FC-72 (weight:weight) containing 0.04% TMS as internal standard, Δ (ppm) 7.80 (1H, s, NH), 3.72 (14H, s), 3.60 (2H, s), 3.35 (2H, s).
Alternative Procedure
[0209] To a stirred solution of Krytox FSL acid chloride (24.46 g, 12.25 mmol, IR 1806.7 cm.sup.−1) in Novec 7500 (50.0 mL), under nitrogen at room temperature, was added a solution of 11-azido-3,6,9-trioxaundecan-1-amine (2.674 g, 12.25 mmol) in anhydrous 1,4-dioxane (20.0 mL, plus 5.0 mL wash) via syringe. Then stirred overnight at 40° C., where TLC indicated that some of the unreacted amine was still present. To the resulting solution was added PS-piperdine (3.0-4.0 mmol/g, 5.249 g) and the reaction temperature raised to 75° C. and stirred for 72 hours. The reaction was cooled to RT and the polystyrene beads removed by filtration, the reaction flask and beads were washed with Novec 7500 (2×40 mL). The filtrate was shaken with methanol (30 mL) and bottom fluorous layer separated and evaporated in vacuo to mixture of Krytox methyl ester (30.5%) and the desired Krytox-FSL-PEG4-11-azido-1-amide (26.36 g) as judged by proton NMR. %). IR (cm.sup.−1): 2109.9 (N.sub.3), 1785.7 (CO.sub.2Me), 1723.0 (CONH).
Synthesis of Surfactant 39
[0210] Click reaction between Krytox-FSL-PEG4-11-azido-1-amide and 1-(2-(6,6-dimethylbicyclo[3.1.1]hept-2-en-2-yl)ethoxy)-poly(2-methoxymethylethylene oxide)
##STR00073##
To a solution of the crude nopol polymer (0.93 g) in Novec 7500 (5.4 mL) was added a solution of Krytox-FSL-PEG.sub.4-11-azido-1-amide (1.303 g, 5.18 mmol) in anhydrous DMF (2.0 mL), via syringe and under a nitrogen atmosphere. The heating block was set to 105° C. After 3 hours TLC (4% MeOH in Novec 7100 indicated that all the azide had reacted, but heating was continued at 105° C. overnight. On cooling to RT, Novec 7500 (20 mL) was added and the solution extracted in a separating flask with methanol (3×50 mL). The fluorous phase was evaporated to dryness in vacuuo to give the product (1.156 g). A portion of the crude material (1.00 g) was dissolved in Novec 7100 (3.0 mL) and loaded on to a 25 g Interchim 50 μm HC cartridge, pre-washed with Novec 7100. The column was then washed as follows: a. Novec 7100 (100 mL), b. 1.0% MeOH/Novec 7100 (100 mL), c. 2.0% MeOH/Novec 7100 (100 mL), d. 3.0% MeOH/Novec 7100 (100 mL), e. 4.0% MeOH/Novec 7100 (100 mL), f. 5.0% MeOH/Novec 7100 (100 mL), g. 6.0% MeOH/Novec 7100 (100 mL), h. 7.0% MeOH/Novec 7100 (100 mL) and i. 8.0% MeOH/Novec 7100 (100 mL). The polar fractions were combined and evaporated in vacuuo to give the product as a pale-yellow oil (365 mg). A 5.0% stock solution was made up by dissolving the oil in Novec 7500 (6.935 g) and used for microdroplet generation.
Example 18: Synthesis of Surfactant 40 Wherein R is Me
2,2-Bis(azidomethyl)-1,3-propane-diol
[0211] ##STR00074##
To solid 2,2-Bis(azidomethyl)-1,3-propanediol (24.06 g, 91.86 mmol) was added a solution of sodium azide (18.19 g, 275.6 mmol) in water (50 mL), at RT under nitrogen, the remaining azide beaker was washed out with water (40.0 mL) and added to the reaction. The stirred solution was then heated to a block temperature of 100° C., under nitrogen, as the solution was slightly hazy, extra water (30.0 mL) was added and the block temperature increased to 105° C. After 46 hours the reaction was cooled to RT and extracted with DCM (3×100 mL). The combined organic extracts were dried over sodium sulphate, filtered and evaporated to give the product as a pale-yellow oil (16.24 g, 95.0%). NMR (400 MHz, CDCl.sub.3 containing 0.04% TMS as internal standard, Δ (ppm) 3.65 (4H, s, CH.sub.2—O), 3.43 (4H, s, CH.sub.2—N.sub.3), 2.60 (2H, bs, OH).
2,2-Bis(azidomethyl)-1,3-propane-di-O-poly(2-methoxymethylethylene oxide)
[0212] ##STR00075##
To a stirred solution of 2,2-Bis(azidomethyl)-1,3-propane-diol (200 mg, 1.075 mmol) in anhydrous toluene (10 mL), under nitrogen at RT, was added a solution P.sub.4-t-Bu in hexane (0.8 M, 2.68 mL, 2.15 mmol, phosphazene base) via syringe. Then neat 2-(methoxymethyl)oxirane (2.215 mL, 36.49 mmol, 34.0 mol equivalents) was added to the stirring solution via syringe at RT. After 48 hours the reaction was quenched by the addition of solid benzoic acid (0.934 g, 7.65 mmol), following dissolution the reaction was evaporated in vacuo. The crude product was dissolved in DCM (50 mL) and washed with water (50 mL), the aqueous phase was back extracted with DCM (2×15 mL). The combined organic phase was washed with saturated sodium bicarbonate. The organic fraction was then stirred with aminopropyl silica (1.64 g, 1.78 mmol) for ten minutes, then filtered and evaporated in vacuuo to give a pale-yellow oil (2.985 g). The oil was dissolved in THF (3.0 mL) and loaded on to a gravity column of alumina (34 g), pre-washed with THF. The column was washed with first THF (300 mL), which was evaporated in vacuuo to give a yellow oil (0.722 g, IR (cm−1) 2101.0 (weak, s, N.sub.3). Washing the column with 10% methanol in THF (250 mL) gave the crude product (1.876 g, IR (cm−1) 2101.0 (medium, s, N.sub.3).
2,2-Bis(aminomethyl)-1,3-propane-di-O-poly(2-methoxymethylethylene oxide)
[0213] ##STR00076##
To a solution of the crude 2,2-bis(azidoomethyl)-1,3-propan-di-O-poly(2-methoxymethylethylene oxide) (2.564 g, ca. 1.74 mmol) in methanol (35.0 mL) was added triphenylphosphine (0.913 g, 3.48 mmol). Then water (0.626 mL) was added and the heating block was warmed to 77° C. and left to stir for 19 hours. The solution was evaporated to dryness to give an oily solid (3.325 g) that was dissolved in DCM and washed with 2.0 M HCl (1×10 mL) and (1×20 mL). The combined acid layer was neutralised with sodium hydroxide (10 M, 10.62 mL) and was extracted with DCM (3×20 mL). The combined organic fractions were dried over sodium sulphate filtered and evaporated to give a pale-yellow oil (1.132 g). NMR (400 MHz, CDCl.sub.3 containing 0.04% TMS as internal standard, Δ (ppm) 3.95 (broad singlet), 3.8-3.40 (broad multiplet), 3.378 (—OMe, broad singlet).
Alternative Method
[0214] To a stirred solution of the 2,2-bis(azidoomethyl)-1,3-propan-di-O-poly(2-methoxymethylethylene oxide) (0.86 g, ca. 0.27 mmol)(columned on neutral alumina) in methanol (3.0 mL), under nitrogen at room temperature, was added first ammonium formate (solid, 85.2 mg, 1.35 mmol) and then zinc dust (<10 μm, 110 mg, 1.68 mmol). The crude reaction mixture was filtered over celite and washed with methanol and the filtrate evaporated in vacuo to give a pale-yellow oil (0.731 g). The crude oil was purified on a 1 g SCX cartridge, eluting first with methanol and then ammonia in methanol (7 M, 15 mL), which was concentrated in vacuo to give a green oil (90 mg).
Surfactant 40—2,2-Bis(krytox carboxamido-methyl)-1,3-propan-di-O-poly(2-methoxymethylethylene oxide)
[0215] ##STR00077##
To a stirred solution of the crude 2,2-bis(aminomethyl)-1,3-propane-di-O-poly(2-methoxymethylethylene oxide) (0.705 g) in dioxane (anhydrous, 17.5 mL), under nitrogen, was added PS-piperidine (0.652 g, ca. 3.5 mmol/g). A solution of Krytox acid chloride (3.416 g) in Novec7500 (15.0 mL, plus 2.5 mL wash) was then added via syringe. The aluminium block temperature was raised to 80° C. and allowed to stir for approximately 60 hours. The reaction mixture was then allowed to cool to RT, filtered and the filtrate transferred to a separating funnel. The lower fluorous layer was separated from the dioxane layer and extracted once with methanol (25 mL). The fluorous layer was separated and evaporated to dryness to give a pale yellow oil (3.456 g). IR (cm−1) 1790.7 (weak, s, CO, ester) and 1738.9 (weak, sharp, CO, amide). The crude material was columned on a silica cartridge (25 g) and eluted with 1% methanol in Novec7100 to 15% methanol in Novec7100 stepwise. The late running fractions were evaporated to give a brown oil (0.54 g), IR (cm−1) 3369.8 (weak, b, —OH), 2891.3 (strong, br, CH), 1699.6 (strong, s, CO, amide). NMR (400 MHz, CDCl.sub.3 containing 0.04% TMS as internal standard, Δ (ppm) 5.35 (broad multiplet, NH), 3.82 (s, CH.sub.2O), 3.77 (s, CH.sub.2O), 3.72 (s, CH.sub.2O), 3.72 (s, CH.sub.2O), 3.7-3.5 (broad multiplet, CHO), 3.46 (s, OMe), 3.45-3.10 (broad multiplet, CHO). Sample contains P.sub.4-t-Bu impurity.
Example 19 Alternative Synthesis of Surfactant 22 Wherein R is Me
[0216] ##STR00078##
To a stirred solution of 4-nitrophenyl chloroformate (25.24 g, 125.22 mmol) in THF (54.5 mL), at room temperature under nitrogen, was added Novec7100 (32.0 mL) via syringe. A solution of Krytox alcohol (52.63 g, 31.3 mmol) was then added via syringe in a mixture of Novec7100 (15.0 mL) and THF (24.5 mL). Finally a solution of pyridine (5.1 mL, 62.6 mmol) in a mixture of Novec7100 (4.0 mL) and THF (6.5 mL) was added via syringe, where upon a white solid immediately started to precipitate out of solution. The reaction was then warmed to 40° C. and allowed to stir for 48 hours. The reaction was cooled to room temperature and filtered over celite and the bed washed with Novec7100 (50 mL). The filtrate was evaporated in vacuo and dissolved in Novec7100:FC72 (1:1, vol:vol, 200 mL), where upon a precipitate started to form. After 1 hour the solution was filtered over celite and the bed washed with a little FC72 (30 mL). The filtrate was concentrated in vacuo and redissolved in FC72 (180 mL) and allowed to stand overnight. Finally the solution was filtered over celite and evaporated to dryness to give a colourless oil (54.88 g, 95.0%). NMR (400 MHz, 5% C.sub.6D.sub.12 in FC-72 (weight:weight) containing 0.04% TMS as internal standard, Δ (ppm) 8.21 (2H, d, aromatic), 7.30 (2H, d, aromatic), 4.80 (2H, m, CH.sub.2).
##STR00079##
To a stirred solution of the crude 2,2-bis(aminomethyl)-1,3-propane-di-O-polymer (1.072 g, ca. 0.87 mmol) in THF (anhydrous, 12.5 mL), under nitrogen at room temperature, was added N-methylmorpholine (0.48 mL, 4.35 mmol) via syringe. Then a solution of Krytox 4-nitrophenyl carbonate (3.53 g, 19.15 mmol) was added in Novec 7100 (14.0 mL, plus 1.0 ml wash) was added via syringe and the heating block set to 42° C. and allowed to stir for 48 hours. The reaction was cooled to room temperature and was evaporated to dryness in vacuo to give an oil, which was washed with THF (2×20 mL) and decanted off the residue. The residue was dissolved in Novec7500 (50 mL) and extracted with methanol (1×50 mL and 2×30 mL). The fluorous phase was concentrated in vacuo to give a dark oil (4.54 g). The oil was dissolved in Novec7100 (50 mL), with stirring and aminopropyl silica (2.52 g, 1.78 mml/g) was added and allowed to stir for 30 minutes. The mixture was filtered and the silica washed with a little Novec7100 (10 mL). The filtrate was treated with aminopropyl silica (2.52 g, 1.78 mml/g) two more times and after 30 minutes the solution was filtered and the silica washed with a little Novec7100 (10 mL). After the third filtration the filtrate was concentrated under low vacuum. The resulting oil was dissolved in Novec7100 (4.0 mL) and loaded on to a silica cartridge (25 g, SIHC, Interchim) packed in Novec7100. A gradient from Novec7100 to 15% methanol in 7100 was run collecting 50 mL fractions. Fractions 12 and 13 were combined and concentrated in vacuo to yield a light-brown oil, fraction A (0.84 g). IR (cm.sup.−1) 1737.9 (Krytox-CH.sub.2C═O, carbamate). NMR (400 MHz, 5% C.sub.6D.sub.12 in FC-72 (weight:weight) containing 0.04% TMS as internal standard, Δ (ppm) 5.47 (4H, s, Krytox-CH.sub.2), 5.05 (6H, s), 4.78 (12H, m), 4.00-3.48 (68H, m), 3.47-3.25 (47.5H, bs, OMe). Fractions 14-18 were concentrated in vacuo to give a light brown oil, fraction B (0.685 g). IR (cm.sup.−1) 1768.3 (Krytox-CH.sub.2O, carbonate). NMR 400 MHz, 5% C.sub.6D.sub.12 in FC-72 (weight:weight) containing 0.04% TMS as internal standard, Δ (ppm) 5.45 (4H, s, Krytox-CH.sub.2O), 5.05 (10H, s) 4.75 (25H, s), 4.00-3.48 (96H), 3.47-3.28 (100H, bs, OMe).
Example 20
[0217] ##STR00080##
To a stirred solution of Krytox acid chloride (12.893 g, 5.54 mmol) in Novec7100 was added a solution of dipropargylamide (0.60 mL, 5.81 mmol) in anhydrous THF (4.0 mL, plus 1.00 mL wash), under nitrogen at room temperature, where a fine white precipitate started to form. Finally, a solution of triethylamine (1.16 mL, 8.30 mmol) in anhydrous THF (4.0 mL) was added and the solution warmed to 55° C. After 20 hours the reaction was cooled to RT and filtered over celite and washed with Novec7100 (20 mL) and the filtrate evaporated to dryness in vacuo to yield a light brown oil (13.009 g, 98.5%). IR (cm−1) 3320.2 (weak, CH), 1697.3 (medium, s, CO). NMR (400 MHz, CDCl.sub.3 containing 0.04% TMS as internal standard, Δ (ppm) (4H, dt, N(CH2)2), 2.08 (2H, d, CH).
1-Azido-3-methoxy-propan-2-ol & 2-azido-3-methoxy-propan-1-ol
[0218] ##STR00081##
To a stirred solution of tetra-n-butylammonium azide (5.245 g, 18.44 mmol) in 1,2-DME (74.0 mL), at room temperature under nitrogen, was added a solution of 2-(methoxymethyl)oxirane (1.547 g, 17.56 mmol) in 1,2-DME (anhydrous, 9.0 mL plus 1.0 mL wash) via syringe. Finally, a solution of aluminium triflate (10 mM, 1.756 mmol) in 1,2-DME was added and the heating block temperature was raised to 65° C. The reaction was stirred for 48H and then allowed to cool to RT. The crude reaction mixture was cooled to room temperature and concentrated in vacuo to give a yellow oil (7.60 g). Multiple chromatography using ethyl acetate in hexane gradients finally gave the desired mixture of diastereomers free of tetra-n-butylammonium salts as a pale-yellow oil (0.798 g, 34.9%). IR (cm.sup.−1) 3421.3 (OH, bm), 2927.7 (CH, bm), 2094.0 (N.sub.3, ss). NMR (400 MHz, CDCl.sub.3 containing 0.04% TMS as internal standard, Δ (ppm) 3.94 (1H, m, CH—OH), 3.55-3.42 (2H, m, CH.sub.2OMe), 3.40 (3H, s, OMe), 3.38-3.32 (2H, m, CH.sub.2—N3), 2.46 (1H, s, OH).
1-Azido-3-methoxy-propan-2-O-poly(2-methoxymethylethylene oxide)
[0219] ##STR00082##
To a stirred solution of tetra-n-butylammonium azide (1.493 mmol, 0.25 M) in 1,2-DME (5.97 mL), at room temperature under nitrogen, was added a solution of 2-(methoxymethyl)oxirane (1.973 g, 22.40 mmol) in 1,2-DME (anhydrous, 4.0 mL plus 1.0 mL wash) via syringe. Finally, a solution of aluminium triflate (10 mM, 44.8 μmol, 4.48 mL) in 1,2-DME was added and the heating block temperature was raised to 75° C. The reaction was stirred for 48H and then allowed to cool to RT. The crude reaction mixture was cooled to room temperature and concentrated in vacuo to give a yellow oil (2.1 g). Purification on a 40 g silica cartridge (SIHC, Interchim) using ethyl acetate in hexane gave the desired azido-polymer (0.473 g, 33.7%, n=10.2, mw 941, using internal standard) as a pale-yellow oil. IR (cm.sup.−1) 3460.0 (OH, bm), 2877.7 (CH, bm), 2097.2 (N.sub.3, ss). NMR (400 MHz, CDCl.sub.3 containing 0.04% TMS as internal standard, Δ (ppm) 4.03-3.93 (1H, m, CH—OH), 3.77-3.54 (6H, m, CH.sub.2OMe), 3.53-3.42 (12.5H, s, OMe), 3.38-3.32 (2H, m, CH.sub.2—N3), 1.96 (1H, s, OH).
Krytox-bis-Click Surfactant 41
[0220] ##STR00083##
To a stirred solution of 1-Azido-3-methoxy-propan-2-O-poly(2-methoxymethylethylene oxide)(0.257 g, Mn 836, 0.307 mmol) in t-butanol (2.70 mL), under nitrogen at 36° C., was added a solution of Krytox dipropargylamide (1.10 g, 0.4611 mmol) in Novec7100 (5.0 mL). Then a solution of copper (II) acetate:THPTA (1:1, 9.357 μmol each) in t-butanol:water (5:1, vol:vol, 0.783 mL) was added via pipette. Next morning the reaction was cooled to room temperature and evaporated to dryness in vacuo to a blue/green oil (1.294 g), which was dissolved in Novec7100 and purified on a 25 g silica cartridge (SiHC, Interchim) and eluted with a gradient of 100% Novec7100 To 12% MeOH in Novec7100. The later running fractions were combined to give the bis-1,4-triazole surfactant (0.884 g). NMR 400 MHz, 5% C.sub.6D.sub.12 in FC-72 (weight:weight) containing 0.04% TMS as internal standard, Δ (ppm) 8.08 (2H, broad m, (triazole-H5).sub.2), 5.40-4.26 (5H, broad m), 4.25-3.90 (3H, broad m), 3.90-3.15 (25H, broad m, OMe), 1.37 (1H, broad m, OH), 0.85 (1H, broad m, OH).