Biodegradable polyionenes

Abstract

Synthesis of polyionenes with built-in degradable linkers through addition polymerization of a novel class of degradable A.sub.2-type monomers (d-A.sub.2), and their use as antimicrobial agents are disclosed. A library of biodegradable polyionenes and Gemini-surfactants made from d-A.sub.2 monomers are also disclosed. These materials have potent and broad spectrum of antimicrobial activity with high selectivity over mammalian cells.

Claims

1. A compound having the following formula (I), (Ia) or (Ib): ##STR00040## ##STR00041## wherein A is independently selected from the group consisting of optionally substituted alkoxy, optionally substituted aminoalkoxy, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted cycloalkynyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkenyl, optionally substituted heterocycloalkynyl, optionally substituted aryl and optionally substituted heteroaryl; R.sup.1 is independently selected from the group consisting of optionally substituted C.sub.2-C.sub.15 alkyl, optionally substituted C.sub.2-C.sub.15 alkenyl, optionally substituted C.sub.2-C.sub.15 alkynyl, —Ar—, —R.sup.2—Ar—R.sup.2—, —R.sup.2—(O—R.sup.2).sub.m—R.sup.2—, —R.sup.2—N(R.sup.4)—R.sup.2— and —R.sup.2—C(R.sup.4).sub.2—R.sup.2—; Ar is independently optionally substituted aryl or optionally substituted heteroaryl; R.sup.2 is independently optionally substituted alkyl, optionally substituted alkenyl or optionally substituted alkynyl; X.sup.1 is independently halogen, sulfate, tosylate, mesylate, ##STR00042## X.sup.2 is independently absent or an anionic counterion; R.sup.3 is independently selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted alkenyl and optionally substituted alkynyl; R.sup.4 is independently selected from the group consisting of hydrogen, R.sup.3, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted cycloalkynyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkenyl, optionally substituted heterocycloalkynyl, optionally substituted amino, optionally substituted alkylsulfide, —R.sup.2—OC(O)—R.sup.3, —R.sup.2—C(O)O—R.sup.3, —R.sup.2—Ar and —R.sup.2—O—R.sup.2—CH.sub.2═CH.sub.2; R.sup.5 is independently —R.sup.2— or —R.sup.2—O—R.sup.2—; Z is independently O or S; Q is independently O or NH; n is an integer from 1 to 3; m and q are independently an integer from 1 to 10; p′ is 0 or is an integer of at least 1; p is 0 or in an integer of at least 1; and r is 0 or 1, wherein when p is 0, then r is 1; or when p is an integer of at least 1, then r is 0 or 1.

2. The compound according to claim 1, wherein Z is O and Q is NH, or Z is O and Q is O, or A is Ar, or n is 1 or m is 3.

3. The compound according to claim 1, wherein R.sup.2 is a C.sub.1 to C.sub.15 linear alkyl or C.sub.1 to C.sub.15 linear or branched alkenyl or wherein R.sup.1 is selected from the group consisting of: —(CH.sub.2).sub.2—, —(CH.sub.2).sub.3—, —(CH.sub.2).sub.4—, —(CH.sub.2).sub.5—, —(CH.sub.2).sub.6—, —(CH.sub.2).sub.7—, —(CH.sub.2).sub.8—, —CH.sub.2—O—CH.sub.2CH.sub.2—O—CH.sub.2—, —(CH.sub.2).sub.2—O—CH.sub.2CH.sub.2—O—(CH.sub.2).sub.2—, —CH.sub.2—{O—CH.sub.2CH.sub.2}2-O—CH.sub.2—, —(CH.sub.2).sub.2—{O—CH.sub.2CH.sub.2}2-O—(CH.sub.2).sub.2—, —(CH.sub.2).sub.3—{O—CH.sub.2CH.sub.2}2—O—(CH.sub.2).sub.3—, ##STR00043## CH.sub.2—C(NH.sub.2)—CH.sub.2—, —CH.sub.2—CH(CH.sub.3)(NH.sub.2)—CH.sub.2—, 9-fluorenylmethyl carbamate (Fmoc), t-butyl carbamate(Boc), benzyl carbamate (Cbz), acetyl (Ac), trifluoroacetyl, benzyl (Bn), trityl (Tr), benzylideneamine, p-toluenesulfonamide (Ts), and p-methoxyphenyl (PMP).

4. The compound according to claim 1, wherein X.sup.1 is halogen or X.sup.1 is ##STR00044## or X.sup.1 is ##STR00045##

5. The compound according to claim 1, wherein R.sup.3 is alkyl or wherein R.sup.4 is selected from the group consisting of H, —(CH.sub.2).sub.2, —(CH.sub.2).sub.3, —(CH.sub.2).sub.4, —(CH.sub.2).sub.5, —(CH.sub.2).sub.6, —(CH.sub.2).sub.7, —(CH.sub.2).sub.8, —(CH.sub.2).sub.9, —(CH.sub.2).sub.10, —(CH.sub.2).sub.11, —(CH.sub.2).sub.12, —(CH.sub.2).sub.13, —(CH.sub.2).sub.14, —(CH.sub.2).sub.15, —CH.sub.2CH═CH, —(CH.sub.2).sub.2—CH═CH, —(CH.sub.2).sub.3—CH═CH, —(CH.sub.2).sub.4—CH═CH, —(CH.sub.2).sub.5—CH═CH, —(CH.sub.2).sub.6—CH═CH, —(CH.sub.2).sub.7—CH═CH, —(CH.sub.2).sub.8—CH═CH, —(CH.sub.2).sub.9—CH═CH, —(CH.sub.2).sub.10—CH═CH, —(CH.sub.2).sub.11—CH═CH, —(CH.sub.2).sub.13—CH═CH, —CH.sub.2—OC(O)—CH.sub.2, —CH.sub.2—OC(O)—(CH.sub.2).sub.2, —CH.sub.2—OC(O)—(CH.sub.2).sub.3, —CH.sub.2—OC(O)—(CH.sub.2).sub.4, —(CH.sub.2).sub.2—OC(O)—CH.sub.2, —(CH.sub.2).sub.2—OC(O)—(CH.sub.2).sub.2, —(CH.sub.2).sub.2—OC(O)—(CH.sub.2).sub.3, —(CH.sub.2).sub.2—OC(O)—(CH.sub.2).sub.4, —(CH.sub.2).sub.2—OC(O)—C(═CH.sub.2)CH.sub.3, —CH.sub.2—C(O)O—CH.sub.2, —CH.sub.2—C(O)O—(CH.sub.2).sub.2, —CH.sub.2—C(O)O—(CH.sub.2).sub.3, —CH.sub.2—C(O)O—(CH.sub.2).sub.4, —(CH.sub.2).sub.2—C(O)O—CH.sub.2, —(CH.sub.2).sub.2—C(O)O—(CH.sub.2).sub.2, —(CH.sub.2).sub.2—C(O)O—(CH.sub.2).sub.3, —(CH.sub.2).sub.2—C(O)O—(CH.sub.2).sub.4, —(CH.sub.2).sub.2—C(O)O—C(═CH.sub.2)CH.sub.3, —NH.sub.2, —SH, ##STR00046## or wherein R.sup.5 is selected from the group consisting of —(CH.sub.2).sub.2—, —(CH.sub.2).sub.3—, —(CH.sub.2).sub.4—, —(CH.sub.2).sub.5—, —(CH.sub.2).sub.6—, —(CH.sub.2).sub.7—, —CH═CH—, —CH═CH—CH.sub.2—, —CH.sub.2—CH═CH—, —CH═CH—(CH.sub.2).sub.2—, —CH.sub.2—CH═CH═CH.sub.2—, —(CH.sub.2).sub.2—CH═CH—, —CH═CH═CH—, —CH═CH—(CH.sub.2).sub.3—, —CH.sub.2—CH═CH—(CH.sub.2).sub.2—, —(CH.sub.2).sub.2—CH═CH—CH.sub.2—, —(CH.sub.2).sub.3—CH═CH—, —CH═CH═CH—CH.sub.2—, —CH.sub.2—CH═CH═CH—, —CH.sub.2—O—CH.sub.2—, —CH.sub.2—O—(CH.sub.2).sub.2—, —(CH.sub.2).sub.2—O—CH.sub.2—, —(CH.sub.2)—O—(CH.sub.2).sub.3—, —(CH.sub.2).sub.2—O—(CH.sub.2).sub.2—, and —(CH.sub.2).sub.3—O—CH.sub.2—.

6. The compound according to claim 1, wherein X.sup.2 is selected from the group consisting of halogen, sulfate, tosylate, and mesylate.

7. The compound according to claim 1, wherein p is 0, having the following formula (III), (IIIa) or (IIIb): ##STR00047## wherein X.sup.3 is halogen, or having the following formula (IV), (IVb), or (IVc): ##STR00048##

8. The compound according to claim 1 wherein p is an integer of at least 1, having the following formula (V), (Va), (Vb), (Vc), (Vd) or (Ve): ##STR00049## wherein X.sup.3 is halogen.

9. The compound according to claim 1, wherein when p and p′ are both an integer of at least 1, the p′:p ratio is in the range of about 1:10 to about 10:1, having a molecular weight in the range of about 1 kDa to about 100 kDa or having a molar mass dispersity in the range of about 1.1 to about 4.0.

10. A method for making a compound according to claim 1, the method comprising the steps of: (A) contacting NH.sub.2—R.sup.1—NH.sub.2 or HO—R.sup.1—OH with a compound having the following formula (VI): ##STR00050## wherein R.sup.1 is selected from the group consisting of optionally substituted C.sub.2-C.sub.15 alkyl, optionally substituted C.sub.2-C.sub.15 alkenyl, optionally substituted C.sub.2-C.sub.15 alkynyl, —Ar—, —R.sup.2—Ar—R.sup.2—, —R.sup.2—(O—R.sup.2).sub.m—R.sup.2—, —R.sup.2—N(R.sup.4)—R.sup.2— and —R.sup.2—C(R.sup.4).sub.2—R.sup.2—; Ar is independently optionally substituted aryl or optionally substituted heteroaryl; R.sup.2 is independently optionally substituted alkyl, optionally substituted alkenyl or optionally substituted alkynyl; Z is independently O or S; A is independently selected from the group consisting of optionally substituted alkoxy, optionally substituted aminoalkoxy, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted cycloalkynyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkenyl, optionally substituted heterocycloalkynyl, optionally substituted aryl and optionally substituted heteroaryl; n is an integer from 1 to 3; m is an integer from 1 to 10; and X.sup.3 is halogen; to form a halogen compound having the following formula (III): ##STR00051## or (B) contacting ##STR00052## with a compound having the following formula (VIa): ##STR00053## wherein R.sup.1 is selected from the group consisting of optionally substituted C.sub.2-C.sub.15 alkyl, optionally substituted C.sub.2-C.sub.15 alkenyl, optionally substituted C.sub.2-C.sub.15 alkynyl, —Ar—, —R.sup.2—Ar—R.sup.2—, —R.sup.2—(O—R.sup.2).sub.m—R.sup.2—, —R.sup.2—N(R.sup.4)—R.sup.2— and —R.sup.2—C(R.sup.4).sub.2—R.sup.2—; Ar is independently optionally substituted aryl or optionally substituted heteroaryl; R.sup.2 is independently optionally substituted alkyl, optionally substituted alkenyl or optionally substituted alkynyl; Z is independently O or S; A is independently selected from the group consisting of optionally substituted alkoxy, optionally substituted aminoalkoxy, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted cycloalkynyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkenyl, optionally substituted heterocycloalkynyl, optionally substituted aryl and optionally substituted heteroaryl; n is an integer from 1 to 3; m is an integer from 1 to 10; and X.sup.3 is halogen; to form a halogen compound having the following formula (IIIa) or (IIIb): ##STR00054## wherein Q is independently O or NH.

11. The method according to claim 10, comprising the step of contacting the halogen compound having the formula (III), (IIIa), or (IIIb) with an amine N(R.sup.3).sub.2R.sup.4: wherein R.sup.3 is independently selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted alkenyl and optionally substituted alkynyl; and R.sup.4 is independently selected from the group consisting of hydrogen, R.sup.3, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted cycloalkynyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkenyl, optionally substituted heterocycloalkynyl, optionally substituted amino, optionally substituted alkylsulfide-R.sup.2—OC(O)—R.sup.3, —R.sup.2—C(O)O—R.sup.3, —R.sup.2—Ar and —R.sup.2—O—R.sup.2—CH.sub.2═CH.sub.2; to form a tertiary amino compound having the following formula (IV), (IVb) or (IVc): ##STR00055## wherein X.sup.2 is absent or an anionic counterion.

12. The method according to claim 10, comprising the step of contacting the halogen compound having the formula (III), (IIIa) or (IIIb) with a diamine (R.sup.3).sub.2 N—R.sup.2—N(R.sup.3).sub.2: wherein R.sup.3 is independently selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted alkenyl and optionally substituted alkynyl; to form a polymer product having the following formula (V), (Va), (Vb), (Vc), (Vd) or (Ve) ##STR00056## wherein X.sup.2 is absent or an anionic counterion; X.sup.3 is halogen; R.sup.5 is —R.sup.2— or —R.sup.2—O—R.sup.2—; q is an integer from 1 to 10; p′ is 0 or an integer of at least 1; and r is 0 or 1, wherein when p is an integer of at least 1, then r is 0 or 1.

13. The method according to claim 12, comprising the steps of: contacting the halogen compound having the formula (III), (IIIa) or (IIIb) with the diamine (R.sup.3).sub.2 N—R.sup.2—N(R.sup.3).sub.2, wherein R.sup.3 is as defined in claim 12; and adding another halogen compound having the formula (III), (IIIa) or (IIIb); to form the polymer product having the following formula (V), (Va), (Vb), (Vc), (Vd) or (Ve) ##STR00057##

14. A composition comprising the compound having the following formula (I), (Ia) or (Ib): ##STR00058## wherein A is independently selected from the group consisting of optionally substituted alkoxy, optionally substituted aminoalkoxy, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted cycloalkynyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkenyl, optionally substituted heterocycloalkynyl, optionally substituted aryl and optionally substituted heteroaryl; R.sup.1 is independently selected from the group consisting of optionally substituted C.sub.2-C.sub.15 alkyl, optionally substituted C.sub.2-C.sub.15 alkenyl, optionally substituted C.sub.2-C.sub.15 alkynyl, —Ar—, —R.sup.2—Ar—R.sup.2—, —R.sup.2—(O—R.sup.2).sub.m—R.sup.2—, —R.sup.2—N(R.sup.4)—R.sup.2— and —R.sup.2—C(R.sup.4).sub.2—R.sup.2—; Ar is independently optionally substituted aryl or optionally substituted heteroaryl; R.sup.2 is independently optionally substituted alkyl, optionally substituted alkenyl or optionally substituted alkynyl; X.sup.1 is independently halogen, sulfate, tosylate and mesylate, ##STR00059## X.sup.2 is independently absent or an anionic counterion; R.sup.3 is independently selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted alkenyl and optionally substituted alkynyl; R.sup.4 is independently selected from the group consisting of hydrogen, R.sup.3, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted cycloalkynyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkenyl, optionally substituted heterocycloalkynyl, optionally substituted amino, optionally substituted alkylsulfide, —R.sup.2—OC(O)—R.sup.3, —R.sup.2—C(O)O—R.sup.3, —R.sup.2—Ar and —R.sup.2—O—R.sup.2—CH.sub.2═CH.sub.2; R.sup.5 is independently —R.sup.2— or —R.sup.2—O—R.sup.2—; Z is independently O or S; Q is independently O or NH; n is an integer from 1 to 3; m and q are independently an integer from 1 to 10; p′ is 0 or is an integer of at least 1; p is 0 or an integer of at least 1; and r is 0 or 1, wherein when p is 0, then r is 1; or when p is an integer of at least 1, then r is 0 or 1.

15. The composition according to claim 14, wherein the compound has the following formula (IV), (IVb) or (IVc): ##STR00060##

16. The composition according to claim 14, wherein the compound has the following formula (V), (Va), (Vb), (Vc), (Vd) or (Ve): ##STR00061## wherein X.sup.3 is halogen.

17. The composition according to claim 14 further comprising a thickening agent and/or a detergent.

18. The composition according to claim 17, wherein the composition comprises the compound in the range of about 0.2 wt % to about 1 wt %, a thickening agent in the range of about 0.5 wt % to about 1.5 wt %, and a detergent in the range of about 3 wt % to about 15 wt %, with the remaining wt % being water so that the total is 100 wt %.

19. A method for treating a bacterial infection or fungal infection comprising administering to a subject a compound according to claim 1.

Description

BRIEF DESCRIPTION OF DRAWINGS

(1) The accompanying drawings illustrate a disclosed embodiment and serves to explain the principles of the disclosed embodiment. It is to be understood, however, that the drawings are designed for purposes of illustration only, and not as a definition of the limits of the invention.

(2) FIG. 1 refers to a schematic diagram showing the synthesis of degradable polyionenes, P and Gemini-surfactants S synthesized through novel A.sub.2-type bis-halide monomers d-A.sub.2 and other readily commercially available reagents such as di-tertiary amines B.sub.2 and functional tertiary amines B, respectively.

(3) FIG. 2 refers to a schematic guide and chemical structures of the different types of d-A.sub.2 monomers synthesized, the chemical structures of other difunctional monomers and monofunctional reagents used for the synthesis of the polyionenes (P) and surfactants (S).

(4) FIG. 3 is a .sup.1H NMR spectrum of monomer d-A.sub.2-1.

(5) FIG. 4 is a .sup.1H NMR spectrum of monomer d-A.sub.2-2

(6) FIG. 5 is a .sup.1H NMR spectrum of monomer d-A.sub.2-3.

(7) FIG. 6 is a .sup.1H NMR spectrum of monomer d-A.sub.2-4.

(8) FIG. 7 is a .sup.1H NMR spectrum of monomer d-A.sub.2-5.

(9) FIG. 8 is a .sup.1H NMR spectrum of monomer d-A.sub.2-6.

(10) FIG. 9 is a .sup.1H NMR spectrum of monomer d-A.sub.2-7.

(11) FIG. 10 is a .sup.1H NMR spectrum of monomer d-A.sub.2-8.

(12) FIG. 11 is a .sup.1H NMR spectrum of monomer d-A.sub.2-9.

(13) FIG. 12 is a .sup.1H NMR spectrum of monomer d-A.sub.2-10.

(14) FIG. 13 is a .sup.1H NMR spectrum of monomer d-A.sub.2-11.

(15) FIG. 14 is a .sup.1H NMR spectrum of monomer d-A.sub.2-12.

(16) FIG. 15 is a .sup.1H NMR spectrum of monomer d-A.sub.2-13.

(17) FIG. 16 is a .sup.1H NMR spectrum of monomer d-A.sub.2-14.

(18) FIG. 17 is a .sup.1H NMR spectrum of monomer d-A.sub.2-15.

(19) FIG. 18 is a .sup.1H NMR spectrum of monomer d-A.sub.2-16.

(20) FIG. 19 is a .sup.1H NMR spectrum of monomer d-A.sub.2-17.

(21) FIG. 20 is a .sup.1H NMR spectrum of monomer d-A.sub.2-18.

(22) FIG. 21 is a .sup.1H NMR spectrum of monomer d-A.sub.2-19.

(23) FIG. 22 is a .sup.1H NMR spectrum of monomer d-A.sub.2-20.

(24) FIG. 23 is a .sup.1H NMR spectrum of P-1a-1.

(25) FIG. 24 is a .sup.1H NMR spectrum of P-1c-1.

(26) FIG. 25 is a .sup.1H NMR spectrum of P-3a.

(27) FIG. 26 is a .sup.1H NMR spectrum of P-3c.

(28) FIG. 27 is a .sup.1H NMR spectrum of P-5c-1.

(29) FIG. 28 is a .sup.1H NMR spectrum of P-6a.

(30) FIG. 29 is a .sup.1H NMR spectrum of P-6b.

(31) FIG. 30 is a .sup.1H NMR spectrum of P-6c.

(32) FIG. 31 is a .sup.1H NMR spectrum of P-4a.

(33) FIG. 32 is a .sup.1H NMR spectrum of P-4b.

(34) FIG. 33 is a .sup.1H NMR spectrum of P-4c.

(35) FIG. 34 is a .sup.1H NMR spectrum of P-1b.

(36) FIG. 35 is a .sup.1H NMR spectrum of P-5a.

(37) FIG. 36 is a .sup.1H NMR spectrum of P-5b.

(38) FIG. 37 is a .sup.1H NMR spectrum of P-21a.

(39) FIG. 38 is a .sup.1H NMR spectrum of P-7a.

(40) FIG. 39 is a .sup.1H NMR spectrum of P-(7:1/1:1)a.

(41) FIG. 40 is a .sup.1H NMR spectrum of P-(7:1/3:7)a.

(42) FIG. 41 is a .sup.1H NMR spectrum of P-(7:1/1:9)a.

(43) FIG. 42 is a .sup.1H NMR spectrum of P-8a.

(44) FIG. 43 is a .sup.1H NMR spectrum of P-8b.

(45) FIG. 44 is a .sup.1H NMR spectrum of P-8c.

(46) FIG. 45 is a .sup.1H NMR spectrum of P-(8:21/1:1)a.

(47) FIG. 46 is a .sup.1H NMR spectrum of P-(8:21/1:9)a.

(48) FIG. 47 is a .sup.1H NMR spectrum of P-9a.

(49) FIG. 48 is a .sup.1H NMR spectrum of P-10a.

(50) FIG. 49 is a .sup.1H NMR spectrum of P-12a.

(51) FIG. 50 is a .sup.1H NMR spectrum of P-13a.

(52) FIG. 51 is a .sup.1H NMR spectrum of P-14a.

(53) FIG. 52 is a .sup.1H NMR spectrum of P-(14:1/1:1)a.

(54) FIG. 53 is a .sup.1H NMR spectrum of P-(14:1/1:3)a.

(55) FIG. 54 is a .sup.1H NMR spectrum of P-(14:21/1:1)a.

(56) FIG. 55 is a .sup.1H NMR spectrum of P-(14:7/1:1)a.

(57) FIG. 56 is a .sup.1H NMR spectrum of P-(14:7/1:3)a.

(58) FIG. 57 is a .sup.1H NMR spectrum of P-17a.

(59) FIG. 58 is a .sup.1H NMR spectrum of P-18a.

(60) FIG. 59 is a .sup.1H NMR spectrum of P-(18:1/1:1)a.

(61) FIG. 60 is a .sup.1H NMR spectrum of P-19a.

(62) FIG. 61 is a .sup.1H NMR spectrum of S-5a.

(63) FIG. 62 is a .sup.1H NMR spectrum of S-5b.

(64) FIG. 63 is a .sup.1H NMR spectrum of S-5c.

(65) FIG. 64 is a .sup.1H NMR spectrum of S-5d.

(66) FIG. 65 is a .sup.1H NMR spectrum of S-5e.

(67) FIG. 66 is a .sup.1H NMR spectrum of S-5f.

(68) FIG. 67 is a .sup.1H NMR spectrum of S-5g.

(69) FIG. 68 is a .sup.1H NMR spectrum of S-5h.

(70) FIG. 69 is a .sup.1H NMR spectrum of S-1f.

(71) FIG. 70 is a .sup.1H NMR spectrum of S-1g.

(72) FIG. 71 is a .sup.1H NMR spectrum of S-1h.

(73) FIG. 72 is a .sup.1H NMR spectrum of S-3f.

(74) FIG. 73 is a .sup.1H NMR spectrum of S-3g.

(75) FIG. 74 is a .sup.1H NMR spectrum of S-3h.

(76) FIG. 75 is a .sup.1H NMR spectrum of S-4f.

(77) FIG. 76 is a .sup.1H NMR spectrum of S-4g.

(78) FIG. 77 is a .sup.1H NMR spectrum of S-4h.

(79) FIG. 78 is a .sup.1H NMR spectrum of S-6f.

(80) FIG. 79 is a .sup.1H NMR spectrum of S-6g.

(81) FIG. 80 is a .sup.1H NMR spectrum of S-6h.

(82) FIG. 81 is a .sup.1H NMR spectrum of S-9g.

(83) FIG. 82 is a .sup.1H NMR spectrum of S-13g.

(84) FIG. 83 is a .sup.1H NMR spectrum of S-7g.

(85) FIG. 84 is a .sup.1H NMR spectrum of S-10g.

(86) FIG. 85 is a .sup.1H NMR spectrum of S-12g.

(87) FIG. 86 refers to a plot of viable microbe CFU after 18 hours (bacteria) or 42 hours (fungi) treatment at various polymer concentrations of P-1a-1 (6037B) and P-5c-1 (6050 D) (0, ½ MIC, MIC and 2×MIC). (a) S. aureus, (b) E. coli, (c) P. aeruginosa and (d) C. albicans.

(88) FIG. 87 is a graph showing viable microbes CFU after incubation with P-9a for 18 hours (bacteria) or 42 hours (fungi) at various concentrations (0, ½×MIC, 1×MIC and 2×MIC) (a) S. aureus, (b) E. coli, (c) P. aeruginosa and (d) C. albicans.

(89) FIG. 88 is a graph showing viable microbes CFU after incubation with P-7a for 30 seconds at various concentrations (0, 1×MIC, 2×MIC, 4×MIC, 8×MIC and 16×MIC) (a) S. aureus, (b) E. coli, (c) P. aeruginosa and (d) C. albicans.

(90) FIG. 89 is a graph showing viable microbes CFU after incubation with P-(1:7/1:1)a for 30 seconds at various concentrations (0, 1×MIC, 2×MIC, 4×MIC, 8×MIC and 16×MIC) (a) S. aureus, (b) E. coli, (c) P. aeruginosa and (d) C. albicans.

(91) FIG. 90 is a graph showing viable microbes CFU after incubation with P-1a-1 for 30 seconds at various concentrations (0, 1×MIC, 2×MIC, 4×MIC, 8×MIC and 16×MIC) (a) S. aureus, (b) E. coli, (c) P. aeruginosa and (d) C. albicans.

(92) FIG. 91 is a graph showing the percentage kill (%) of respective microbial strains (A: S. aureus, B: E. coli, C: P. aeruginosa and D: C. albicans) after incubation with surfactant S-3g for 18 hours (bacteria) or 42 hours (fungi).

(93) FIG. 92 is a graph showing the percentage kill (%) of respective microbial strains (A: S. aureus, B: E. coli, C: P. aeruginosa and D: C. albicans) after 30 seconds (solid) and 2 minutes (striped) of mixing with S-3g for concentrations of 4×, 8×, 16×, 32× and 64×MIC.

(94) FIG. 93 is a graph showing the percentage biofilm cell viability (A) and biomass (B) of A: S. aureus and biofilm cell viability (C) and biomass (D) of E. coli after treatment with surfactant S-3g for 18 hours.

(95) FIG. 95 is a graph showing the survival of K. pneumoniae 8637-infected mice without treatment (control) or treated with the polymer P-1a-1 (6037B) and imipenem. n=10.

(96) FIG. 95 is a graph showing bacterial counts in the blood and major organs of K. pneumoniae 8637-infected mice without treatment (control) or treated with the polymer P-1a-1 (6037B) and imipenem. n=3. (*, p<0.05; **, p<0.01; ***, p<0.001).

EXAMPLES

(97) Non-limiting examples of the disclosure will be further described in greater detail by reference to specific Examples, which should not be construed as in any way limiting the scope of the invention.

Example 1: Materials and Methods

(98) Materials

(99) All chemical reagents were purchased from Sigma-Aldrich (USA) or Tokyo Chemical Industry (Japan) and used as received unless specified. Mueller Hinton Broth (MHB) powder was purchased from BD Diagnostics (France) and used to prepare the microbial broths according to the manufacturer's instruction. Hydroxypropyl methyl cellulose (HPMC) H3785 and H7509, Igepal CO-520 and sodium dodecyl sulfate (SDS) were bought from Sigma-Aldrich. Igepal CA-630 was bought from MP Biochemicals Inc. (USA). N,N-bis(2-hydroxyethyl)dodecanamide (HDA) and N-decyl-b-D-glucopyranoside (DGP) were obtained from Flurochem. (UK). Cell lines of S. aureus (ATCC No. 6538), E. coli (ATCC No. 25922), P. aeruginosa (ATCC No. 9027), and C. albicans (ATCC No 10231) were obtained from ATCC, U.S.A., and reconstituted according to the suggested protocols. Rat red blood cells (rRBCs) were obtained from the Animal Handling Unit of Biomedical Research Centre, Singapore.

(100) Methods

(101) .sup.1H NMR Spectroscopy

(102) .sup.1H-NMR spectra using a Bruker Avance 400 spectrometer: operated at 400 MHz with the solvent proton signal as the internal reference standard.

(103) Size Exclusion Chromatography (SEC)

(104) Aqueous size exclusion chromatography (SEC) was conducted in the following solvent mixture: HPLC H2O:methanol:acetic acid=54:23:23 with 0.5 M sodium acetate (salt concentration with respect to entire solvent mixture) as the eluent at 0.5 mL/min flowrate. SEC was recorded on a Waters 2695 separation module equipped with a Waters 2414 differential refractometer and Waters Ultrahydrogel 120 and 500 columns (7.8×300 mm). Polymer solutions were prepared at 5 mg/mL and injection volume was 100 μL. Empower 3 software (Waters Corporation, U.S.A.) was used for data collection and analysis. The columns were calibrated with a combination of poly(ethylene glycol) and poly(ethylene oxide) standards (PSS Polymer Standard Service, GmbH, Germany).

(105) MIC Measurement

(106) Bacterial and fungal samples were inoculated according to ATCC's requirement. Briefly, bacteria and fungi were grown in Mueller Hinton Broth (MHB) at 37° C. and room temperature, respectively overnight to allow the microbes to enter the log growth phase with constant 100 rpm shaking. Minimum inhibitory concentration (MIC) of the polymers was determined using the broth microdilution method. 100 μL of 20% v/v water-containing broth with various polymer or Gemini-surfactant concentrations was first added into each well of a 96-well tissue culture plate. The inoculated overnight microbe solution was first diluted to an optical density (O.D.) reading of 0.07 using a microplate reader (TECAN, Switzerland). This will correspond to 3×10.sup.8 CFU/mL based on the McFarland 1 standard solution (CFU=colony forming units). The microbe solution was further diluted 1000 times to achieve a final 3×10.sup.5 CFU/mL. To each well, an equal volume of diluted microbes solution was added and incubated at 37° C. for 18 hours for bacteria samples and at room temperature for 42 hours for fungi samples. There were 6 replicates for each concentration tested, with negative control being microbes added into broth containing 20% v/v deionized water. MIC was taken as the lowest concentration at which there was no observable microbe growth when measured by the microplate reader.

(107) Killing Efficiency Test

(108) After MIC testing, all solutions from the control, ½MIC, MIC and 2MIC wells were collected and subjected to a series of ten-fold dilution. 20 μL of the diluted microbe solution was streaked onto pre-casted agar plates using LB+1.5% agar from the first Base. Agar plates with bacteria samples were incubated at 37° C. for 18 hours while those with fungi samples were incubated for 42 hours at room temperature before the CFUs were counted.

(109) Killing Kinetics

(110) Each polymer was diluted to 2, 4, 8 and 16×MIC with Mueller Hinton Broth (MHB) and distilled water. 600 μL of 3×10.sup.5 CFU/mL microbial suspension was added to each concentration of the polymer. After 30 seconds, the solution of polymer and microbes are serial diluted by folds of ten. 20 μL of the diluted microbial suspension was then plated onto agar plates and incubated for 24 hours and 42 hours at 37° C. and room temperature for bacteria and fungi respectively, then manually counted for CFU.

(111) Biofilm Culturing

(112) The bacteria stock was diluted in Mueller Hinton Broth (MHB) to get an optical density (OD) reading of 0.07 at 600 nm. 48 wells of a 96 flat-well plate were plated with 100 μL of bacteria suspension each. The plate was then incubated at 37° C. Every day over the next 7 days, MHB was carefully removed from the plate so as not to disturb the biofilm at the bottom of the well, and replaced with 100 μL of fresh MHB to each well.

(113) XTT Assay for Cell Viability of Biofilms

(114) After the biofilm was cultured, at the end of 7 days, MHB was removed from the wells. The biofilms were washed carefully with PBS 3 times to remove planktonic cells. The surfactant was weighed in a micro-centrifuge tube, and the weighed sample was diluted to 10,000 mg L.sup.−1 using distilled water. Serial dilution was conducted to obtain surfactant solutions having concentrations of 100 mg L.sup.−1, 70 mg L.sup.−1, 50 mg L.sup.−1, 35 mg L.sup.−1, 25 mg L.sup.−1, 20 mg L.sup.−1, and 10 mg L.sup.−1. The solvent was 20% distilled water and 80% MHB for each solution. The control was 20% distilled water and 80% MHB without surfactant. The surfactant solutions of different concentrations were added to the wells, 100 μL into each, with 6 replicates for each concentration. The plate was then incubated at 37° C. for 18 hours. Diluted XTT solution of 200 μL of PBS, 20 μL of XTT solution (1 mg mL) and 4 μL of menadione solution (1 mM) was added into each well. At the end of the incubation period, the surfactant solution was removed and 100 μL of XTT solution was added to each well. The plate was incubated at 37° C. in the dark for 3 hours. At the end of the incubation period, the optical density (OD) reading of the plate was taken at 490 nm.

(115) Biomass Assay

(116) After the biofilm was cultured, at the end of 7 days, MHB was removed from the wells. The biofilms were washed carefully with PBS 3 times to remove planktonic cells. The surfactant was weighed in a micro-centrifuge tube, and the weighed sample was diluted to 10000 mg L.sup.−1 using distilled water. Serial dilution was conducted to obtain surfactant solutions of concentrations 100 mg L.sup.−1, 70 mg L.sup.−1, 50 mg L.sup.−1, 35 mg L.sup.−1, 25 mg L.sup.−1, 20 mg L.sup.−1, and 10 mg L.sup.−1. The solvent was 20% distilled water and 80% MHB for each solution. The control was 20% distilled water and 80% MHB without surfactant. The surfactant solutions of different concentrations were added to the wells, 100 μL into each, with 6 replicates for each concentration. The plate was incubated at 37° C. for 18 hours. At the end of the incubation period, the surfactant solution was removed and 100 μL of 0.1% safranin O solution was added to each well. The plate was incubated at 37° C. in the dark for 15 minutes. At the end of the incubation period, the excess stain was removed by washing at least three times with PBS. Ethanol (200 μL of 70% solution) was then added to each well. The plate was incubated at 37° C. in the dark for 15 minutes. At the end of the incubation period, the optical density (OD) reading of the plate was taken at 550 nm.

(117) Hemolysis Assay

(118) Freshly obtained rat red blood cells (rRBC) from the Animal Handling Unit of Biomedical Research Center (Singapore) was used to test the hemolytic activity of the polymers and Gemini-surfactants. The rRBC was diluted to 4% v/v with phosphate-buffered saline (PBS) and added to an equal volume of 0.2% v/v Triton-X solution. A microplate reader was used to measure the optical density (O.D.) reading to make sure the value is between 0.5 and 0.6. An equal volume of different polymer or Gemini-surfactant concentrations was added to the diluted blood and incubated for 1 hour at 37° C. The samples were centrifuged at 1000 g at 4° C. for 5 minutes and 100 μL of the supernatant was collected and transferred to a 96-well tissue culture plate. There were 4 replicates for each concentration. Haemoglobin release was measured using the microplate reader at 576 nm. rRBCs treated with PBS were used as the positive control, while those treated with 0.2% v/v Triton-X was used as the negative control.

(119) Percentage of hemolysis was calculated as follows:
Hemolysis (%)═[(O.D..sub.576 nm of the treated sample−O.D..sub.576 nm of negative control)/(O.D..sub.576 nm of positive control−O.D..sub.576 nm of negative control)]×100%

(120) HC.sub.50 was taken as the polymer concentration at which the polymer causes 50% hemolysis.

(121) Bacterial Strain and Culture Conditions for In Vivoassay

(122) Clinically isolated multidrug-resistant K. pneumoniae strain was extracted from a patient's phlegm and provided by The First Affiliated Hospital of Medical College, Zhejiang University (Hangzhou, China). The isolate was identified by routine laboratory methods and stored in 20% (v/v) glycerol at

(123) −80° C. Bacteria were grown in Mueller-Hinton (MH) agar plate at 37° C. prior to use. Animals

(124) ICR mice (female, 7 weeks old, 24-26 mg) were used for the in vivo studies. Immunosuppression was induced by intraperitoneal injection of 200 mg/Kg cyclophosphamide (Hengrui Corp, Jiangsu Province, P. R. China) 4 days prior to infection. Mice were anesthetized by intra-peritoneal injection of 1% pentobarbital (40 mg/kg, Sigma).

(125) In Vivo Toxicity Study

(126) To assess the acute systemic toxicity of P-1a-1 (6037B) to the mice, the median lethal dose (LD.sub.50) was determined. Specifically, the mice were randomly assigned into six treatment groups (six mice per group). After dissolution in phosphate buffered serum (PBS), P-1a-1 (6037B) was administered intraperitoneally at designated doses (i.e. 20.0, 30.0, 45.0, 67.5, 101.3, and 151.9 mg/kg, 0.2 mL/20 g). The number of surviving mice in each group was monitored for five days after treatment, and the values of LD.sub.50 were calculated by a regression technique such as the BLISS method. Such method may involve the use of software BLISS LD50, which allows one to obtain the LD.sub.50 upon input of the relevant data to the software. Other suitable regression techniques may also be used as appropriate.

(127) Pulmonary Infection

(128) To investigate the in vivo antibacterial efficacy of P-1a-1 (6037B), a pneumonia mouse model was established. The immunosuppression of mice was induced as described above. Overnight cultures of K. pneumoniae 8637 were harvested and suspended in PBS. Before instillation with K. pneumoniae 8637, the mice were anesthetized using an intraperitoneal injection with 1% pentobarbital (40 mg/kg). Each of the immunosuppressed mice was infected intranasally with 0.03 mL of the bacterial suspension at designated doses (i.e. 1×10.sup.9, 1.5×10.sup.9, 2.3×10.sup.9, 3.5×10.sup.9, 5.3×10.sup.9, 8.0×10.sup.9 CFU/ml, four mice per group). The minimum lethal dose was defined as the lowest dose, which was sufficient to cause 100% mortality. It was determined from the survival rate of mice at 5 days post-infection by the BLISS method.

(129) Efficacy of P-1a-1 (6037B) in Treating K. pneumoniae 8637-Induced Pneumonia

(130) The bacterial suspension having the minimum lethal dose (0.03 mL/mice) was introduced to mice intranasally, after which P-1a-1 (6037B) and imipenem (a clinically used antibiotic for treatment of Gram-negative bacterial infections) were administered intraperitoneally once daily for 3 days starting at 4 hours after infection at designated doses (i.e. 0.1, 0.5, 1.0, 2.0, 4.0, 8.0 mg/kg for P-1a-1 (6037B), 0.1, 1.0, 5.0, 10.0, 20.0, 40.0 mg/kg for imipenem, 0.2 mL/20 g, four mice per group). The number of surviving mice in each group was recorded for 5 days to assess ED.sub.50 by the BLISS method.

(131) To further determine the in vivo therapeutic efficacy of P-1a-1 (6037B), survival of the K. pneumoniae 8637-infected mice was monitored with and without treatment. Briefly, the mice were randomly divided into a control group, a P-1a-1 (6037B)-treated group and an imipenem-treated group (ten mice per group). After being anesthetized, each of the immunosuppressed mice was inoculated intranasally with 0.03 mL of bacterial suspension at the minimum lethal dose determined above. Then, P-1a-1 (6037B) and imipenem (i.e. 2.0 mg/kg for imipenem, 1.0 mg/kg for P-1a-1 (6037B)) were administered intraperitoneally once daily for 3 consecutive days starting at 4 hours after infection. The mice were monitored for a period of five days, and the number of surviving mice in each group was recorded. Survival was expressed using the Kaplan-Meier curve.

(132) Lung, blood, liver, spleen and kidney samples were obtained to assess bacterial counts. Briefly, after anesthesia with 40 mg/kg pentobarbital, immunosuppressed mice were instilled intra-nasally with 2×10.sup.7 (three mice/group) colony-forming units (CFUs) of K. pneumoniae 8637. The polymer sample P-1a-1 (6037B) and imipenem at ED95 doses (0.2 mL/20g) were administered intraperitoneally once daily for 3 consecutive days starting at 4 hours after infection. At five days post-infection, the count of bacteria in the lung, blood, liver, spleen and kidney were determined by Mueller-Hinton (MH) agar plating. The results were shown as mean±SD.

Example 2: Synthesis

General Procedure for the Degradable d-A.SUB.2 .Monomer

Method A: Monomers Containing Aromatic Esters and Amides

Representative Example—Synthesis of N, N′-((ethane-1,2-diylbis(oxy)bis(ethane-2,1-diyl))bis(4-(chloromethyl) benzamide) (d-A.SUB.2.-4)

(133) In a round two-necked bottom flask (500 mL) equipped with a magnetic stir bar and nitrogen inlet adaptor, 4-(chloromethyl)benzoyl chloride (10.55 g, 55.81 mmol, 2.0 equiv.) and tetrahydrofuran (THF, 20 mL) were allowed to equilibrate under ice-cold conditions for about 30 minutes. To this solution, a mixture of 2,2′-(ethane-1,2-diylbis(oxy))bis(ethan-1-amine) (4.14 g, 27.94 mmol, 1.0 equiv.) and triethylamine (12.0 mL, 8.7 g, 86.1 mmol, 3.1 equiv.), dissolved in THF (40 mL) were added dropwise via a dropping funnel over approximately 30 minutes. A white precipitate formed immediately. The reaction was allowed to proceed at room temperature for an additional 90 minutes. Deionized water (about 200 mL) was added to the reaction mixture to dissolve the triethylamine salts and also precipitate the product. This suspension was further chilled in an ice bath for about an hour, followed by the isolation of the products as a solid by vacuum filtration. The product was washed with deionized water and was dried under high vacuum to yield a white powdery solid (11.3 g, 89.3%).

(134) Monomers d-A.sub.2-1 to 5 and 12 were prepared by this method. Typical yields˜70-94%. N, N′-(ethane-1,2-diyl)bis(4-(chloromethyl)benzamide) (d-A.sub.2-1) N, N′-(1,4-phenylene)bis(4-(chloromethyl)benzamide) (d-A.sub.2-2) N, N′-(hexane-1,6-diyl)bis(4-(chloromethyl)benzamide) (d-A.sub.2-3) N,N′-(1,3-phenylenebis(methylene))bis(4-(chloromethyl)benzamide) (d-A.sub.2-4) N, N′-((ethane-1,2-diylbis(oxy))bis(ethane-2,1-diyl))bis(4-(chloromethyl) benzamide) (d-A.sub.2-5)

Method B

Representative Example—Synthesis of N,N′-(((oxybis(ethane-2,1-diyl))bis(oxy))bis (propane-3,1-diyl)) bis(4-(chloromethyl) benzamide (d-A.SUB.2.-6)

(135) A protocol similar to Method A was used. However, d-A.sub.2-6 did not precipitate upon addition of water (about 100 mL). Instead, an oily layer separated. The product was first extracted from the reaction mixture with ethyl acetate (3×200 mL). The combined organic layers were dried with sodium sulphate and the volatiles were removed to result in pale yellow crude product, which was further purified by recrystallizing from a minimal amount of ethyl acetate. Isolated yield: 10.16 g (65.2% crop 1—recrystallized from ethyl acetate).

(136) Monomers d-A.sub.2-6—11, 13, 19 and 20 were prepared by Method B. Typical yields˜55-85%. If the crude product by thin layer chromatography was not a single spot, the monomer was further purified by recrystallization or flash column chromatography using hexane/ethyl acetate or chloroform/ethylacetate combination. In general, for the di-ester series, a protocol, similar to that of d-A.sub.2-6 synthesis was followed (Method B). Typically reaction was conducted overnight.

Method C: Monomers Containing Aliphatic Esters and Amides

Representative Example—Synthesis of N,N′-(ethane-(1,2-diyl)bis(4-chlorobutanamide) (d-A.SUB.2.-4)

(137) In a two-necked round bottom flask (250 mL) equipped with magnetic stir bar and nitrogen inlet adaptor, 4-Chlorobutyryl chloride (11.4 mL, 14.2 g, 100.7 mmol, 2.02 equiv.) and tetrahydrofuran (THF, 25 mL) were allowed to equilibrate under ice-cold conditions for about 30 minutes. To this solution, a mixture of ethylene diamine (3.00 g, 49.8 mmol, 1.0 equiv.) and triethylamine (15.0 mL, 10.9 g, 107.7 mmol, 2.16 equiv.), dissolved in THF (75 mL) were added drop-wise via dropping funnel over ˜30 minutes. White precipitates were formed immediately. The reaction mixture was allowed to proceed at room temperature for additional 90 minutes. DI water (˜100 mL) was added to the reaction mixture to dissolve the triethylamine salts and the product was extracted into ethylacetate (150 mL). Organic layer was washed with 2×1N HCl (100 mL), 2×Satd. NaHCO.sub.3 aqueous solution and 1×Satd. NaCl aqueous solution and dried over Na.sub.2SO.sub.4 and concentrated in vacuuo to yield a pale yellow solid as the crude product. Crude product was recrystallized from hot acetonitrile and the crystalline solid was washed with ethylacetate:hexane (1:1 v/v) mixture. The product was dried under high vacuum to yield white crystalline solid (4.42 g, ˜33%).

(138) Monomers d-A.sub.2-15 and 16 were prepared by this method. However the crude product was purified by flash column chromatography using hexane/ethyl acetate solvent mixtures. Monomers d-A.sub.2-18 was also prepared by this method. However succinyl chloride and iodoethanol were used instead. Typical yields˜50-70%.

Method D: Monomers Containing Aliphatic Esters with Alkyl Iodides, Obtained Via Standard Finkelstein Reaction

Representative Example—Synthesis of ethane-1,2-diyl bis(4-iodobutanoate) (d-A.SUB.2.-17)

(139) In a round bottom flask (50 mL) equipped with magnetic stir bar, condenser (with circulating cooling water) and nitrogen inlet adaptor, ethane-1,2-diyl bis(4-chlorobutanoate) (d-A.sub.2-16 2.71 g, 10 mmol, 1.0 equiv.), sodium iodide (5.12 g, 34.2 mmol, 3.4 equiv.) and acetone (30 mL) were added and the reaction mixture was allowed to proceed for 48 hours at refluxing conditions by maintaining the oil bath at 60° C. Then, insolubles were removed by filtration and solvent removed under vaccuo to result in crude product, which was further purified by flash column chromatography to result in target compound as an oil (3.46 g 76.2%).

General Procedure for the Synthesis of Degradable Polyionenes Via Addition Route

Method A

(140) Unless otherwise noted, Method A was the standard method used for the synthesis of polyionenes from monomers containing benzylchloride derivatives

Representative Example—Synthesis of poly(d-A.SUB.2.-1-co-tetramethyl-1,3-diaminopropane), P-1a

(141) In a scintillation vial (20 mL) equipped with a magnetic stir bar, monomer d-A.sub.2-1 (A.sub.2-type, 360.0 mg, 985.6 μmol) and tetramethyl-1,3-diaminopropane (B.sub.2-type, 131.0 mg, 1006 μmol) were suspended in dimethylformamide (DMF) (3.0 mL). The reaction mixture was gently heated with a heat gun to render a clear solution. The clear solution was allowed to stir at room temperature. The reaction mixture tuned cloudy and at about 1.5 hours, a polymer was found to deposit on the walls of the vial. The reaction mixture was allowed to stir further overnight, and was precipitated into diethyl ether (50 mL) to result in a solid white powder. The white power was dissolved/suspended in about 5 mL methanol, then about 45 mL diethylether was slowly added to precipitate the polymer and this process was repeated once more. The solids were isolated and dried in vacuuo to result in a white solid at near quantitative yields. The product was further purified by first dissolving in deionized water, followed by extensive dialysis against deionized water using a dialysis membrane (MWCO=1 kDa), and lyophilization to result in the target polymer.

(142) For the synthesis of copolymers, the combined molar ratio of different d-A.sub.2 and/or A.sub.2 to B.sub.2 monomers were kept at 1:1.

Method B: Copolymerization of Monomers Containing Aliphatic Chlorides

Representative Example—Synthesis of poly(d-A.SUB.2.-14-co-tetramethyl-1,3-diaminopropane), P-14a

(143) In a scintillation vial (20 mL) equipped with magnetic stir bar, monomer d-A.sub.2-14 (A.sub.2-type, 539.2 mg, 2003 μmol) and tetramethyl-1,3-diaminopropane (B.sub.2-type, 260.8 mg, 2003 μmol) were suspended in DMF (3.0 mL). The reaction mixture was allowed to stir at 60° C. overnight (˜18h), and was then precipitated into diethyl ether (50 mL) to result in a viscous semi-solid. The solid was dissolved/suspended in 5 mL methanol, and 45 mL diethylether was slowly added to precipitate the polymer and this process was repeated once more. The solids were isolated and dried in vacuuo to result in white solid. Polymer was further purified by first dissolving in DI water, followed by extensive dialysis against DI water using dialysis membrane (MWCO=1 kDa), and lyophilization to result in target polymer.

Method C: Copolymerization of Monomers Containing Benzylchloride Derivatives and Aliphatic Chlorides

Representative Example—Synthesis of poly(d-A.SUB.2.-14-co-tetramethyl-1,3-diaminopropane/d-A2-1-co-tetramethyl-1,3-diaminopropane), P-(14:1/1:1)a

(144) In a scintillation vial (20 mL) equipped with magnetic stir bar, monomer d-A.sub.2-14 (A.sub.2-type, 270.2 mg, 1004 μmol) and tetramethyl-1,3-diaminopropane (B.sub.2-type, 260.9 mg, 2003 μmol) were suspended in DMF (3.0 mL). The reaction mixture was allowed to stir at 60° C. overnight (˜18h). To this reaction mixture comonomer d-A.sub.2-1 (A.sub.2-type, 365.2 mg, 1000 μmol) along with additional DMF (3.0 mL) was added. The reaction was allowed to proceed for another 2 hours at 60° C. and was then precipitated into diethyl ether (2×50 mL). Precipitates were dissolved/suspended in ˜5 mL methanol, and ˜45 mL diethylether was slowly added to precipitate the polymer and this process was repeated once more. The solids were isolated and dried in vacuuo to result in solid. Polymer was further purified by first dissolving in DI water, followed by extensive dialysis against DI water using dialysis membrane (MWCO=1 kDa), and lyophilization to result in target polymer.

Method D: Copolymerization of Monomers Containing Aliphatic Iodide

Representative Example—Synthesis of poly(A.SUB.2.-18-co-tetramethyl-1,3-diaminopropane), P-18a

(145) In a scintillation vial (20 mL) equipped with magnetic stir bar, monomer d-A.sub.2-18 (A.sub.2-type, 428.5 mg, 1006 μmol) and tetramethyl-1,3-diaminopropane (B.sub.2-type, 130.6 mg, 1003 μmol) were suspended in DMF (1.5 mL). The reaction mixture was allowed to stir at room temperature overnight (˜18h), and was then precipitated into diethyl ether (50 mL) to result in a viscous semi-solid. The solid was dissolved/suspended in ˜2-3 mL methanol, and ˜45 mL diethylether was slowly added to precipitate the polymer and this process was repeated once more. The solids were isolated and dried in vacuuo to result in white solid. Polymer was further purified by first dissolving in DI water, followed by extensive dialysis against DI water using dialysis membrane (MWCO=1 kDa), and lyophilization to result in target polymer.

Method E: Copolymerization of Monomers Containing Benzylchloride Derivatives and Aliphatic Iodides

Representative Example; Synthesis of poly(d-A.SUB.2.-18-co-tetramethyl-1,3-diaminopropane/d-A.SUB.2.-1-co-tetramethyl-1,3-diaminopropane), P-(18:1/1:1)a

(146) In a scintillation vial (20 mL) equipped with magnetic stir bar, monomer d-A.sub.2-18 (A.sub.2-type, 211.4 mg, 496 μmol) and tetramethyl-1,3-diaminopropane (B.sub.2-type, 130.6 mg, 1003 μmol) were suspended in DMF (1.5 mL). The reaction mixture was allowed to stir at room temperature overnight (˜18h). To this reaction mixture comonomer d-A.sub.2-1 (A.sub.2-type, 185.4 mg, 508 μmol) along with additional DMF (1.5 mL) was added and the reaction mixture was allowed to stir at room temperature additional 18h, followed by precipitation into diethyl ether (50 mL) to result in a viscous semi-solid. The solid was dissolved/suspended in ˜2-3 mL methanol, and ˜45 mL diethylether was slowly added to precipitate the polymer and this process was repeated once more. The solids were isolated and dried in vacuuo to result in white solid. Polymer was further purified by first dissolving in DI water, followed by extensive dialysis against DI water using dialysis membrane (MWCO=1 kDa), and lyophilization to result in target polymer.

General Procedure for the Synthesis of Degradable Gemini-Surfactants

Representative Example—Synthesis of S-1g

(147) In a scintillation vial (20 mL) equipped with a magnetic stir bar, monomer d-A.sub.2-1 (A.sub.2-type, 369.3 mg, 1010.9 μmol) and N,N-dimethyldodecylamine (g, B-type, 1200 μL, 944 mg, 4424 μmol) were suspended in dimethylformamide (DMF) (2.0 mL). The reaction mixture was gently heated with a heat gun to render a clear solution. The clear solution was allowed to stir at room temperature overnight. The reaction mixture was then precipitated into diethyl ether (50 mL) to result in a solid. The solid was dissolved/suspended in about 5 mL methanol, then about 45 mL diethylether was slowly added to precipitate the surfactant. This process was repeated once more. The solids were isolated and dried in vacuuo to result in a white solid.

(148) Preparation of Surgical Scrub Formulation

(149) Polymer P-1a-1 (6037B) was diluted with de-ionized (DI) water to obtain a 5 wt % stock. Igepal 630, hydroxpropyl methylcellulose (HPMC) 3785 and either N,N-bis(2-hydroxyethyl)dodecanamide (HDA) or N-decyl-b-D-glucopyranoside (DGP) was prepared at a ratio of 5:1:5 wt %. 100 μL of P-1a-1 (6037B) and 900 μL of DI water were added to the rest of the mixture. The mixture was sonicated at room temperature for 5 minutes before mixing by vortexing. This was repeated for another 2 more times. The tubes were placed in 4° C. overnight to ensure that all the HPMC was totally dissolved. 0.1 mL of each formulation (3 replicates) was added into each well in a 96-well plate before 100 μL of bacteria suspension was added. Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa obtained from ATCC were reconstituted from its lyophilized form according to the manufacturer's protocol. Microbial samples were cultured in MHB at 37° C. under constant shaking of 300 rpm. Prior to mixing, the microbial sample was first inoculated overnight to enter its log growth phase. The concentration of microbial solution was adjusted to give an initial optical density (O.D.) reading of approximately 0.07 at 600 nm wavelength on microplate reader (TECAN, Switzerland), which corresponds to the concentration of McFarland Standard No. 1 (3×10.sup.8 CFU ml.sup.−1), the microbial solution was further diluted by 1000 to achieve an initial loading of 3×10.sup.1 CFU ml.sup.−1. Three different incubation times (10 sec, 30 sec and 10 minutes) were tested for each formulation. The microbial samples were taken out from each well for a series of 10-fold dilutions. 20 μL of the diluted microbial solution was streaked onto an agar plate (LB Agar from first Base). The plates were incubated for 24 hours at 37° C. and counted for colony-forming units (CFU).

Example 3: 1H NMR Characterization of the Synthesized Compounds

(150) The synthesized compounds were characterized by .sup.1H NMR spectroscopy. The .sup.1H NMR spectra for the respective compounds are in the Figures as outlined in Table 1 below:

(151) TABLE-US-00001 TABLE 1 .sup.1H NMR spectra of the synthesized compounds Compound FIG. d-A2-1 FIG. 3 d-A2-2 FIG. 4 d-A2-3 FIG. 5 d-A2-4 FIG. 6 d-A2-5 FIG. 7 d-A2-6 FIG. 8 d-A2-7 FIG. 9 d-A2-8 FIG. 10 d-A2-9 FIG. 11 d-A2-10 FIG. 12 d-A2-11 FIG. 13 d-A2-12 FIG. 14 d-A2-13 FIG. 15 d-A2-14 FIG. 16 d-A2-15 FIG. 17 d-A2-16 FIG. 18 d-A2-17 FIG. 19 d-A2-18 FIG. 20 d-A2-19 FIG. 21 d-A2-20 FIG. 22 P-1a-1 FIG. 23 P-1c-1 FIG. 24 P-3a FIG. 25 P-3c FIG. 26 P-5c-1 FIG. 27 P-6a FIG. 28 P-6b FIG. 29 P-6c FIG. 30 P-4a FIG. 31 P-4b FIG. 32 P-4c FIG. 33 P-1b FIG. 34 P-5a FIG. 35 P-5b FIG. 36 P-21a FIG. 37 P-7a FIG. 38 P-(7:1/1:1)a FIG. 39 P-(7:1/3:7)a FIG. 40 P-(7:1/1:9)a FIG. 41 P-8a FIG. 42 P-8b FIG. 43 P-8c FIG. 44 P-(8:21/1:1)a FIG. 45 P-(8:21/1:9)a FIG. 46 P-9a FIG. 47 P-10a FIG. 48 P-12a FIG. 49 P-13a FIG. 50 P-14a FIG. 51 P-(14:1/1:1)a FIG. 52 P-(14:1/1:3)a FIG. 53 P-(14:21/1:1)a FIG. 54 P-(14:7/1:1)a FIG. 55 P-(14:7/1:3)a FIG. 56 P-17a FIG. 57 P-18a FIG. 58 P-(18:1/1:1)a FIG. 59 P-19a FIG. 60 S-5a FIG. 61 S-5b FIG. 62 S-5c FIG. 63 S-5d FIG. 64 S-5e FIG. 65 S-5f FIG. 66 S-5g FIG. 67 S-5h FIG. 68 S-1f FIG. 69 S-1g FIG. 70 S-1h FIG. 71 S-3f FIG. 72 S-3g FIG. 73 S-3h FIG. 74 S-4f FIG. 75 S-4g FIG. 76 S-4h FIG. 77 S-6f FIG. 78 S-6g FIG. 79 S-6h FIG. 80 S-9g FIG. 81 S-13g FIG. 82 S-7g FIG. 83 S-10g FIG. 84 S-12g FIG. 85

Example 3: Discussion of the Synthesis of (Poly)Ionenes

(152) Synthesis of d-A2 Monomers

(153) It was found that (poly)ionenes with tunable degradability can be developed from functional monomers with built-in cleavable bonds. To achieve this, first a robust and facile method to produce a functional bis(halide), comprising a cleavable bond, was developed. Incorporation of an amide bond as the cleavable linker would result in the requisite stability for the compounds that would translate to meaningful shelf-life while at the same time, introducing potential biodegradability to the resultant (poly)ionenes.

(154) A library of degradable bis(halide) A.sub.2-type monomers were synthesized by reacting two equivalence of chloromethyl benzoyl chloride with a diamine in the presence of a base, to result in d-A.sub.2 monomers 1 to 6, with good yields (FIG. 1 and FIG. 2). The selection of diamine would enable the tailoring of the hydrophobicity/hydrophilicity and/or the flexibility/rigidity, which are also determined by the spacer between the benzyl chloride functionalities. This degradable bis(halide) d-A.sub.2 monomer could be polymerized with commercially available B.sub.2-type bis(dimethylamino)-monomers to result in polyionenes P using dimethylformamide (DMF) as the solvent at room temperature.

(155) By replacing diamine with a diol or aminoalcohol, d-A.sub.2 monomers 7 to 13 could be synthesized. Readily available precursors enable systematic variation of the spacer lengths in these monomers. For example, monomers containing diesters were synthesized by increasing the spacer lengths from propyl to dodecyl (d-A.sub.2 monomers 7 to 11). Similarly with a fixed hexyl spacer, the specific linker chemistry has been varied from di-amides, to di-esters to amide-ester combination (d-A.sub.2 monomers 3, 9 and 13 respectively). By replacing chloromethyl benzoyl chloride with 4-chlorobutyryl chloride, monomers with aliphatic di-amides, diesters and amide-ester combinations were synthesized (d-A.sub.2 monomers 14 to 16).

(156) To modulate the reactivity of aliphatic halides, Finkelstein reaction can be conducted to exchange the chloride to bromide or iodide. To this end, d-A.sub.2 monomer 16 was subjected to halogen exchange reaction in the presence of sodium iodide to result in d-A.sub.2 monomer 17. With an aliphatic iodide, this monomer would enable higher reactivity so that subsequent polymerization and reactions can be conducted at a significantly lower temperature than the aliphatic chlorides. Alternatively, similar reactive monomers with aliphatic esters can also be directly synthesized from a corresponding iodoalcohol such as iodoethanol with a bis-acidchloride such as succinyl chloride to result in target d-A.sub.2 monomer 18. Functionalities can also be introduced along with the linker. As a proof-of-concept, a protected secondary amine was introduced via commercially available diethanol amine-based precursor or allyl side-chain functionality introduced via commercially available precursors to result in d-A.sub.2 monomer 19 and 20, respectively. The choice of difunctional spacer (diamine or diol or amino alcohols) can be used to tailor the hydrophobic/hydrophilic, flexibility/rigidity and degradability of the polymers.

(157) Synthesis of Polyionenes

(158) This degradable bis(halide) d-A.sub.2 monomers can be polymerized with commercially available B.sub.2-type bis(dimethylamino)-monomers to result in polyionenes P using DMF as the solvent. This reaction can proceed well at room temperature for monomers containing benzyl chloride-derivatives and alkyliodides. For monomers with alkylchlorides, polymerization was conducted at 60° C. The resulting polyionenes had degradable amides, esters or a combination of both amide and ester bonds along the backbone. Post polymerization, the reaction mixture was precipitated thrice into diethylether and the resultant solids were further purified by extensive dialysis against de-ionized (DI) water, followed by lyophilization to result in solids.

(159) These polymers are in general coded by three characters. For, example P-1a, denotes the polymer synthesized using d-A.sub.2 monomer 1 and B.sub.2 monomer a (FIG. 1, FIG. 2 and Table 2). Additionally monomers such as P-1a-n, indicate that this entry is from n.sup.th independently repeated synthesis. If a combination of two different monomers were used for the polymerization, then these polymers are coded as follows: P-(1:21/1:1)a; this polymer has been prepared by using a combination of two-different A2 monomers 1 and 21 (with an initial feed ratio of 1:1, respectively) together with B.sub.2 monomer a (FIG. 1, FIG. 2 and Table 2). It is important to note that the overall molar ratio of A.sub.2 to B.sub.2 was kept 1:1.

(160) Apart from having the ability to tailor the polymer composition from the choice of a single d-A.sub.2 monomer, these monomers can be successfully combined with other monomers comprising of different classes (such as di-amides and di-esters) in different ratios to modulate polymer properties. Moreover, commercially readily available A.sub.2 monomer 21, can also be well integrated onto these platforms. Addition polymerization of d-A.sub.2 monomers, irrespective of the nature of their linkers (di-amides or di-esters or a combination of ester and amide), benzyl chloride-derivatives and alkyliodides proceeded well with the B.sub.2 monomer at room temperature.

(161) In order to copolymerize the less-activated alkyl chloride-functionalized monomers with that of activated benzyl chloride monomers, a two-step approach has been used. For example, in entry 41 of Table 2, 0.5 equivalents of d-A.sub.2 monomer 14 is first reacted with 1.0 equiv. of B.sub.2 monomer at 60° C. for 18 hours. To this reaction mixture, the remaining equivalents of d-A.sub.2 monomer, (for this example, an additional 0.5 equivalents of d-A.sub.2 monomer 7) is added and the reaction is allowed to proceed for another 2 hours at 60° C. to result in a polymer that has not only the combination of both amide and ester linkers but also successful integration of monomers with different reactivities.

(162) In general, polyionenes were obtained with a molecular weight in the range of about 2 to 20 kDa and molar mass dispersity in the range of about 1.2 to 2.9. It is important to note that, the approach described here can be readily extended to degradable B.sub.2-type bis(dimethylamino)-containing monomers as well. Also by changing from a diamino linker to a diol linker, one could potentially change the nature of the degradable bond from amide to ester, impacting the rate of degradation. Installation of degradable moieties into polyionenes could lead to biocompatible and environmentally-degradable antimicrobials.

(163) The resulting polyionenes have degradable amide bonds along the backbone. Post polymerization, the reaction mixture was precipitated thrice into diethylether and the resultant solids were further purified by extensive dialysis against deionized (DI) water, followed by lyophilization to result in a fluffy solid.

(164) For the purposes of this disclosure, these polymers are coded by three characters. For, example P-1a, denotes the polymer synthesized using d-A.sub.2 monomer 1 and B.sub.2 monomer a (FIG. 2). Likewise, by using excess of the functional tertiary amines, under similar conditions to that of polymerization, a library of Gemini-surfactants S were readily accessed.

(165) After reaction, the surfactant was purified by precipitating thrice into diethylether, to remove DMF and excess tertiary amines, to result in a solid product. For the purposes of this disclosure, these surfactants are coded by three characters. For, example S-5a, denotes the surfactant synthesized using d-A.sub.2 monomer 5 and B tertiary amine a (FIG. 2). It is important to note that, the approach described here can be readily extended to degradable B.sub.2-type bis(dimethylamino)-containing monomers as well. In addition, by changing from a diamino linker to a diol linker, one could change the nature of the degradable bond from amide to ester, altering the rate of degradation.

Example 4: Antimicrobial and Hemolytic Activities of Polyionenes (P)

(166) Minimum inhibitory concentrations (MIC) of the (poly)ionenes were evaluated by a broth microdilution method against clinically relevant microorganisms such as S. aureus (Gram-positive bacteria), E. coli and P. aeruginosa (Gram-negative bacteria), and C. albicans (fungi). Also as an indication of toxicity of these (poly)ionenes, hemolytic assays were conducted to evaluate their hemolytic activity (HC.sub.50 values) against rat red blood cells (rRBCs).

(167) Remarkably, most of the polymers tested were shown to have relatively low MIC values of 2 to 31 ppm across different microbes and relatively high HC.sub.50 values of >1000 ppm (Table 2). Performance of polymer P-1a (Table 2, entry 1) was found to be optimal with extremely low MICs of 2 to 8 ppm and at the same time demonstrated very high HC.sub.50 values of >2000 ppm. These results were consistent across multiple batches, highlighting the reproducibility of these materials (Table 2, entries 1, 12, 18 and 22).

(168) By comparing different types of commercially available B.sub.2 monomer (a, b and c, FIG. 2), used as a comonomer with the addition polymerization of d-A.sub.2 monomers (FIG. 2, 1 to 6 and 8), in general there was no major change in the MIC values that could be attributed due to the change in B.sub.2 monomer (Table 2, entries 1-17 and 22-29).

(169) Among d-A.sub.2 monomers from di-ester series (FIG. 2, 7 to 12), it is evident that, in general, the shorter the spacer, the better the potency and selectivity (Table 2, entries 23, 27, 32-34). In this di-ester series, polymer P-8a was found to have the highest potency and selectivity. For a fixed spacer length of hexyl, by probing the role of the linker chemistry, in terms of di-amide versus di-ester versus amide-ester combination, it was found that although the potency was not affected, selectivity was (Table 2, entries 3, 32 and 35). HC.sub.50 values were highly dependent on the nature of linker chemistry and followed the following trend: P-3a>P-13a>>P-9a. This finding underscores the importance of the combination of amide and ester bonds in tailoring selectivity.

(170) The amide-ester combinations can be introduced not just from a well-defined monomer (d-A.sub.2 monomer 13 and 15), but also by copolymerizing different classes of monomers. Unlike monomers where the ratio of amide to ester is fixed at 1:1, copolymerization of di-ester and di-amide monomers offers the ability to better tailor the ratio of ester to amide. Apart from having an effect on selectivity, this approach can also improve the potency (Table 2, entries 23-26). By increasing the content of d-A.sub.2 monomer 1 in the polymerization of d-A.sub.2 monomer 7, it was found that the potency of these polymers was improved, across multiple strains. These findings are also supported by killing kinetics and MBC data (discussed further below).

(171) Ability to incorporate aliphatic amides and esters along with aromatic amides and/or esters can offer unprecedented opportunity to tailor biodegradation, owing to high selectivities of the enzymes that cleave these bonds. To this end, d-A.sub.2 monomers 14 to 18 were synthesized and selected examples were polymerized with other classes of monomers and their biological properties were evaluated (Table 2, entries 36-44). Collectively these results demonstrate that these monomers containing aliphatic amides or esters can be well integrated into other classes of monomers containing aromatic benzyl chloride-type derivatives. More importantly, these polymers have a broad spectrum of action and high selectivity.

(172) The polymers from d-A.sub.2 monomers 14 and 17 were found to be active against bacteria but not very potent against fungi (Table 2, entries 36 and 42). This can be advantageous for selective eradication of bacteria and for selectively growing commercially relevant fungi and hence may have implications in biotechnology, food and a multitude of other disciplines where such selective killing of microbes is necessary.

(173) Ability to copolymerize different sub-classes of d-A.sub.2 monomers and also with other commercially readily available, inexpensive A.sub.2 monomer (for example compound 21 of FIG. 2) to result in polymers that are potent and selective illustrates the versatility and practical relevance of this approach. This approach also offers unprecedented opportunity to tailor physical, chemical and biological properties of materials (Table 2, entries 19, 20, 24-26, 30, 31, 37-41 and 44). Moreover, the extent and speed of degradation of these polymers in the biological and environmental contexts can also be controlled.

(174) Additional functionalities can be introduced in this d-A.sub.2 monomer via the linkers as illustrated by monomer 19 and 20. Resultant polymers from some of these polymers were also found to be potent and their selectivity modulated through additional chemical manipulation (Table 2, entries 45 and 46). Compared to pristine polymer P-19a, the partially hydrolyzed polymer P-19a-DP obtained from P-19a through the acidolysis (by subjecting the polymer P-19a to 20×TFA per .sup.tBoc-; TFA as 33% v/v in dichloromethane for 1 hour at room temperature), resulted in improvement of selectivity. Moreover the secondary amines, exposed after acidolysis can be used for additional reactions including, but not limited to dye-labelling, cross-linking and surface coating. Similarly, allyl-functionalities can also be used for post-polymerization transformation that can offer opportunities to incorporate radical and photo-mediated reactions for coatings, crosslinking, and hydrogel formation.

(175) Collectively, these findings demonstrate that these polyionenes have high selectivity and broad spectrum of antimicrobial activity against several microbes over mammalian cells.

(176) TABLE-US-00002 TABLE 2 Minimum inhibitory concentrations (MICs) and hemolytic activity (HC.sub.50) of polymer series, P d-A2 Initial monomer feed ratio Hemo- or A2 of type- B2 lysis, S. monomer 1: type-2 mon- SA EC PA CA HC.sub.50 No. type-1 type-2 monomers omer Code Ref. (ppm) (ppm) (ppm) (ppm) (ppm) 1 1 — — a P-1a-1 6037B 8 8 8 2 >2000 2 1 — — c P-1c-1 6050A 8 8 4 16 >2000 3 3 — — a P-3a 7016  8 16 8 8 >2000 4 3 — — c P-3c 6050C 8 16 16 16 1000 5 5 — — c P-5c-1 6050D 8 16 8 31 >2000 6 6 — — a P-6a 6080A 16 8 8 >500 >2000 7 6 — — b P-6b 6080B 16 8 8 >500 >2000 8 6 — — c P-6c 6080C 16 8 8 >500 >2000 9 4 — — a P-4a 6081A 63 63 63 63 >2000 10 4 — — b P-4b 6081B 31 31 31 16 >2000 11 4 — — c P-4c 6081C 31 31 31 16 >2000 12 1 — — a P-1a-2 6083A 8 8 8 2 >2000 13 1 — — b P-1b 6083B 8 8 8 16 >2000 14 1 — — c P-1c-2 6083C 8 4 4 4 >2000 15 5 — — a P-5a 6084A 8 8 8 63 >2000 16 5 — — b P-5b 6084B 8 8 8 >500 >2000 17 5 — — c P-5c-2 6084C 8 8 8 >500 >2000 18 1 — — a P-1a-3 6104A 8 8 8 2 >2000 19 1 21 1:1 a P-(1:21/1:1)a 6104B 8 8 8 4 >2000 20 1 21 1:9 a P-(1:21/1:9)a 6104C 4 8 8 4 >2000 21 21 — — a  P-21a 6029A 4 4 4 8 >2000 22 1 — — a P-1a-4 7004  8 8 8 2 >2000 23 7 — — a P-7a 7011A 16 8 31 8 >2000 24 7 1 1:1 a P-(7:1/1:1)a 7012A 8 8 16 4 >2000 25 7 1 3:7 a P-(7:1/3:7)a 7020  8 8 8 4 >2000 26 7 1 1:9 a P-(7:1/1:9)a 7012B 8 8 8 4 >2000 27 8 — — a P-8a 6110A 8 8 8 4 >2000 28 8 — — b P-8b 6110B 8 16 8 4 1000-2000 29 8 — — c P-8c 6110C 8 8 8 4 >2000 30 8 21 1:1 a P-(8:21/1:1)a 6110D 2 8 8 4 >2000 31 8 21 1:9 a P-(8:21/1:9)a 6110E 2 8 8 4 >2000 32 9 — — a P-9a 7008  16 16 16 31  63-125 33 10 — — a  P-10a 7007  >500 250 250 125 >8 34 12 — — a  P-12a 7006  >500 31 125 16 >2000 35 13 — — a  P-13a 7014A 8 8 16 8  500-1000 36 14 — — a  P-14a 7050A 4 8 8 250 >1000 37 14 1 1:1 a P-(14:1/1:1)a 7050B 4 4 2 4 >1000 38 14 1 1:3 a P-(14:1/1:3)a 7050C 8 8 8 4 >1000 39 14 21 1:1 a  P-(14:21/1:1)a 7070A 2 8 4 8 >1000 40 14 7 1:1 a P-(14:7/1:1)a 7070C 8 8 16 4 >1000 41 14 7 1:3 a P-(14:7/1:3)a 7070D 8 8 16 8 >1000 42 17 — — a  P-17a 7082B 16 16 8 >500 >1000 43 18 — — a  P-18a 7085A 31 16 31 125 >1000 44 18 1 1:1 a  P-(18:1//1:1)a 7085B 16 8 8 16 >1000 45 19 — — a  P-19a 7014-B 8 16 63 63  500-1000 46 19 — — a P-19a-DP 7014-B-DP 8 8 31 500 >2000 ppm—parts per million; Staphylococcus aureus (SA); Escherichia coli (EC); Pseudomonas aeruginosa (PA); Candida albicans (CA); HC.sub.50: Lowest concentration that produces 50% hemolysis.—means not applicable.

Example 5: Bacterial Mechanism

(177) Based on the MIC and hemolysis data, the polymers with the best activity and selectivity were tested again for bactericidal activity against all microbes. At MIC and 2×MIC, at least a 99.99% reduction in the colony forming units (CFU) was achieved for all the microbes (FIG. 86). There was a 8-14 log reduction in the CFU counts after 18 hours of incubation for all microbes tested at MIC and 2×MIC. This suggested a bactericidal mechanism at a concentration higher than the MIC.

(178) The polymers (P-7a and P-9a) that have shown good antimicrobial properties were chosen to treat microbes using the agar gel assay. At MIC and 2×MIC concentrations, both polymers displayed more than 99.99% killing efficiency against all the microbes tested, suggesting a bactericidal mechanism. As seen from the colony forming units (CFU) counts after 18 hours incubation (FIG. 87), at MIC and 2×MIC, there was an 8 to 16 log reduction in colony counts for all 4 strains of microbes.

Example 6: Killing Kinetics

(179) P-7a, P-(1:7/1:1)a and P-1a-1 were incubated with different strains of microbes at various concentration for 30 seconds before the viability of the microbes was checked using the agar gel assay. For P-7a (FIG. 88), the killing efficiency against PA is >90% at 8×MIC and for 16×MIC, 100% killing efficiency was achieved. A greater than 3 log reduction of SA colonies is seen at 16×MIC after 30 sec incubation. Against EC, the killing efficiency reached 100% at 8×MIC. For CA, only 4×MIC is needed to achieve more than 3 log reduction. With the incorporation of a small amount of amides (P-(1:7/1:1)a), a 30 sec kill (FIG. 89) could be achieved at a lower concentration (8×MIC for all bacteria and 4×MIC for fungi). A 100% kill is observed at 2×MIC against all 4 microbes (FIG. 90) for P-1a-1 showing that more amide groups present lead to better killing efficacy.

Example 7: Synthesis of Gemini-Surfactants (S)

(180) By using excess of functional tertiary amines, under similar conditions to that of polymerization, a library of Gemini-surfactants S were readily accessed. After reaction the surfactant was purified by precipitating thrice into diethylether, to remove DMF and excess tertiary amines, to result in solids. These surfactants are coded by three characters. For, example S-5a, denotes the surfactant synthesized using d-A.sub.2 monomer 5 and B tertiary amine a (FIG. 1 and FIG. 2).

Example 8: Antimicrobial and Hemolytic Activities of Gemini-Surfactants (S)

(181) Similar to polymers, MIC and HC.sub.50 values of the surfactants were evaluated.

(182) The MIC values were highly dependent on both d-A.sub.2 precursor and functional tertiary amine used to synthesize the target surfactant. Comparing different compositions by varying the amine (B; a to h) with a fixed d-A.sub.2 precursor (d-A.sub.2-5), only the derivatives with longer linear alkyl chains (S-5f, S-5g and S-5h with octyl, decyl and dodecyl, respectively), were found to be potent against the pathogens tested (Table 3, entries 1 to 8). Likewise for other d-A.sub.2 precursors (d-A.sub.2-1, d-A.sub.2-3, d-A.sub.2-4 and d-A.sub.2-6), in general, derivatives with decyl and dodecyl derivatives were found to be potent against the pathogens tested (Table 3, entries 9 to 20, S-Xg and S-Xh, where is X is an integer from 1 to 12).

(183) By taking hemolysis into consideration, decyl chains were found to be optimal, offering high potency and relatively good selectivity. Additionally surfactants with decyl chains were evaluated to further probe the role of d-A.sub.2 precursors (Table 3, entries 7, 10, 13, 16, 19 and 21 to 25) and it was found that d-A.sub.2 precursors with hexyl spacers, d-A.sub.2-3 and d-A.sub.2-13, resulting in S-3g and S-13g were optimal.

(184) Collectively these findings demonstrate that these surfactants can be tailored to have broad spectrum of antimicrobial activity against several microbes and tunable selectivity over mammalian cells.

(185) Based on the MIC and hemolysis data, the bactericidal activity of the best candidate (S-3g) was tested against all the microbes. At MIC and 2×MIC, at least a 99.9% reduction in the colony forming units (CFU) was achieved for all the microbes, suggesting a bactericidal mechanism at MIC or higher concentrations (FIG. 91). Rapid killing of pathogens are important for a variety of applications including disinfectants. For this purpose, killing kinetics data was evaluated across multiple strains of microbes. It was found that at 16 and 32×MIC, all four strains of microbes tested were completely killed for 2 minutes (FIG. 92). A higher concentration of 64×MIC was needed to completely eradicate the microbes in 30 seconds.

(186) The ability of an antimicrobial to eradicate microbes in the form of biofilms is important in infection control. S-3g was evaluated against S. aureus and E. coli biofilms. Cell viability and biomass of biofilm for both types of bacteria were found to be reduced in a dose dependent manner (FIG. 93). These findings demonstrate that these small molecule Gemini-surfactants are promising for use in consumer-care applications as potent disinfectant and/or preservatives.

(187) TABLE-US-00003 TABLE 3 Minimum inhibitory concentrations (MICs) and hemolytic activity (HC.sub.50) of Gemini-surfactant series, S S. SA EC PA CA Hemolysis, No. Code Ref. (ppm) (ppm) (ppm) (ppm) HC.sub.50(ppm) 1 S-5a 6044A >500 >500 >500 >500 >2000 2 S-5b 6044D >500 >500 >500 >500 >2000 3 S-5c 6044C >500 >500 >500 >500 >2000 4 S-5d 6039A >500 >500 >500 >500 >2000 5 S-5e 6044B >500 >500 >500 >500 >2000 6 S-5f 6039B 31 250 >500 125 >2000 7 S-5g 6092D 2 4 63 2 125-250 8 S-5h 6039C 2 4 16 1 16-31 9 S-1f 6045E 31 250 >500 125 >2000 10 S-1g 6092A 1 2 16 2 125-250 11 S-1h 6045F 16 8 31 2 125-250 12 S-3f 6048E 4 16 500 8  500-1000 13 S-3g 6092B 1 1 8 2 125-250 14 S-3h 6048F 4 16 16 2 125-250 15 S-4f 6071E 2 8 250 125 250-500 16 S-4g 6092C 1 1 8 1  63-125 17 S-4h 6071F 8 8 31 2  63-125 18 S-6f 6073E 16 125 >500 16 >2000 19 S-6g 6092E 2 4 63 4 250-500 20 S-6h 6073F 1 2 8 4 31-63 21 S-9g 7015B 1 2 8 1 125-250 22  S-13g 7015C 1 1 4 1  63-125 23 S-7g 7019D 1 1 8 1  63-125 24  S-10g 7019B 16 500 500 16  63-125 25  S-12g 7019A 1 2 8 2 125-250

Example 9: Surgical Scrub Formulation

(188) Several formulations were prepared and tested for their antimicrobial efficacy for use as a surgical scrub. Two different hydroxypropyl methyl cellulose (HPMC) (H3785 and H7509) and Igepal (520 and 630) were used and the outcome of the different combinations was tabulated in Table 4. The compositions were made to 100% with deionized water. 1 wt % of HPMC was chosen in the preferred composition as the mixture is homogeneous. The most preferred formulation is shown in Table 5 and was chosen due to homogeneity, transparency and fluidity among all the different combinations from Table 4.

(189) TABLE-US-00004 TABLE 4 Outcome of different compositions of HPMC and Igepal. HPMC (wt %) Igepal (wt %) H3785 H7509 520 630 Remarks — 1 — 5 Single phase, slightly opaque, slightly viscous — 2 — 5 2 phase, slightly opaque, viscous — 1 5 — Single phase, clear, slightly viscous — 2 5 — 2 phase, slightly opaque, viscous 1 — — 5 Single phase, clear, fluid 2 — — 5 2 phase, slightly opaque, slightly viscous 1 — 5 — 2 phase, slightly opaque, slightly viscous 2 — 5 — 2 phase, slightly opaque, slightly viscous

(190) TABLE-US-00005 TABLE 5 Optimized composition. P-1a-1 (6037B) 0.5 wt %   HPMC 3785 1 wt % Igepal 630 5 wt % N,N-Bis(2-hydroxyethyl)dodecanamide (HDA) or 5 wt % N-Decyl-b-D-glucopyranoside (DGP)

(191) After incubating the bacteria with the formulation at different time points (10 seconds, 30 seconds and 2 minutes), the mixture was plated onto agar plates. It was found that regardless of the initial colony forming units (CFU) of bacteria (3×10.sup.5 or 3×10.sub.8 CFU/mL) used, no CFU was found on the agar plate (Table 6). This finding demonstrated that the formulation can be used as a hand wash or surgical scrub.

(192) TABLE-US-00006 TABLE 6 Killing effectiveness of different formulations with different initial bacteria loading and various incubation time periods. Bacteria Incubation SA EC PA Count Time HDA DGP HDA DGP HDA DGP 3 × 10.sup.5 2 minutes 100% 100% 100% 100% 100% 100% CFU/mL 30 seconds 100% 100% 100% 100% 100% 100% 3 × 10.sup.8 2 minutes 100% 100% 100% 100% 100% 100% CFU/mL 30 seconds 100% 100% 100% 100% 100% 100% 10 seconds 100% 100% 100% 100% 100% 100%

Example 10: In Vivo Toxicity

(193) The LD.sub.50 and LD.sub.5 values of P-1a-1 (6037B) were determined to be 67.5 and 37.3 mg/kg (mouse weight) by intraperitoneal injection, respectively. The ED.sub.50/ED.sub.95 values of P-1a-1 (6037B) and imipenem were determined to be 0.62/3.08 mg/kg and 2.75/20.0 mg/kg, respectively, indicating that the polymer was more effective than the antibiotic imipenem. At doses of 1.0 mg/kg and 2.0 mg/kg for P-1a-1 (6037B) and imipenem respectively, P-1a-1 (6037B) saved more infected mice than imipenem (survival: 80% and 60% for 6037B and imipenem, respectively) (FIG. 94). In addition, the treatment with P-1a-1 (6037B) and imipenem cleared the bacteria from blood and the major organs, with P-1a-1 (6037B) being significantly more effective, as shown in FIG. 95. In particular, more than 99.9% bacteria were removed after P-1a-1 (6037B) treatment.

INDUSTRIAL APPLICABILITY

(194) The compounds and compositions as defined above may have useful applications as an antimicrobial agent in personal care goods such as cosmetics, deodorant and soap, as well as in the medical field, such as in hand washes and surgical scrubs. The compounds as defined above may also have useful applications in treating antimicrobial infections. The compounds and compositions as defined above may be ideal for use in personal care and biomedical applications.

(195) The compounds and compositions as defined above may also have useful applications as food additives or preservatives, coatings, precursors for cross-linked resins, ingredients for additive manufacturing, surfactants, and agents to selectively promote a certain microbial strain by killing other agents, for instance, by promoting fungi, while killing bacterial strains.

(196) It will be apparent that various other modifications and adaptations of the invention will be apparent to the person skilled in the art after reading the foregoing disclosure without departing from the spirit and scope of the invention and it is intended that all such modifications and adaptations come within the scope of the appended claims.