Substance-containing vesicle, and production method therefor

Patent metadata

Field	Value
Publication number	US-9782358-B2
Application number	US-201514839854-A
Country	US
Kind code	B2
Filing date	Aug 28, 2015
Priority date	Mar 1, 2013
Publication date	Oct 10, 2017
Grant date	Oct 10, 2017

Abstract

Provided is a monodisperse agglomerate of a substance-containing vesicle filled with a substance at a concentration higher than conventionally possible. A mixed solution, in which a target substance is included in an aqueous medium, is mixed with a monodisperse agglomerate of a crosslinked vesicle comprising a prescribed polymer which includes a first polymer, i.e. a block copolymer having uncharged hydrophilic segments and first charged segments, and a second polymer having second charged segments carrying a charge opposite to that of the first charged segments, and in which the first polymer and/or the second polymer are/is crosslinked. As a result, the crosslinked vesicle is made to contain the target substance.

First claim

What is claimed is:

1. A monodisperse population of substance-encapsulating crosslinked vesicles, each of the substance-encapsulating crosslinked vesicles comprising:
a crosslinked membrane comprising a first polymer, which is block copolymer having an uncharged hydrophilic segment and a first charged segment, and a second polymer, which has a second charged segment having a charge opposite to the charge of the first charged segment, the first and/or the second polymer(s) being crosslinked;
an inner aqueous phase surrounded by the crosslinked membrane; and
a target substance encapsulated in the inner aqueous phase,
wherein the concentration of the target substance encapsulated in the inner aqueous phase is higher than 5mg/mL.

2. The monodisperse population of substance-encapsulating crosslinked vesicles according to claim 1 , having a polydispersity index of 0.2 or lower.

3. The monodisperse population of substance-encapsulating crosslinked vesicles according to claim 1 or 2 , wherein the weight-average molecular weight of the target substance is between 10000 and 40000.

4. The monodisperse population of substance-encapsulating crosslinked vesicles according to claim 2 , wherein the first and/or the second polymer(s) is(are) crosslinked with one or more crosslinking bonds selected from the group consisting of a crosslinking bond formed between cationic groups, a crosslinking bond formed between anionic groups, and a crosslinking bond formed between a cationic group and an anionic group, and the molar ratio of the cationic groups and/or anionic groups forming crosslinking bonds to the cationic groups and/or anionic groups contained in the crosslinked membrane is 35% or more.

5. A substance-encapsulating crosslinked vesicles comprising:
a crosslinked membrane comprising a first polymer, which is block copolymer having an uncharged hydrophilic segment and a first charged segment, and a second polymer, which has a second charged segment having a charge opposite to the charge of the first charged segment, the first and/or the second polymer(s) being crosslinked;
an inner aqueous phase surrounded by the crosslinked membrane; and
a first target substance and a second target substance encapsulated in the inner aqueous phase, wherein the first target substance having a smaller molecular weight than the molecular weight of the second target substance, and the second target substance is a crowding agent,
wherein the first target substance is more stable than when the first target substance is contained in the inner aqueous phase in the absence of the second target substance.

6. A method of producing a substance-encapsulating vesicle, comprising mixing a monodisperse population of vacant crosslinked vesicles with a mixture liquid containing the target substance in an aqueous medium to form a monodisperse population of substance-encapsulating crosslinked vesicles,
wherein each of the vacant crosslinked vesicles comprises: a crosslinked membrane comprising a first polymer, which is block copolymer having an uncharged hydrophilic segment and a first charged segment, and a second polymer, which has a second charged segment having a charge opposite to the charge of the first charged segment, the first and/or the second polymer(s) being crosslinked; and an inner aqueous phase surrounded by the crosslinked membrane, wherein the inner aqueous phase does not contain the target substance, and
wherein each of the substance-encapsulating crosslinked vesicles comprises: the crosslinked membrane comprising the first polymer and the second polymer, the first and/or the second polymer(s) being crosslinked; and the inner aqueous phase surrounded by the crosslinked membrane, wherein the inner aqueous phase contains the target substance.

7. The method according to claim 6 , wherein the monodisperse population of vacant crosslinked vesicles, the monodisperse population of substance-encapsulating crosslinked vesicles, the monodisperse population of vacant non-crosslinked vesicles, and the monodisperse population of substance-encapsulating non-crosslinked vesicles each have a polydispersity index of 0.2 or less.

8. The method according to claim 6 or 7 , wherein the weight-average molecular weight of the target substance is between 10000 and 40000, and the concentration of the target substance in the mixture liquid is more than 5 mg/mL.

9. The method according to claim 8 , wherein both in the vacant crosslinked vesicles and the substance-encapsulating crosslinked vesicles, the first and/or the second polymer(s) is(are) crosslinked with one or more crosslinking bonds selected from the group consisting of a crosslinking bond formed between cationic groups, a crosslinking bond formed between anionic groups, and a crosslinking bond formed between a cationic group and an anionic group, and the molar ratio of the cationic groups and/or anionic groups forming crosslinking bonds to the cationic groups and/or anionic groups contained in the crosslinked membrane is 35% or more.

10. The method according to claim 6 , further comprising: reacting the monodisperse population of substance-encapsulating crosslinked vesicles with a crosslinker which can react with the first and/or the second polymer(s).

11. A method of producing a substance-encapsulating vesicle, comprising:
mixing a first substance-encapsulating crosslinked vesicle, which encapsulates a first target substance, with a mixture liquid containing a second target substance in an aqueous medium to form a second substance-encapsulating crosslinked vesicle,
wherein the first substance-encapsulating crosslinked vesicle comprises: a crosslinked membrane comprising a first polymer, which is block copolymer having an uncharged hydrophilic segment and a first charged segment, and a second polymer, which has a second charged segment having a charge opposite to the charge of the first charged segment, the first and/or the second polymer(s) being crosslinked; and an inner aqueous phase surrounded by the crosslinked membrane, wherein the inner aqueous phase contains the first target substance, and
wherein the second substance-encapsulating crosslinked vesicle comprises: the crosslinked membrane comprising the first polymer and the second polymer, the first and/or the second polymer(s) being crosslinked; and the inner aqueous phase surrounded by the crosslinked membrane, wherein the inner aqueous phase contains the first target substance and the second target substances.

12. The method according to claim 11 , wherein a monodisperse population of the first substance-encapsulating crosslinked vesicles is used to thereby produce a monodisperse population of the second substance-encapsulating crosslinked vesicles.

13. The method according to claim 12 , wherein the concentration of the second target substance contained in the mixture liquid is high enough to prevent formation of a monodisperse population of substance-encapsulating non-crosslinked vesicles each encapsulating both the first and second target substances when a monodisperse population of first non-crosslinked vesicles is mixed with the mixture liquid, wherein the monodisperse population of first substance-encapsulating non-crosslinked vesicles differs from the monodisperse population of first substance-encapsulating crosslinked vesicles only in that the first and/or the second polymer(s) is(are) not crosslinked.

14. The method according to any one of claims 11 to 13 , further comprising
forming the first substance-encapsulating crosslinked vesicle by contacting a vacant crosslinked vesicle a mixture liquid containing the first target substance in an aqueous medium, wherein the

Assignees

• Japan Science & Tech Agency

Inventors

• Kataoka Kazunori
• Kishimura Akihiro
• Anraku Yasutaka
• Goto Akinori

Classifications

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title

• A61P35/00

  Antineoplastic agents · CPC title

• A61K9/5146 Primary

  obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, polyamines, polyanhydrides · CPC title

• C08G69/40

  Polyamides containing oxygen in the form of ether groups (C08G69/12, C08G69/32 take precedence) · CPC title

• C08L77/04

  Polyamides derived from alpha-amino carboxylic acids (C08L77/10 takes precedence) · CPC title