Granted Patent B2

US 12,551,553 · App. 17/296,689 · Granted Feb 17, 2026

Inventors: Pol Harvengt (Rixensart, BE); Philippe Jehoulot (Rixensart, BE); Loic Le Gourrierec (Rixensart, BE); Demostene Sifakakis (Rixensart, BE); Laurent Strodiot (Rixensart, BE)

Assignee: GlaxoSmithKline Biologicals SA

A61K39/39A61K47/24A61K47/28A61P37/04A61K2039/55555A61K2039/55572A61K2039/55577Y02A50/30

Abstract

Claims (73)

1 . A method of manufacturing a liposomal adjuvant using a microfluidic device; the liposomal adjuvant comprising liposomes; the liposomes comprising: (i) a phosphatidylcholine lipid and (ii) an aminoalkyl glucosaminide phosphate compound (AGP) or a glucopyranosyl lipid adjuvant (GLA); the microfluidic device comprising a mixing chamber, a first inlet, at least two second inlets, and an outlet; the first inlet and the at least two second inlets being in fluid communication with the mixing chamber; the mixing chamber being in fluid communication with the outlet; each of the first inlet and the at least two second inlets having a cross-sectional area of 0.16 mm 2 or less; the method comprising:

(a) mixing in the mixing chamber a first solution and a second solution; the first solution comprising: (i) a solvent, (ii) the AGP or the GLA and (iii) 100-170 mg of the phosphatidylcholine lipid; the solvent comprising 70-90% v/v ethanol and 10-30% v/v isopropyl alcohol: the second solution comprising water; the first solution being delivered to the mixing chamber by the first inlet; and the second solution being delivered to the mixing chamber by the at least two second inlets; thereby obtaining the liposomal adjuvant in the mixing chamber; the liposomal adjuvant flowing through the outlet; and

(b) removing the solvent from the liposomal adjuvant.

2 . The method of claim 1 further comprising adding a saponin between (a) and (b).

3 . The method of claim 1 further comprising: (c) adding a saponin.

4 . The method of claim 1 , wherein the second solution further comprises a saponin and the liposomal adjuvant further comprises the saponin.

5 . The method of claim 4 , wherein the saponin comprises QS-21.

6 . The method of claim 5 , wherein the liposome comprises the GLA and the GLA is:

or a salt thereof.

7 . The method of claim 6 , wherein the phosphatidylcholine lipid is DOPC.

8 . The method of claim 7 , wherein the liposomes have a Z-average diameter of from 95 nm to 120 nm and a polydispersity of 0.3 or lower.

9 . The method of claim 5 , wherein the liposome comprises the GLA and the GLA is:

or a salt thereof.

10 . The method of claim 9 , wherein the phosphatidylcholine lipid is DOPC.

11 . The method of claim 10 , wherein the liposomes have a Z-average diameter of from 95 nm to 120 nm and a polydispersity of 0.3 or lower.

12 . The method of claim 1 , wherein the phosphatidylcholine lipid is dioleoyl phosphatidylcholine (DOPC).

13 . The method of claim 1 , wherein the liposomes have a Z-average diameter of from 95 nm to 120 nm and a polydispersity of 0.3 or lower.

14 . The method of claim 1 further comprising mixing the liposomal adjuvant with an immunogen or a polynucleotide encoding the immunogen, thereby obtaining an adjuvanted immunogenic composition.

15 . The method of claim 1 ; the liposomes further comprising cholesterol; the solvent being divided into a first portion and a second portion; the method further comprising before (a):

(i) preparing a suspension of the AGP or the GLA in the first portion of the solvent, thereby obtaining a suspension;

(ii) combining the suspension with the phosphatidylcholine lipid and the cholesterol, thereby obtaining a first intermediate;

(iii) adding the second portion of the solvent to the first intermediate, thereby obtaining a second intermediate;

(iv) mixing, thereby obtaining the first solution.

16 . The method of claim 1 ; the liposomes further comprising cholesterol; the solvent being divided into a first portion, a second portion, and a third portion, the method further comprising before (a):

i) preparing a suspension of the AGP or the GLA in the first portion of the solvent, thereby obtaining a suspension;

(ii) combining the suspension with the phosphatidylcholine lipid and the cholesterol, thereby obtaining a first intermediate;

(iii) adding the second portion of the solvent to the first intermediate, thereby obtaining a second intermediate;

(iv) mixing the second intermediate, thereby obtaining a third intermediate;

(v) adding the third portion of the solvent to the third intermediate, thereby (v) obtaining the first solution.

17 . The method of claim 1 , wherein the liposomes comprise the AGP and the AGP has the following structure:

wherein:

m is 0 to 6;

n is 0 to 4;

X is O or S;

Y is O or NH;

Z is O or H;

each of R 1 , R 2 , and R 3 is selected independently from the group consisting of a C 1-20 acyl and a C 1-20 alkyl;

R 4 is H or methyl;

R 5 is selected from the group consisting of —H, —OH, —(C 1 -C 4 )alkoxy, —PO 3 R 8 R 9 , —OPO 3 R 8 R 9 , —SO 3 R 8 , —OSO 3 R 8 , —NR 8 R 9 , —SR 8 , —CN, —NO 2 , —CHO, —CO 2 R 8 , and —CONR 8 R 9 ,

wherein R 8 and R 9 are each independently selected from H and (C 1 -C 4 ) alkyl; and

each of R 6 and R 7 is independently H or PO 3 H 2 ;

or a pharmaceutically acceptable salt thereof.

18 . The method of claim 1 , wherein the liposomes comprise the AGP and AGP has the following structure:

wherein:

X is O or S;

Y is O or NH;

Z is O or H;

each of R 1 , R 2 , and R 3 is selected independently from the group consisting of a C 1-20 acyl and a C 1-20 alkyl;

and R 4 is H or methyl;

or a pharmaceutically acceptable salt thereof.

19 . The method according to claim 1 , wherein the liposomes comprise the AGP and the AGP is:

or a salt thereof.

20 . The method of claim 1 , wherein the liposome comprises the GLA and the GLA is:

wherein:

R 1 , R 3 , R 5 , and R 6 are each independently C 11-20 alkyl; and

R 2 and R 4 are each independently C 12-20 alkyl; or

salts thereof.

21 . The method of claim 1 , wherein the liposomes comprise the GLA and the GLA is:

wherein:

R 1 , R 3 , and R 6 are each independently C 11-20 alkyl; and

R 2 and R 4 are each independently C 12-20 alkyl; or

a salt thereof.

22 . The method of claim 1 , wherein the liposomes comprise the GLA and the GLA is:

wherein:

R 1 , R 3 and R 6 are each independently C 11-20 alkyl; and

R 2 and R 4 are each independently C 12-20 alkyl; or

a salt thereof.

23 . The method of claim 1 , wherein the GLA is:

or a salt thereof.

24 . The method of claim 1 , wherein the liposomes comprise the GLA and the GLA is:

or a salt thereof.

25 . The method of claim 1 , wherein the liposomes further comprise a sterol and the first solution further comprises the sterol.

26 . A method of manufacturing a liposomal concentrate of use in the preparation of a liposomal adjuvant using a microfluidic device; the liposomal concentrate comprising liposomes; the liposomes comprising: (i) a phosphatidylcholine lipid and (ii) an AGP or a GLA; the microfluidic device comprising a mixing chamber, a first inlet, at least two second inlets, and an outlet; the first inlet and the at least two second inlets being in fluid communication with the mixing chamber; the mixing chamber being in fluid communication with the outlet; each of the first inlet and the at least two second inlets having a cross-sectional area of 0.16 mm 2 or less; the method comprising mixing in the mixing chamber a first solution and a second solution, the first solution comprising: (i) a solvent, (ii) the AGP or the GLA, and (iii) 100-170 mg of the phosphatidylcholine lipid; the solvent comprising 70-90% v/v ethanol and 10-30% v/v isopropyl alcohol; the second solution comprising water; the first solution being delivered to the mixing chamber by the first inlet; and the second solution being delivered to the mixing chamber by the at least two second inlets; thereby obtaining the liposomal adjuvant in the mixing chamber; and the liposomal adjuvant flowing through the outlet.

Assignments (1)

Priority Claims (1)

Continuity (1)

References Cited (164)