Methods of making liposomes containing hydro-monobenzoporphyrin photosensitizer

Claim 3

The method of claim 1, wherein said molar ratio of hydro-monobenzoporphyrin photosensitizer to phospholipids is about 1:8.0 or more phospholipid.

Claim 4

The method of claim 1, wherein said organic solvent is methylene chloride.

Claim 5

The method of claim 1, wherein said organic solvent is removed in step "b." by evaporation under reduced pressure.

Claim 6

The method of claim 1, wherein said aqueous solution comprises a disaccharide or polysaccharide.

Claim 7

The method of claim 4, wherein said disaccharide or polysaccharide is selected from lactose or trehalose.

Claim 8

The method of claim 4, wherein the concentration ratio of disaccharide or polysaccharide to said mixture of phospholipids is about 10-20 to 0.5-6.0.

Claim 9

The method of claim 1, wherein said hydro-monobenzoporphyrin (Gp) photosensitizer has one of the formulae 1-6 set forth in FIG. 1 and has one or more light absorption maxima between 670-780 nm, or is a metalated or labeled form thereof, wherein each R 1 and R 2 is independently selected from the group consisting of carbalkoxy (2-6C), alkyl (1-6C) sulfonyl, aryl (6-10C) sulfonyl, aryl (6-10C), cyano, and --CONR 5 CO-- wherein R 5 is aryl (6-10C) or alkyl (1-6C); each R 3 is independently carboxyalkyl (2-6C) or a salt, amide, ester or acylhydrazone thereof, or is alkyl (1-6C); and R 4 is --CHCH 2 ; --CHOR 4' wherein R 4' is H or alkyl (1-6C), optionally substituted with a hydrophilic substituent; --CHO; --COOR 4' ; --CH(OR 4' )CH 3 ; --CH(OR 4' )CH 2 OR 4' ; --CH(SR 4' )CH 3 ; --CH(NR 4' 2 )CH 3 ; --CH(CN)CH 3 ; --CH(COOR 4' )CH 3 ; --CH(OOCR 4' )CH 3 ; --CH(halo)CH 3 ; --CH(halo)CH 2 (halo); an organic group of less than 12C resulting from direct or indirect derivatization of a vinyl group; or R 4 is a 1-3 tetrapyrrole-type nucleus of the formula --L--P, wherein --L-- is selected from the group consisting of ##STR3## and P is a second Gp, which is one of the formulae 1-6 but lacks R 4 and is conjugated to L through the position shown as occupied by R 4 , or another porphyrin group.

Claim 10

The method of claim 8 wherein, when P is said another porphyrin group, P has the formula: ##STR4## wherein: each R is independently H or lower alkyl (1-4C); two of the four bonds shown as unoccupied on adjacent rings are joined to R 3 ; one of the remaining bonds shown as unoccupied is joined to R 4 ; and the other is joined to L; with the proviso that, if R 4 is CHCH 2 , said R 3 groups cannot both be carbalkoxyethyl.

Claim 11

The method of claim 8, wherein each R 3 is --CH 2 CH 2 COOH or salt, amide, ester or acylhydrazone thereof.

Claim 12

The method of claim 8, wherein each of R 1 and R 2 is carbalkoxy (2-6C).

Claim 13

The method of claim 1, wherein the photosensitizer is BPD-MA having the formula of FIG. 2.

Claim 14

The method of claim 1, wherein said hydrating step "c." is accomplished at a temperature at or below room temperature.

Claim 15

The method of claim 1, wherein said homogenizing or reducing step "d." is accomplished at a temperature at or below room temperature.

Claim 16

The method of claim 1, wherein said particle size is reduced to a range of below about 250 nm.

Claim 17

The method of claim 15 wherein the particle size is reduced to below about 220 nm.

Claim 18

A pharmaceutical composition containing liposomes in the particle size range of about 150 to 300 nm, wherein said liposomes comprise: a. a therapeutically acceptable amount of a photosensitizer and b. a mixture of phospholipids comprising: (1) egg phosphatidyl glycerol ("EPG") and (2) dimyristoyl phosphatidyl choline ("DMPC"), wherein the molar ratio of said photosensitizer and said mixture of phospholipids is about 1:7.0 or more phospholipid.

Claim 19

A pharmaceutical composition containing liposomes in the particle size range of about 150 to 300 nm, wherein said liposomes comprise: a. a therapeutically acceptable amount of a photosensitizer and b. a mixture of phospholipids comprising: (1) egg phosphatidyl glycerol ("EPG") and (2) dimyristoyl phosphatidyl choline ("DMPC"), wherein the molar ratio of said photosensitizer and said mixture of phospholipids is about 1:7.0 or more phospholipid.

Claim 20

The composition of claim 17, wherein said molar ratio of photosensitizer to said mixture of phospholipids is about 1:8.0 or more phospholipid.

Claim 21

The composition of claim 17, wherein said hydro-monobenzoporphyrin (Gp) has one of the formulae 1-6 set forth in FIG. 1 and has one or more light absorption maxima between 670-780 nm, or is a metalated or labeled form thereof, wherein: each R 1 and R 2 is independently selected from the group consisting of carbalkoxy (2-6C), alkyl (1-6C) sulfonyl, aryl (6-10C) sulfonyl, aryl (6-10C), cyano, and --CONR 5 CO-- wherein R 5 is aryl (6-10C) or alkyl (1-6C); each R 3 is independently carboxyalkyl (2-6C) or a salt, amide, ester or acylhydrazone thereof, or is alkyl (1-6C); and R 4 is --CHCH 2 ; --CHOR 4' wherein R 4' is H or alkyl (1-6C), optionally substituted with a hydrophilic substituent; --CHO; --COOR 4' ; --CH(OR 4' 2 )CH 3 ; --CH(OR 4' )CH 2 OR 4' ; --CH(SR 4' )CH 3 ; --CH(NR 4' )CH 3 ; --CH(CN)CH 3 ; --CH(COOR 4' )CH 3 ; --CH(OOCR 4' )CH 3 ; --CH(halo)CH 3 ; --CH(halo)CH 2 (halo); an organic group of less than 12C resulting from direct or indirect derivatization of a vinyl group; or R 4 is a 1-3 tetrapyrrole-type nucleus of the formula --L--P, wherein --L-- is selected from the group consisting of ##STR5## and P is a second Gp, which is one of the formulae 1-6 but lacks R 4 and is conjugated to L through the position shown as occupied by R 4 , or another porphyrin group.

Claim 22

The composition of claim 19 wherein, when P is said another porphyrin group, it has the formula: ##STR6## wherein each R is independently H or lower alkyl (1-4C); two of the four bonds shown as unoccupied on adjacent rings are joined to R 3 ; one of the remaining bonds shown as unoccupied is joined to R 4 ; and the other is joined to L; with the proviso that, if R 4 is CHCH 2 , said R 3 groups cannot both be carbalkoxyethyl.

Claim 23

The composition of claim 20, wherein each R 3 is --CH 2 CH 2 COOH or a salt, amide, ester or acylhydrazone thereof.

Claim 24

The composition of claim 19, wherein each of R 1 and R 2 is carbalkoxy (2-6C).

Claim 25

The composition of claim 19, wherein the hydro-monobenzoporphyrin photosensitizer is BPD-MA having the formula of FIG. 2.

Claim 26

The composition of claim 19, wherein said particle size is below about 220 nm.